ColorViewI en

Manual
ColorView I
User Manual
ColorViewI
3.2 MegaPixel CCD camera
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ColorView I
The ColorView I - camera 4

ColorView I
Acquiring images 5
WHEN CAN COLORS APPEAR TO BE FALSE? .....................6

WHEN CAN THE IMAGE BE TOO DARK? ..............................7
CALIBRATING INTELLIGENT EXPOSURE .............................9
EXPOSURE TIME LOCK ........................................................10
Acquiring images with manual controls 11
FURTHER OPTIONS IN THE CAMERA CONTROL ..............17
Online shading correction 23
USING THE SHADING CORRECTION ..................................24

HOW THE SHADING CORRECTION FUNCTIONS ...............24
ACQUIRING REFERENCE IMAGES ......................................24
USE OF THE ONLINE SHADING CORRECTION ..................29
TECHNICAL NOTE .................................................................29
Using the black & white mode 30

ECVI0807
ACQUISITIONS WITH REAL TIME FFT .................................31

ColorView I
NetCam 34
INSTALLING THE WEB SERVER .......................................... 35

NETCAM IN THE INTRANET ................................................. 35
NETCAM IN THE INTERNET ................................................. 36
USING NETCAM ..................................................................... 37
Setting up the camera 39
LOGICAL INPUT CHANNELS ................................................ 39

INPUT TAB ............................................................................. 40
DISPLAY TAB ......................................................................... 43
FORMAT TAB ......................................................................... 47
Appendix I: Microscope settings 49
ILLUMINATION ....................................................................... 49
OBJECTIVE, TOTAL MAGNIFICATION AND USEFUL MAGNI-
FICATION ............................................................................... 51
Appendix II: How to install the camera 52
INSTALLATION PROCEDURE - WINDOWS 2000 ................ 54

INSTALLATION PROCEDURE - WINDOWS XP ................... 66
ColorView III
In case of problems with the camera... 77
WEEE DECLARATION 78
CE DECLARATION OF CONFORMITY 79
ColorView I • The ColorView I - camera
ColorView I
The ColorView I - camera
What is the ColorView I is a quick, high resolution CCD color camera used for biomed-
ColorView I? ical and material sciences applications. It was especially developed for
acquisitions using light microscopy, has a special compact design, and can
be connected to all types of light-microscopes via the C-mount. Its high
dynamic scope enables it to acquire images which have very light and very
dark areas
Technical data ColorView I
Dynamic range 3 x 12 bit using color mode
12 bit using black & white mode
4
Exposure times 100 µs to 160 s

Frame rates 29,5 / s @ Binning 3
11,1/ s @ Binning 2
35,6/ s with full resolution
Resolution options 2080 x 1544,
1040 x 772,
688 x 414 in color mode
1040 x 772 in black & white mode
Camera Mount Standard C-Mount
PC-interface Firewire (IEEE 1394)
System Requirements To properly use ColorView I, the following system requirements for your
computer and your image analysis system should be taken into consider-
ation:
Hard- / Software Required
CPU P4 with at least 1,8 GHz
Memory 512 MB
Hard Disk Storage 30 GB
Interface Firewire (IEEE 1394)
Overview of the chapters
Hard- / Software Required

Operating system Windows 2000 Professional,
Windows XP Pro
Image analysis program analySIS FIVE, build 1054 or higher
cell^A, cell^B, cell^D, cell^F, cell^P
Modern cameras with the respective software offer numerous possibilities
for correcting and modifying the acquired image. They enable the image to
be optimized quickly. However, the entire system, the sample, the micro-
scope with all its possibilities, and the camera all must be optimally set in
order to attain the best possible results.
Overview of the chapters

"Acquiring images" describes the automated acquisition of images with the
help of the Intelligent Exposure function. In the majority of situations you will
be able to use this method to acquire high quality images.
Should you not attain the required acquisition quality with Intelligent Expo-
sure, or should you basically prefer to work with manual controls, "Acquiring
images with manual controls" describes how to work with this method.
ColorView I offers you an online shading correction with which you can
correct images that have not been uniformly illuminated. Chapter "Online
shading correction" describes the setting up and use of this function.
Chapter "Using the black & white mode" describes the possibilities your
camera offers you for making monochrome acquisitions.
Chapter "NetCam" describes the use of the NetCam add-in for making direct
transmissions of images via the Internet or an intranet.
In chapter "Setting up the camera" you can learn how to change the
camera's settings, and how you can define different input channels for
different tasks.
"Appendix I: Microscope settings" provides you with a short overview of how
you select the correct settings for your microscope in order to achieve the
best results.
5
"Appendix II: How to install the camera" contains detailed instructions on

how to install the camera under the Windows 2000 and the Windows XP
operating systems.
Acquiring images
Your image analysis program comes with a procedure for automatically
acquiring images. By using Intelligent Exposure you can acquire high quality
images with just a few mouse clicks.
1) Click the Intelligent Exposure button in the Standard button bar.
" Should you use Intelligent Exposure for the first time with this
camera, you will be asked to perform a calibration (see Calibrating
Intelligent Exposure, p. 9).
" The Intelligent Exposure dialog box will be opened.
ColorView I • When can colors appear to be false?
2) Click the Acquire > button.

" The complete acquired image will be shown in the document
window. Check if the desired image segment is shown in this
setting.
" The exposure times for the live-image and for a snapshot are
displayed in the dialog box's status bar.
3) Find the place on the sample that interests you.
4) Clicking the Focus mode button will enable you to bring the image into
focus.
" The central image segment will then be magnified to the point
where one camera pixel is equivalent to one pixel on the image.
This setting enables you to focus optimally.
5) Bring the image into focus.
" To help you do this, you can use the sharpness monitor.
6) Click the Focus mode button to return to the complete image.
7) Then click the Snapshot button to acquire a concluding image with the
settings you have chosen.
" This image will remain available to you even when the Intelligent
6
Exposure dialog box has been closed, while the live-image will be
deleted.
When can colors appear to be false?

For the color reproduction, the correct settings for the camera's three color
channels, red, green and blue, are decisive.
When is it necessary to perform a white balance?

The most important criterion for the color settings of the camera is the
comparison between the image in the eyepiece and its display on your
monitor. A white balance can be made necessary by the following changes:
• The lamp is exchanged
• The lamp voltage is changed.
Related Topics
Sharpness Monitor 20
When can the image be too dark?
This is how you perform a white balance with Intelligent Expo-

sure
1) Procure a sample that is suitable for a white balance.
" For reflected light microscopy this will be a standardized gray card.
Should one not be available, you can also use white laboratory
porcelain or neutral ND-filter paper.
" When using the transmitted light method, look for a position through
which the light passes through the cover slip, embedding material,
and microscope slide, but not through the specimen. Should such
a position not be present, remove the sample and perform the white
balance without a sample.
2) Position the sample under your microscope and start Intelligent Expo-
sure.
3) Click the White balance button to set an ROI (Region of Interest) for the
white balance with Intelligent Exposure, and to activate the white
balance.
" Pull the ROI's frame shown in the image document to the desired
position and set its size by moving the mouse and keeping the right
mouse button depressed.
" Rightclick to confirm position and size of the ROI.
4) Then continue as usual.
The white balance will then remain activated until you once more click the
White balance button.
When can the image be too dark?

Very bright image segments or ones that reflect strongly, lead to a false
exposure time.
Intelligent Exposure calculated the exposure time that is necessary by
analyzing the signals. Normally, when this is done, all of the pixels will be
used. Very bright areas in an image, or areas that reflect strongly, cause the
exposure time to be reduced, to also enable these areas to be correctly
7
displayed. Areas that are not so bright, or don't reflect so strongly, will then
appear to be darker than they are.
This is how you avoid your images being made dark by bright
areas
In this example a graphite sample will be acquired.
1) Start Intelligent Exposure and click the Snapshot button.
" The darker area of the sample will then be displayed very darkly.
ColorView I • When can the image be too dark?
In the default setting,

scratches and inclusions
don't stand out very much
from the background.
This makes an automatic
evaluation difficult.
2) Click the Set ROI for White Balance button.

" The mouse pointer appears in the image. On its tip, a red rectangle,
the so-called ROI (Region of Interest) can be seen.
3) Move the mouse to alter the position of the ROI. Change the size of the
ROI by keeping the mouse key depressed and moving the mouse.
4) Position the ROI in an image segment that you can display well.
" While you are moving the ROI, the exposure time will not yet be
changed.
5) Click your right mouse button to fix the size and position of the ROI.
" Intelligent Exposure will then readjust the exposure time to have the
area within the ROI optimally displayed.
When the ROI for the calcu-
lation of the exposure time is
moved to the darker area of
the sample, the sample and
its structures will be brighter,
and more clearly, displayed.
With these settings, an auto-
matic evaluation is made
much easier.
8
This is how you avoid your images being made dark by strongly
reflecting areas
In this example, a coin with strongly reflecting points will be acquired.
1) Start Intelligent Exposure and click the Snapshot button.
" The sample will be displayed very darkly, since the reflecting points
simulate a bright sample area when the exposure time is calcu-
lated.
Calibrating Intelligent Exposure
With the default setting of

the exposure time correc-
tion, the reflecting areas on
the surface of this copper
coin cause the complete
image to be shown darkly.
The histogram shows that
there are no bright pixels in
the image.
The camera's dynamic
scope will then not be fully
utilized.
2) Move the Exposure adjustment slide control to the right.

" The exposure time for snapshots will then be increased. The dark
areas will then become brighter.
Under the same conditions,
the Exposure adjustment
slide control has here been
moved to the right. This has
lead to a noticeable
improvement in the contrast.
Intelligent Exposure calcu-
lated an exposure time that
was twice as long.
The histogram shows that
the camera's dynamic
scope has been utilized
much better.
9
Calibrating Intelligent Exposure

Before you use it for the first time, you have to calibrate Intelligent Exposure.
The calibration will be automatically set off when you start Intelligent Expo-
sure for the first time.
Intelligent Exposure arranges linear correlation between the exposure time
and the mean intensity for calculating the optimal exposure time. In order to
take advantage of this correlation, the camera's dark current must be known
as an offset of the linear function. Thus, the first step to working with Intelli-
gent Exposure is measuring this dark current via the acquisition of an image
using a darkened camera. When first starting Intelligent Exposure, you will
be automatically asked to carry out this calibration.
ColorView I • Exposure Time Lock
This calibration step measures the camera's dark current, in order to make
an offset correction possible. After successful measurement of the dark
current, Intelligent Exposure asks you to take the next step.
The second calibration step consists of measuring the signals at various

resolution settings. In this way the camera's sensitivity will be determined. A
changing live-image is displayed in this step.
Repeating the The calibration is specific to each camera.
calibration Should you change the camera, you will have to perform the calibration once
more. You have access to this function by using the Calibrate... button
located in the Intelligent Exposure > Options dialog box.
Exposure Time Lock

Exposure Time Click the Exposure Lock button to lock the current exposure time for Intelli-
Lock gent Exposure. From this moment on, no more changes will be made to the
exposure time, even if you shift the Exposure time slide control. Click the
10
button anew to return to the dynamic adjustment of the exposure time.

Use the Exposure Lock option when you want to compare the intensity of
different images or when you want to combine them. You will want to
combine separate images, for example, when you use the MIA add-in, with
which the images are combined into one composite image.
Acquiring images with manual controls

The methods that have been described for making acquisitions using Intel-
ligent Exposure already cover a wide range of possible applications. In
special cases, or when you prefer to have full control over your camera, you
can apply manual controls.
The manual control of your camera offers you numerous possibilities. The
instructions that follow demonstrate a typical working plan. By means of
using different settings for the acquisition parameters you will be shown how
the possible options affect the results you achieve.
Preparing the input channel

1) Use the Image > Configure Input... command.
" The Configure Input dialog box is opened.
2) On the Display tab select the Online histogram check box.
" The online histogram is an important tool that helps you adjust the
camera while the acquisition is being made.
3) In the Image scaling picklist, select the way you want the camera image
to be displayed on your monitor.
" For examining the sample, select the Adjust to viewport option.
With this setting you will see the camera's complete image on your
monitor while you are making the acquisition. It is suitable for
moving to the area of the sample that interests you.
" To focus, select the Full size (100%) option. With this option each
monitor pixel is equivalent to one camera pixel.
4) Click the OK button to close the Configure Input dialog box.
11
ColorView I • Acquiring images with manual controls
Acquiring a live-image
5) Click the Acquire button in the Standard button bar to start the acquisi-
tion.
" The live-image is displayed in the image document.
" The online histogram will be displayed.
6) Click the Camera Control button in the Standard button bar.
" The Camera Control dialog box offers you all of the possibilities for
making settings for the camera.
12
At this point, the exposure time has not yet been correctly set. The histogram shows that the camera's dynamic range
has not been fully utilized. The image appears to be very dark.
7) Click the Auto button, located in the Camera Control dialog box, to have
the exposure time automatically set correctly.
" Alternatively, you can change the exposure time by entering the
values you want, or by clicking the [+] and [-] keys. While doing this,
keep an eye on what the histogram shows.
8) Click the Snapshot button to acquire a snapshot with the current

settings.
The automatic calculation of the exposure time results in the image becoming considerably brighter. The camera's
dynamic range will then be more fully utilized.
13
Automatic Gain Display and fixed scaling

Even when the exposure time has been correctly set, it is still, in certain
cases, possible to improve the image quality. Which of the two methods,
automatic gain display and fixed scaling will achieve the best results,
depends on the sample. Try out both of the methods as described in the
following. The properties of both methods will be explained in the section
Difference between Automatic Gain Display and fixed scaling, p. 16.
1) Click in the Camera Control or Histogram dialog box, the Use automat-
ic gain display button, to activate the Automatic Gain Display. The Au-
tomatic Gain Display regulates the amplification of the signal
continually, so that a fixed proportion will land in the overflow (Channel
0 resp. Channel 4095).
" You can set the proportion of the signal that is to land in the over-
flow by using the Image > Configure Input > Display command.
14
When you activate the Automatic Gain Display, the range that is displayed will be limited correspondingly to the signals
present. In the example shown, it reaches from channel 302 (blue line in the histogram) to channel 3505 (red line).
2) Click the Histogram calculation on ROI to limit the image segment on
which the display is to be optimized.
3) In the Camera Control dialog box, click the Set ROI for Histogram
button.
" A red rectangle will appear in the image. The rectangle indicates
the area that is to be taken into account for the histogram calcula-
tion. To change its size, move the mouse while keeping its left
button depressed, and to change its position move the mouse
without pressing its button.
4) Position the ROI in such a way as to encompass the area that is to be

displayed as well as possible.
Limiting the calculation of the histogram data to that of the red rectangle makes the image appear brighter. The calcu-
lation of the Automatic Gain Display is only based on the data from this rectangle.
15
1) In either the Camera Control or the Histogram dialog box, click the Use
fixed scaling button.
" The Automatic Gain Display will be deactivated.
2) Click the Automatically adjust fixed scaling button to have the once
more specified area adapted to the current conditions.
By having the fixed scaling automatically set, the range displayed will be limited according to the signals present. In
this process, in color mode, only the upper limit (red line) will be changed.
16
Difference between Automatic Gain Display and fixed scaling

Automatic gain display and fixed scaling are both methods of transforming
the crude camera data into an image that can be displayed. While the
camera works with a resolution of 12 bit per color channel, for the display,
storage and further processing, only 8 bit per channel can be used.
Automatic Gain The Automatic Gain Display carries out the adaptation dynamically
Display according to an analysis of the histogram. In this way the image quality will
be automatically adapted when changes are made in the illumination or the
sample. It is, however, not possible to compare separate images that have
been acquired with this setting, where their intensity is concerned.
What counts for the Automatic Gain Display, is the area on which the histo-
gram has been calculated.
Further options in the Camera Control
Button Area Effect

Full image The histogram is calculated on the complete image.
This option is recommended if the structures to be observed
are equally dispersed throughout the entire image.
Crosshairs The histogram is calculated according to a strip that runs in
vertical and horizontal directions in the form of a cross, and is
only a few pixels wide.
ROI The histogram is calculated on a rectangular area that is de-
fined by clicking on the Set ROI for Histogram button.
This option is suitable when the reproduction of a part of the
image is to be optimized.
Fixed scaling With the fixed scaling option the setting that has been made will not be
altered. This makes it possible to compare the intensity of images that have
been acquired with the Fixed scaling option.
This makes it possible to define a fixed intensity range. This method is
mostly used for monochrome acquisitions.

White Balance
The human eye reacts very sensitively to color variations which are
perceived as a color tingeing effect. The adjustment of the color channels
and the white balance depend on the illumination conditions. When
changing the lamp or altering the lamp voltage, the white balance has to be
repeated afterwards.
When using white balance, the individual color channels are scaled in a way
that the white or neutral gray area of the image displayed on the monitor is
displayed correctly as white or gray. The white balance can be carried out
automatically on each image or manually for individual images.
Microscope Settings The microscope settings you use for the acquisition for a white balance must
correspond to those you use in practice. This especially holds true for the
17
filter and the lamp voltage settings. The lamp voltage of a 12V halogen lamp
should be set to 9V. It should, however, not be set lower than 5V. Use the
color neutral ND-filter, should you have to reduce light intensity.
Specialized microscopy It is very difficult to execute a white balance in some microscopic methods,
techniques i.e. DIC or polarization. In these cases, execute the white balance using the
bright field method and only then begin with the special methods.
Transmitted Light When using the transmitted light method, look for a position through which
the light passes through the cover slip, embedding material, and microscope
slide, but not through the specimen. Should such a position not be present,
remove the sample and perform the white balance without a sample. Thus,
the lamp light used for illuminating the specimen falls directly on the objec-
tive.
Reflected light When using the reflected light method, you need a reference surface. Stan-
dardized gray cards are best for this. Should one not be available, you can
also use white laboratory porcelain or neutral ND-filter paper. Should your
samples contain very reflective areas, defocus your microscope when
setting the white balance.
ColorView I • Further options in the Camera Control
Performing a white balance with manual acquisition

1) Acquire an appropriate snapshot.
2) Use the Image > Camera Control... command.
" The Camera Control dialog box is opened.
3) Click the Set ROI for White Balance button.
4) Pull the ROI's red frame shown in the image document to a position
which is a uniform white or gray.
" Change the size of the area by keeping the mouse depressed and
moving the mouse. The ROI used for the white balance cannot be
made to any large size possible.
" The ROI can be difficult to see if the image has a reddish shade. In
this case, use the Special > Preferences > View command to
change the color of the frame. You can select a more appropriate
color from the Primary list located in the Mouse cursor group.
5) Rightclick to confirm position and size of the ROI.
6) Click the White Balance button to conduct an automatic white balance.
" The settings for the individual color channels are optimized in the
set area. The result is directly visible in the image buffer's image.
7) Click the Color Settings button.
" The Color Settings dialog box is opened.
8) Click the Define...button.
" The Define Default Color Settings dialog box is opened.
9) Click the Current settings button to adopt the current values for the rest
of the work.
" When using this setting, the white balance is valid for all images
which are acquired with this input channel during the current
session.
10) Click the OK button to return to the Color Settings dialog box.
11) Click the Standard button to save these values as general standards.
" The current values for the white balance are now saved and will be
imported when the program is restarted. A manual change of the
color settings changes the white balance.
The image analysis program alters the color settings in such a way that the
18
image area in the ROI appears as white or gray as possible. After you have
carried out the white balance once, you can activate it any time to a running
live-image acquisition.
Sharpen filter
Use the sharpen filter to increase the sharpness using edge enhancement
already with the live acquisition.
Sharpen Filter Click the Sharpen Filter On/Off button to activate/deactivate the sharpen
On/Off filter. The manner in which the acquired image is affected by the sharpen
filter depends on the sharpen filter parameter. You can adjust this parameter
by using the Sharpen Filter Settings dialog box.
Clicking the Sharpen Filter On/Off button produces the same results as
selecting/clearing the Activate check box in the Sharpen filter group in the
Input tab. The button is simply a more convenient alternative.
Further information on sharpen filters and sharpen filter parameters can be
found in the section Input tab, p. 40
Sharpen Filter Click on the Sharpen Filter Settings button to open a dialog box where you
Settings can set the sharpen filter parameter:
To adjust the numerical value of the sharpen filter parameter, use the mouse
to move the slide control to the right or left. This is a convenient alternative
to using the Input tab - where you can also adjust this numerical value (in
the Sharpen field).
Color Settings
Click the Color Settings button to manually alter the camera's color settings.
The settings can be altered using the five slide controls: Red, Green, Blue,
Gamma, and Saturation.
Red, Green, Blue The Red, Green and Blue slide controls can each be set from 0,00 to 20,0.
The set value is the factor with which the signal of the corresponding color
19
channel is multiplied.
Gamma Use the Gamma slide control to obtain a nonlinear contrast enhancement.
Please note that this gamma filter does not only affect monitor display but
also alters actual image information. If you are planning on quantitatively
evaluating your images at a later time, use the value 1.0 for the gamma filter.
The Oper menu provides you with commands for adjusting image contrast
at any later time.
What is gamma? As is the case with the slide controls of the individual color channels, the
numerical value for gamma effects how the illumination intensity of the
pixels of the camera CCD chip is converted into the color value for the corre-
sponding pixel in the image buffer. If gamma equals 1, the conversion from
illuminance into gray/color value is linear. If gamma does not equal 1, this
results in a nonlinear contrast enhancement. If gamma is smaller than 1, the
brighter gray values will be spread more significantly than the darker ones.
The result will be enhanced contrast in the lighter areas of the image. The
image as a whole will become darker. If gamma is greater than 1, the darker
gray values will be spread more significantly than the brighter ones. The
result will be enhanced contrast in the dark image areas. The image as a
whole will become lighter.
Saturation The Saturation slide control enables you to alter the color rendering from a
black and white signal to maximum color saturation. In most cases, the
"good" setting of this value is seldom higher than the range of 0,10 to 0,20.
The Gamma slide control's value range is from 0,00 to 2,00; the Saturation
slide control's range is from 0,00 to 0,99.
Default Click the Default button to use the default values defined in the Define
Default Color Settings dialog box from now on.
Define… Click the Define... button to set the standard color settings. The Define
Default Color Settings dialog box is opened.
Factory settings Click the Factory settings button to return the color settings to their original
levels. The factory settings have been adapted in such a way that the color
rendering with a specific equipment configuration is ideal for a large scope
without any additional changes having to be made.
Current Settings Click the Current settings button to make the current values be the future
standard.
Sharpness Monitor
20
The Sharpness Monitor allows you to control the focus settings during a live
acquisition. You are also able to define a rectangular area of the image
which is to be analyzed for the sharpness monitor in the same way as for the
white balance and black balance. The maximum contrast serves as a
monitor for the sharpness in this area. Therefore this area should be limited
to an image segment in which the contrast is a good monitor for the sharp-
ness.
The Sharpness Monitor consists of a dialog box in which a relative measure-
ment of the sharpness is displayed by a changing bar which can be varied
between Blurred and Focused.
If you start a live acquisition with an activated Sharpness Monitor or activate

it during a live acquisition, it will show middle sharpness (50%) in each case.
This is because there is no absolute measurement for the sharpness in an
image and because the sharpness monitor only gives relative values which
are related to the initial situation which was present when starting the live
acquisition.
Focussing and Sharp- The length of the bar changes if you alter the focus settings during acquisi-
ness Monitor tion with an active sharpness monitor. In doing so, the blue marking bar
shows the maximum sharpness reached since the live acquisition was
started. The bar shifts accordingly to the right should the sharpness increase
due to your changes. If the focus settings are already optimal when starting
acquisition, the bar can only shift in direction Blurred. The smallest sharp-
ness value will be indicated by a green marking bar.
Reset to 90% Click the Reset to 90%button to set the sharpness monitor's relative value
to 90%. Utilize this function if you start working with a very blurry image and
if the sharpness monitor's bar reaches the limit to the right while focusing.
By resetting to 90%, the display gets more room for additional focussing.
The Reset to 90% function is also available if the sharpness drops very dras-
tically due to changes made to the microscope settings (i.e. changing objec-
tives) during an acquisition in progress.
Using the sharpness monitor

1) Set all other parameters (microscope settings, exposure time, white
balance) optimally. The application of the sharpness monitor should be
the last step before image acquisition.
2) Acquire a snapshot.
3) Use the Camera Control command.
4) Click the Set ROI for sharpness monitor button to interactively define a
rectangular area of the image for the sharpness monitor.
" To do this, select an area of the specimen for which a good sharp-
ness setting is especially important.
5) Begin a live acquisition.
" The Set ROI for sharpness monitor button becomes inactive.
6) Click the Sharpness Monitor On/Off button to open the Sharpness
Monitor dialog box.
" The sharpness monitor begins with a value of 50%.
21
7) Vary the focus settings and observe the sharpness monitor while doing
so.
" The sharpness monitor bar shows the current sharpness, the blue
markation shows the highest achieved sharpness.
" Mind the exposure time when setting the focus. The sharpness
monitor alters itself with a certain amount of delay when using
larger exposure times (> 100ms). Vary the focus settings in small
intervals and wait each time until the sharpness monitor has been
adapted as well.
8) Find the focus settings by which the sharpness monitor is at its
maximum and use these settings for the acquisition.
Partial Readout
The "Partial Readout" mode is a method of making the live-mode quicker.
For the "Partial Readout" mode, a rectangular part of the image is defined,
to which the readout is limited during the live acquisition. In this way a higher
frame rate is attained, since a smaller amount of data has to be transferred.
The live overlay must be enabled for the "Partial readout" mode. This takes
place on the Display tab in the Configure Input dialog box.
Partial Readout and "Partial Readout" mode and clipping function both lead to smaller images
Clipping than those possible with the camera. The "Partial Readout" mode already
goes into action while the image is being read out from the camera. The
parts of the image that are removed by this function will not even be
exported to your computer. In contrast, a clipping will be not be calculated
until all of the image data has reached your computer. This means that clip-
ping does not accelerate the image acquisition.
Owing to the camera's very high pixel count, when full resolution is being
used, the frame rate is often not high enough to enable good focussing
during live-mode.
Clipping and the "Partial Readout" mode are mutually exclusive. The "Partial
readout" mode is disabled if you enable a clipping on the Format tab located
in the Configure Input dialog box. The ROI which was set for the "Partial
readout" modus remains intact. Clipping is disabled if you enable the "Partial
readout" mode while clipping is enabled. The settings for the clipping will be
preserved. they will be reemployed as soon as you deactivate the "Partial
Readout" mode.
Using the Partial Readout

1) Start the live acquisition using full resolution.
2) Set the ROI for Partial Readout onto an important image detail.
" Change the size of the area by keeping the mouse button
depressed and moving the pointer.
" Change the position of the area by moving the mouse with the
button released.
22
" Fix the area with a right click

3) Activate the Partial Readout.
4) Do the required settings.
5) Deactivate the Partial Readout.
" The image area set with the ROI for Partial Readout applies to the
following snapshot, should the Partial Readout mode be enabled.
6) Acquire a snapshot.
Black balance
The black balance is used for background correction. In an area (ROI) which
you have defined, your image analysis program calculates the mean value
for each color channel. These values are subtracted from all of the pixels in
the entire image. The black balance is to be used predominately for fluores-
cence acquisitions, where it is applied to remove undesired background
from the image. The image changes its color if the black balance is used with
brightfield acquisitions.
Related Topics
Display tab 43
Online shading correction
Using the black balance

1) Acquire a snapshot containing an area that is suitable to be used for a
black balance.
3) Click the Set ROI for Black Balance button to set an area in the image
in which the black balance is to be defined.
4) Drag the red frame that defines the ROI in the image document, to a
position containing an area which is as uniformly dark as possible.
" Change the size of the area by moving the mouse while keeping its
left button depressed. The size you can make this ROI for the black
balance is limited.
" The values for the black balance will then be calculated on the area
you have thus defined.
6) Click the Black Balance On/Off button to activate the black balance.
" The correction values calculated for the black balance will then be
deducted from all of the pixels in the entire image.
The black balance function is available for live acquisitions as well as for
snapshots. A ROI for black balance has to be set as described above before
you perform the black balance on snapshots.
The Black Balance button for snapshots is located in the Black button bar.
Should the Black button bar not be on display, use the Special > Add-In
Manager command to activate it.
Online shading correction

Every optical system with a camera and microscope creates, even when the
devices have been set up very carefully, an effect called shading.
The left illustration shows an
image where the online
shading correction has not
been activated. The image
intensity decreases towards
23
the edges.
Shading Correction For a shading correction, this effect will first be determined to enable correc-
tions to be made to the acquired images. Using the online shading correc-
tion of ColorView I, the shading correction already takes place in the live-
image.
ColorView I • Using the Shading Correction
Using the Shading Correction

When your microscope had been precisely set up, you will hardly be able to
perceive any shading where snapshots are concerned. The use of a shading
correction may, in this case, be advisable, but is not essential. When,
however, several snapshots are combined, as for example, when you use
the MIA add-in, then even mild shading will immediately become visible.
How the Shading Correction functions

The correction of the image takes place pixel bx pixel with the help of two
different reference image series: one made up of dark images for each of
the camera's resolutions, the other of correction images. The online shading
correction will not be ready for use until you have acquired these reference
images.
Dark Images A dark image is an acquisition in which no light falls on the camera. Noise or
defective pixels may here produce signals which can be corrected by the
dark image. Each of the camera's resolutions requires its own dark image.
Correction Images The illumination of the entire optical system without the object (with refer-
ence samples in reflected light mode) is shown. In addition to the camera
characteristics, the microscope's optical characteristics, especially the
objective being used, contribute to the correction image. Accordingly, a
correction image must be acquired for each camera resolution and objective
combination.
Acquisition Methods As well as the already mentioned influences that camera and objective
exert, the acquisition process you use also plays a vital role. The online
shading correction will have been aligned with transmitted light, reflected
light, DIC and Phase Contrast microscopy. When acquiring the correction
images, you will be asked which acquisition processes have been set.
Should you want to employ more than one process, you will have to repeat
the acquisition of the correction image for each process used. If, while you
are using the program, you change the acquisition process, you will also
have to change the Online Shading-Correction's setting.
Note When using a magnification changer, correction images must be acquired
for each possible total magnification.
24
Acquiring Reference Images

Note Before you acquire reference images and activate the shading correction,
make sure that your microscope is correctly adjusted and that the objective
is clean.
Please be very careful when acquiring reference images for the online
shading correction, since false correction acquisitions can lead to artifacts in
the images. The actual acquisition process is controlled by a wizard. This
wizard tells you which steps to take.
Prerequisites for a refer- A reference acquisition always relates to one particular magnification, for
ence acquisition this reason, before the reference image is acquired, an XY-calibration of the
objective to be used will have to be performed. The calibration that you can
perform when your image analysis program is installed, fulfills this require-
ment.
Related Topics
Illumination method 41
Acquiring Reference Images
As part of the preparations for making the acquisition, the wizard asks you
if the correction acquisition is to be made only for the current magnification
or for all calibrated magnifications.
Transmitted Light 1) When using a tungsten-halogen lamp, set the same lamp voltage as
Mode when using normal operation.
" The lamp voltage should not be less than 5V. Use ND-Filters to
reduce possible brightness.
Note ND-Filters belong to the basic components of the microscope. They will be
brought into the light path to reduce brightness without altering the color
temperature. Should your microscope not have an ND-filter or not provide
for the installation of one, you will instead have to reduce the lamp voltage.
2) Set the illumination method to brightfield. This also holds true if you
want to use special methods at a later time.
3) Bring the objective with the smallest magnification, by which the
condensor front lens still has to be used, into the light path and set the
Köhler illumination.
4) Set the camera's exposure time to a value of > 50ms by lowest resolu-
tion.
" The value in the online histogram should be around 2,000.
The online histogram shows
the correct setting for the
acquisition of the gain
image.
5) Bring the object into the light path and bring it into focus.
6) Defocus by increasing the space between the objective and the object
until the object is recognizable as a whole but the details of the struc-
ture are no longer visible.
25
7) Move to a position on the slide where neither the sample is visible, nor
are there any scratches or contamination.
" Should such a position not be able to be found, remove the micro-
scope slide from the microscope and acquire the reference images
without the object.
8) Begin the acquisition of the reference images with the Image > Acquire
Reference Images... command.
" The Online Shading-correction dialog box will open.
9) Follow the wizard's instructions to acquire the reference images.
Reflected Light Mode 1) When using a tungsten-halogen lamp, set the same lamp voltage as
when using normal operation.
" The lamp voltage should not be less than 5V. Use ND-Filters to
reduce possible brightness.
Related Topics
Köhler Illumination 50
ColorView I • Acquiring Reference Images
Note ND-Filters belong to the basic components of the microscope. They will be
brought into the light path to reduce brightness without altering the color
temperature. Should your microscope not have an ND-filter, please contact
your supplier.
2) Set the illumination method to brightfield. This also holds true if you
additionally want to use special methods at a later time.
3) Bring the objective with the smallest magnification, by which the
condensor front lens still has to be used, into the light path and set the
Köhler illumination.
4) Set the camera's exposure time to a value of > 50ms by lowest resolu-
tion.
" Your online histogram's gray value should be around 2,000.
The online histogram shows
the correct setting for the
acquisition of the gain
image.
5) Bring the reference sample into the light path and set the Köhler illumi-
nation.
" The reference sample must be color neutral and have a flat surface
devoid of defects or optical structures visible through the micro-
scope.
Note Should you not have access to a reference sample, you can also use a piece
of opaque glass. In this case, take special care that you move it out of focus.
6) Increase the distance between the objective and the reference sample
until the surface structures of the reference sample are no longer
visible.
Warning Please note that the focus depth is high when you use objectives with lower
26
magnification. In this case, you have to defocus more than with objectives
with a magnification of 10x or higher.
When using reference samples with transparent surfaces, it is important to
be sure that no plane within the reference sample is in focus.
7) Begin the acquisition of the reference images with the Image > Acquire
Reference Images... command.
" The Online Shading-correction dialog box will open.
8) Follow the wizard's instructions to acquire the reference images.
9) Carry out a new white balance when you first use the online shading
correction.
Related Topics
Köhler Illumination 50
Enabling Shading Correction
Enabling Shading Correction

When you have finished acquiring the reference acquisitions, you will be
asked if you want to activate the online shading correction. Independent of
the decision you make right then, you can activate or deactivate the online
shading correction on the Image > Configure Input > Input tab at any time.
The settings of the online shading correction remain even after the program
has been restarted.
Input channel and The shading correction's reference images are valid for one particular
shading correction camera. They will also be used for other input channels that have been set
up for this camera. The fact, whether the online shading correction has been
activated or not, is, on the other hand, a property of the input channel. Where
newly set up input channels are concerned, the shading correction is not
activated by default. In the case of a duplicated input channel, the shading
control's setting will be adopted from the input channel that has been dupli-
cated.
Shading correction when working in black & white

The shading correction is also available to you when you are working in
black & white. Should reference images for the online shading correction
have been acquired while you were working in color, these dark images will
also be valid when you work in black & white. The correction images
however, have to be acquired again.
27
Related Topics
New Channel 39
ColorView I • Acquiring Reference Images
Repeat or Update the Reference Acquisitions

The online shading correction functions optimally as long as the illumination
settings are not changed.
Sample acquisitions Sample acquisitions enable you to check the quality of the online shading
correction. To do this, acquire an image of a sample with a background that
is as even as possible. Then measure the acquired sample's intensity distri-
bution with the commands in the Measure >Intensity Profile submenu. When
the shading correction has been correctly set, the intensity profile will,
except for the sample's structures, be even.
Example of an intensity
profile without shading
correction (or with a poorly
performed one), and with
shading correction.
The intensity profile shows a
clear decline along the X-
axis.
28
Alterations to the illumi- If you switch to another illumination method which is supported by the online
nation method shading correction, you may have to make new correction acquisitions.
Should the required acquisitions already exist, you can set the appropriate
illumination method with the Image > Configure Input > Input command. To
do this, select the appropriate entry from the Illumination method list.
Adding/changing objec- If you add new objectives or change already installed objectives, you must
tives acquire correction images for the new objective.
Altering the lamp voltage Small alterations made to the lamp voltage do not influence the online
shading correction. Larger alterations cannot be compensated by the online
shading correction. Therefore, use the same lamp voltage you utilize in your
normal routine use, when you acquire the correction images. Should you
need to reduce the brightness, use ND filters. Otherwise you will have to
acquire new correction images for the shading correction.
Use of the Online Shading Correction
Use of the Online Shading Correction

Please note the following instructions for the use of the online shading
correction in order to be able to make optimal acquisitions.
Before acquisition Make sure that the optical system is clean. Check the lamp voltage and
check if the Köhler illumination has been set. Furthermore, there should not
have been any changes made on the microscope's light path since the last
reference images were acquired.
Setting magnification The correct magnification must be set for the use of the online shading
correction.
Automatic microscope It is not necessary to set the magnification when using an automatic micro-
scope which is remote controlled by the application.
Technical Note
Organizing the reference Dark images and correction images are saved to the hard disk. In order to
images be able to work with a correct online shading correction when having
changed the camera, the image analysis program creates an individual
directory for saving the reference images. This directory is in the image anal-
ysis program's working directory. Its name begins with DGSISFW and
continues to contain information about the serial number of the camera
being used. This directory should not be moved or renamed.
The reference images file names are also coded in such a way that the
online shading is able to automatically access it. Should one or more of the
reference images have been deleted, the application immediately tells you
that a missing image is required and automatically requires you to reacquire
the missing reference image.
File Format The reference images are saved as normal TIF images.
Requirements The filter in front of the camera's CCD chip and all of the microscope's
optical parts must be clean. Otherwise, dust particles or fingerprints would
appear in the correction images. The shading correction can, of course,
correct such contamination, as long as they remain unchanged. Should a
contaminant be moved or removed, the shading correction would give you
a ghostly image of the former contamination in each new image. Check the
29
filter before acquiring the reference images and clean it with a LensPen, if
necessary.
intX and Online Shading Correction

The Intelligent Exposure acquisition method works problem-free together
with the online shading correction. In some cases, however, the shading
correction can result in over exposure. You will then receive an error
message when executing a white balance.
Adapting Intelligent In this case, click the Options...button located in the Intelligent Exposure
Exposure dialog box. Reduce the Dynamic limit in %value in the Intelligent Exposure
Options dialog box to approximately 70.
ColorView I • Using the black & white mode
Using the black & white mode

In this section, the control elements in the Camera Control dialog box that
are different from those in color mode, will be described.
For the black & white mode a separate input channel is needed. When the
camera and software are installed this channel will be automatically set up.
Enabling Black & White Use the Image > Set Input... command to select the input channel which was
mode created for the black & white mode.
Gamma Correction
In areas with rather high and rather low intensities, cameras can come to
non-linearities which can decrease the contrast of these areas. You can use
a gamma correction to make a physiological correction.
Gamma Setting Click the Gamma preferences button to set up a quick gamma correction
during an acquisition. The Gamma Setting dialog box is opened.
30
Move the Gamma slide control to make a gamma correction of the live-
image. Move the slide control to the left (Gamma < 1), to make the entire
image darker; At the same time, the contrast in bright areas of the image will
be increased. Move the slide control to the right (Gamma > 1) to make the
entire image lighter and to increase the contrast in the dark areas of the
image.
Note The image will be displayed using an LUT (Look-Up Table), or in other words
is converted into a false-color image. For this reason, not all of the analyses
are still possible, e.g., phase analysis.
Black balance
The black balance is used for background correction. The program calcu-
lates the mean value for each color channel in an area (ROI) which you
defined. These values are subtracted from all pixels in the entire image. The
black balance is to be used predominately for fluorescence acquisitions in
order to balance the undesired background in the acquisition.
Acquisitions with Real time FFT
Performing a black balance

1) Take a snapshot which contains a suitable area to be used for black
balance.
3) Click the Set ROI for Black Balance button to set an area in the image
in which the black balance is to be defined.
4) Pull the ROI's red frame shown in the image document to a position
which contains an area which is as uniformly dark as possible.
" Change the size of the area by keeping the mouse depressed and
moving the mouse. The ROI used for the black balance cannot be
made to any large size possible.
" The values are calculated for the black balance on the area defined
in such a way.
6) Click the Black Balance On/Off button to activate the black balance.
" The correction values calculated for the black balance are
deducted from all the pixels in the entire image.
The black balance function is available for live acquisitions as well as for the
acquired snapshots. A ROI for black balance has to be set before
performing the black balance on single images. The Black Balance button
for acquired images is part of the Black button bar.

When working in black & white you can use a Fast Fourier Transformation carried out in real time to check
the image quality in certain cases.
To be able to utilize Real time FFT during a live acquisition, you have to
select the Image > Configure Input > Display > Live overlay check box.
Enabling Real time FFT Real time FFT can be activated in the input channel. To do this, select the
Image > Configure Input > Display > Real time FFT dialog box.
Real-time Fast Fourier Transformation (RTFFT) calculates the acquired
image's Fourier image in real time (the absolute value of the Fourier trans-
31
forms). The Fourier image is continually updated during a live acquisition. It

can serve as a quantitative display of the image quality. For example, if you
blur the microscope image, the high frequencies disappear in the Fourier
image (i.e. the structures which are located farther away from the center).
ColorView I • Acquisitions with Real time FFT
The Fourier Image of a

Catalase 2-D Crystal.
The points represent peri-
odic structures in the micro-
scope image. The closer
such a point is to the center,
the smaller the spatial
frequency of the corre-
sponding periodic structure,
i.e. the larger the periodic
distances in the microscope
image. (Fringe effects cause
the cross to appear.)
To achieve optimum on-screen display of the fourier image, use the RTFFT
Control dialog box with the tabs ROI, Display, and Tools.
ROI
Use the ROI tab to select the area, on which the RTFFT is to be carried out.
The RTFFT is calculated on a square segment of the image. Two different
32
sizes are available for the calculation. You can also position the square that
is to be analyzed anywhere in the image.
Note In case the sizes offered you are still too small, you can activate binning.
This increases the size of the ROI compared to that of the image segment.
256x256, 512x512 Select the option 256x256 or 512x512 to make the ROI the respective size.
Show ROI Select the Show ROI check box to display the position of the ROI in the live-
image. The ROI is displayed as a red frame.
Position Click on the Position button to move the ROI in the live-image. The RTFFT
Control dialog box will be hidden and the pointer appears in the live-image.
The red frame shows the position of the ROI for the RTFFT. Move the
mouse to alter the position of the marked area within the image document.
Click the right mouse button to fix the position of the ROI.
Display
Use the Display tab to modify the monitor display of the RTFFT.
Contrast The Contrast slide control enables you to alter the contrast of the display in
the RTFFT Display window. Move the controller to the right to increase the
contrast and to the left to decrease it. Should you have selected a Display
palette the colors will change in the RTFFT Display window. You define the
palette in the input located on the Display tab.
Brightness The Brightness slide control enables you to alter the brightness of the Real
time FFT display in the RTFFT Display window. Move the controller to the
right to increase brightness. Move the controller to the left to decrease
brightness. Should you have selected a Display palette the colors will
change in the RTFFT Display window.
Reduced FFT Select the reduced FFT check box to further reduce the image section on
which the FFT will be calculated. At the same time the display in the RTFFT
Display will be enlarged.
33
Tools
The Tools tab offers RTFFT-related tools.
90° Rotation Select the 90° Rotation to alternate the FFT display between the usual orien-
tation and one rotated through 90°.
Overlay Circle Select the Overlay Circle check box in order to show a marking circle in the
RTFFT Display window. The circle will be shown in the center of the window.
With its help you can position the camera or sample.
Related Topics
Display palette 47
ColorView I • NetCam
Circle Size Move the Circle Size slide control to the right to increase the diameter of the
overlay circle. Move the Circle Size slide control to the left to decrease the
diameter of the overlay circle. The circle size is represented in pixels in the
RTFFT Display window.
Grab FFT Click the Grab FFT button to save the content of the RTFFT display window
as a single image. The image will be written into the next image buffer. This
image's name is that of the live-image, supplemented by 'FFT(8bit)' and a
consecutive number.
When you acquire a snapshot as a conclusion of the live acquisition, such an
FFT image will also be created and saved.
NetCam
You can export the ColorView I live-image to other computers via an intranet
or the Internet, by using the NetCam add-in.
This enables you to make the images immediately available to your
colleagues during an acquisition. This makes it possible for you to work
interactively over large distances by using simultaneous communication,
e.g., telephone. The images will be compressed in the process to speed up
the rate of transmission.
Transferred images The live-image will be directly transferred. Provided your network connec-
tion achieves an adequate speed and the frame rate attained during acqui-
sition, is sufficient, you will get the impression of a live-image. In an internal
company network, an immediate feedback without any time delay will be
possible between observer and user.
NetCam can also transfer a fixed image, whereby in this case the image's
overlay will also be transferred.
What is a Server? A Server is the computer on which both the image-analysis program and the
NetCam are installed. The image acquisition is controlled from the server.
What is a Client? A Client is any computer that is able to access the server's images via the
Internet. The client cannot control image acquisition.
A web server must be installed on your computer to make images available
via the Internet. An appropriate program will be installed during the setup, if
34
you have selected the NetCam add-in. This step will be skipped should a
web server already exist on your computer.
Warning When you use a computer connected to a network it is possible that the
security settings will not allow its use as a web server. In this case, contact
your system administrator.
In your image analysis program's Module\NetCam subdirectory you will find
the file "NetCam_Info.pdf". This file contains technical explanations about
NetCam which you can give to your system administrator.
Installing the web server
Installing the web server

The web server you require for using NetCam will be automatically installed during the setup.
The installation of the web server is started directly as part of the setup. After
you have agreed to the license terms, you can make further settings in a
dialog box.
You can simply adopt the default settings.

Complete, Custom We recommend the complete installation of the web server.
NetCam in the intranet

35
The transfer of images within the company network or intranet generally

does not require any further actions.
Enabling access During installation, the setup will create a link with the name NetCam.html in
the \Module\NetCam subdirectory below your image-analysis program's
working directory. Send this link via e-mail to colleagues you would like to
give access to your images. The easiest way to do this is with the following
command: Send to > E-mail receiver in the context menu of the link in
Windows Explorer.
Prerequisites for clients Java Runtime Environment, Version 1.4.1_03 or higher is required to show
the images on the clients. When this requirement has not been fulfilled by
the client, he will not be able to receive the images.
ColorView I • NetCam in the internet
NetCam in the internet

When using NetCam, a direct communication takes place between the
server and the client. If the communication goes beyond the company
network, you must, if need be, make changes to the security settings.
Adapting the security The adaptation of a network's security settings takes place via the respec-
settings tive system administrator. The necessary technical and security require-
ments for the use of NetCam are outlined in a separate file entitled
"NetCam_Info.pdf" which will have been saved during setup to your image
analysis program's \Module\NetCam subdirectory. Please give this file to
your system administrator so that he can make the appropriate adaptations.
IP Address The server's internet address must be entered on the client's page. This
address is available in the well-known alphanumerical form ("MyCom-
puter.OurCompany.de") and in the numerical form ("62.180.61.137"). The
numerical form of the address is more reliable, especially if your system
administrator has to alter the network settings.
Determining the IP You can find out your computer's IP address from your system administrator
address or determine it yourself.
Determining the IP address

1) Select the operating system command Start > (All) Programs > Acces-
sories > Command Prompt.
" The Command Prompt console box will be opened.
2) Enter the ipconfig command via your keyboard and use the enter key.
" Data about your computer's network connection are shown in the
console box.
The iconfig command will
display a computer's IP
address
36
3) Note the entry in the IP address row.

4) Supply the users who are to have access to your server as NetCam
clients with this IP address.
Prerequisites for clients Java Runtime Environment, Version 1.4.1_03 or higher is required to show
the images on the clients. When this requirement has not been fulfilled by
the client, he will not be able to receive the images.
Using NetCam
Using NetCam
The use of your image-analysis software remains the same when using
NetCam. You can use all of the functions as always. To transfer the images
to other users and to be able to control the transfer, use the buttons located
in the NetCam button bar.
NetCam on/off Click the NetCam On/Off button to start or end the transfer of images with
NetCam.
The transfer of live-images begins as soon as NetCam is activated. The
transfer continues until you deactivate NetCam or close your image-analysis
software.
NetCam Click the NetCam Settings button to set image options and to administer the
Settings clients which are to have access to your images. The NetCam Settings
dialog box is opened.
Connections
Use the Connections tab to administer the clients which are to access your
images.
37
Connected Clients In the Connected Clients list you will see all of the clients which are currently
connected with your computer. You can mark one of these clients to discon-
nect the connection to it or to refuse a connection in the future.
Disconnect Click the Disconnect button to disconnect the connection to the marked
client. The client can once again make a connection.
>> Click the >> button to include the marked client in the Banned Clients lists.
The corresponding client is automatically disconnected.
Banned Clients In the Banned Clients list you will see all of the clients who you have
forbidden access to your image-analysis software. From these computers it
is impossible to load and view the images made available by NetCam.
Add new... Click the Add new... button to add computers or entire groups of computers
to the Banned Clients list. The Enter IP(s) or a domain dialog box is opened.
ColorView I • Using NetCam
Domain name Enter the alphanumerical name of a domain in the Domain name field if you
want to block the access of all computers belonging to a certain domain. For
example, enter the domain name foobar.com to block the transfer of the
images to all of the computers within the foobar.com domain.
Procedure NetCam sends a query to the nameserver to get the numerical IP addresses
belonging to the specified domain. Should the query lead to an error, you will
receive a message. The specified domain will, in this case, not be placed on
the Banned Clients list.
IP Address(es) You can provide a numerical IP address or an entire address range in the IP
address(s) group to exclude the computers in question.
From, to Enter the limits of the address range to be excluded in the from and to fields.
38
Image options
In the Image Options tab you can set the quality of the transferred images
and thus influence the transfer rate.
Image quality Use the Image Quality slide control to control the compression of the
images. Move the slide control to the right to maintain a high image quality.
The data amount of the transferred images increases, whereby the transfer
rate decreases. Move the slide control to the left to increase the compres-
sion. The image quality decreases, whereby the transfer rate increases.
Image quality and data A single image of ColorView I can be of a size up to 15 Mbyte. With an image
transfer rate of this size, the transfer to a client can take a long time, even if you're using
a very quick network connection; the effect of a live-image being transferred
will be lost. For this reason, images are compressed for transfer. The algo-
Setting up the camera
rithm is equivalent to that used with JPEG images. Thus, high compression
rates are achieved, whereby, however, image quality decreases.
The amount of compression achieved is based on the image type. Images
with large, uniformly colored areas can be compressed more than detailed
images without these characteristics.
Setting up the camera

Logical input channels
For every camera that is to be used with the image analysis program, one
or more logical input channels will be set up. Logical input channels contain,
for example, the information about how large an image format is, and what
is to happen with the image signal before it reaches the image buffer. Each
channel must be calibrated separately. A channel for image acquisition via
ColorView I is usually called "ColorView I FW...".
Status after installation Two input channels are automatically created when the software is installed,
their settings enable you to immediately acquire images. One channel uses
your camera's black & white mode, the other its color mode.
Precalibration During the installation you will be offered the opportunity of performing an
XY-calibration by entering the objectives and the TV adapter to be used.
Thanks to this automatic calibration you can then acquire calibrated images
without any further preparations.
Configuration via tabs Input channels are configured via the inputting of suitable values in various
tabs. The 3 tabs relevant for ColorView I will be dealt with in-depth.
Setting up additional You can set up and use more than 100 additional input channels. This can
input channels be useful when you want to employ different settings for a variety of uses. If
you have to set up a new input channel, you can create a copy of an already
existing input channel and adapt it, or create an entirely new input channel.
Set Input Use the Image > Set Input... command to select a new input channel or to
create a new one. You can also depress the [F6] key. The Set Input dialog
box will be opened.
The Set Input dialog box
39
with an already defined

input channel.
New Channel Click the New Channel button in the Set Input dialog box to create an entirely
new input channel. The Select device dialog box will be opened. All
connected image creating devices are listed in the Available devices list.
ColorView I • Input tab
Select Device dialog box

with a ColorView I camera
connected.
The abbreviation BW in the
last line indicates this
camera's black & white
mode.
Select the desired camera mode (color or black & white) in the Available
devices list and click OK in order to create a new channel.
Creating additional input Should you want to create additional input channels, for example to have
channels different configurations immediately available for repeating tasks, use an
already existing input channel.
Duplicate Click the Duplicate Channel button to make a copy of the active input
Channels channel. The new input channel gets the same name as the channel from
which it was copied and receives a sequential number. The newly created
channel is automatically enabled.
Configure Click the Configure Input button to adapt the settings for the new channel.
Input The Configure Input dialog box will be opened. You can also open this dialog
box by using the Image > Configure Input command or by double clicking
the camera name located in the status bar.
Input tab
40
Input tab
Exposure Enter the number of microseconds the camera's CCD chip is to be exposed
in the Exposure field. You can also use the arrow key (to the right of the field)
to select this value. The chapter entitled "Camera Control" explains how you
can change the value in this field with the help of the Camera Control...
command interactively while viewing a live-image. The shortest possible
exposure time is 100 µs and the longest is 160 s.
Resolution You can choose one of four different resolutions that are possible with the
ColorView I, in the Resolution list.
Online shading You can set the camera function with the same name in the Online Shading-
correction correction group.
Activate Select the Activate check box to use the online shading correction. The
online shading correction will immediately be used should correction images
already exist for the current settings. Should there not be appropriate correc-
tion images available, the application will automatically start the wizard for
the acquisition of correction images.
Illumination method Select the procedure you use in the Illumination method list.
Sharpen filter Select the Activate check box in the Sharpen filter group to enhance the
image sharpness. This takes place in real time during image acquisition.
When applying the sharpen filter, the numerical value in the Sharpen field
determines whether or not the image’s contours appear sharper or softer.
Raising sharpness will accentuate edges, but will also increases image
noise. Reducing the sharpness makes the image softer. If you enter a value
of 0, the sharpen filter will have no effect at all.
Please note that when you use the sharpen filter you not only alter the
monitor display, but the actual image information as well. This means that
when you are planning on quantitatively evaluating your images later, you
should not use the sharpen filter. The Oper menu provides you with
commands for adjusting the sharpness of an image at any later time.
Convenient alternatives for activating/deactivating the sharpen filter, as well
as for adjusting sharpen filter parameters, are the Sharpen Filter On/Off
button and the Sharpen Filter Settings button - both in the Camera Control
dialog box.
Mirror The Mirror group enables you to determine if the camera's image is to be
41
mirrored directly into the image analysis program during transfer. Both
mirror functions can be combined with each other.
Horizontal Select the Horizontal check box to mirror the image horizontally during
acquisition.
Vertical Select the Vertical check box to mirror the image vertically during acquisi-
tion.
Info Click the Info... button to have a variety of device data displayed in the Color-
View I FW Information window.
Related Topics
Acquiring Reference Images 24
Acquiring images with manual controls 11
ColorView I • Input tab
Device Name The Device Name relates to the inner construction of the camera and is
specified by the manufacturer.
Devices ID The Device ID is the physical identification of the camera for the operating
system. It is in accordance with the device's serial number.
Firmware Version The Firmware is the cameras software for your operation.
Serial Number The serial number serves as a clear identification for each camera
produced.
Camera Type The Camera Type is the exact identification for the camera model.
DLL Version The DLL Version is the version number of the camera's driver.
SYS Version The SYS Version is the version number of the camera's device driver. DLL
and SYS Version are identical.
Temperature The cameras are equipped with temperature sensors. The temperatures are
continually measured.
CCD Chip CCD Chip is the CCD chip's temperature. An increase in noise is the result
of high temperatures of the CCD Chip. The default value for trouble-free op-
eration is 10°C.
Housing Housing is the temperature of the cameras housing. The cameras housing
42
should not exceed 40°C.

Display tab
Display tab
Over exposure The Over exposure group enables you to determine if and when you should
be warned of an overexposure. The software continually checks the image
intensity in live mode, by analyzing the image's gray-value histogram in real
time. You will be warned if the live-image is overexposed. The warning will
also appear following acquisition of an overexposed single image.
Display warning Select the Display warning check box to have a warning message shown if
the image is being overexposed:
43
In this case, reduce the exposure time till the warning message disappears.
Overflow An image is overexposed when a certain percentage of its pixels attain the
maximum gray value possible. This percentage is entered into the Overflow
field.
Automatic gain display Use the functions in the Automatic gain display group to have the image
always displayed optimally on-screen, no matter what the actual lighting
conditions are. The system assesses the current gray-value histogram - in
real time - in order to obtain optimal display of the live-image on the monitor.
Please note that the Automatic gain display not only alters the monitor
display, but the actual image information as well. If it is activated, the reduc-
tion of the acquired image from 3 x 12 bit to 3 x 8 bit takes place with the
help of optimized lookup tables. If the Automatic gain display is not acti-
vated, the 4 lower bits (from the 12 bits of each component color) will be
clipped.
ColorView I • Display tab
Activate Select the Activate check box to enable the Automatic gain display. The
effect of this check box is the same as clicking the Use automatic gain
display button located in the Camera Control dialog box (<). The Activate
check box is not available if the check box with the same name in the Fixed
Scaling group is enabled.
Left overflow When the automatic gain display is active, image intensity for the image on
Right overflow the monitor will be spread between the gray values Gmin and Gmax. You
determine the values of these Gmin and Gmax gray values in the Left over-
flow and Right overflow fields: this is where you enter the percentage of
pixels with the lowest gray values which are to be displayed black, and which
percentage of the highest gray values are to be displayed white.
This is how you clip gray values that are to be ignored at both the upper and
lower ends of the gray-value histogram. Make use of the overflow values to
stop individual pixels or minute image areas that are too bright or too dark
from influencing the Automatic gain display.
Note The left limit can only be set in black & white mode; using it with color
cameras would lead to a color tinge.
Fixed scaling Instead of using the Automatic gain display group, you can use the Fixed
scaling group. The group whose Activate check box has been selected will
be the one available - the other will not be available.
The functions of the Fixed scaling group are for manually influencing on-
screen image display. You select definite limits - Gmin and Gmax - for the
gray-value range that you wish to have optimally spread for on-screen
display. Any gray values occurring outside this defined gray-value range will
appear either black or white.
Application This is how you can, for example, improve the visibility of the specimen
structures of interest to you - by having dark areas of the specimen
displayed entirely in black.
Please note that this manual presetting of the gray value, like the Automatic
gain display, not only effects the monitor display, but also the image infor-
mation.
Activate Select the Activate check box to switch on the manual definition of the gray-
value range to be displayed.
44
Left limit The left limit can only be set in black & white mode; using it with color
cameras would lead to a color tinge. Enter the value of the left limit for the
fixed scaling into the Left limit box.
Right limit Enter the value of the right limit for the fixed scaling in the Right limit box. Its
function is equivalent to the Right limit parameter, which you can influence
by clicking the Manually adjust fixed scaling button located in the Camera
Control dialog box.
Online histogram Select the Online histogram check box to enable you to keep an eye on the
gray-value distribution during image acquisition. During image acquisition, a
window showing the current histogram will appear automatically. This histo-
gram will be continually updated.
Display tab
Online histogram while

operating in color mode
(above) and in black & white
mode (below).
Minimum, mean, and maximum gray values of the image will be shown
beneath the histogram. Percentages are in relation to the maximum obtain-
able gray value. The maximum obtainable gray value when using ColorView
I (with its 12-bit depth) is 4095. This gray value is equivalent to 100%. A
mean value of 19,2% when operating in black & white is equivalent to a gray
value of 788.
During a live acquisition, you can turn the Automatic Gain Display on or off
with the buttons in the online histogram, as well as activate and set the fixed
scaling.
Setting the fixed scaling You use the mouse to set the limits of the fixed scaling in the online histo-
via the online histogram gram. Move the pointer over the blue (lower limit) or red (upper limit) vertical
line in the histogram. The pointer turns into a horizontal double arrowhead.
Move the mouse with the mouse button depressed to the left or right to either
increase or decrease the limit values. The changes can be viewed directly
45
in the live-image.
Real time FFT The Real time FFT check box is only available in black & white mode. In
order to be able to use the Real time FFT during live acquisition, you must
select the Live Overlay check box. An explanation of the effects of the
enabled Real time FFT can be found in the section entitled Display and
usage of the acquisition with Real time FFT.
Live overlay Select the Live overlay check box if you want to use overlays in live-images.
Live overlays are absolutely necessary for "Partial readout" mode, white
balance, or Real time FFT.
An image’s overlay contains information in the form of text, markers, or other
graphical elements. The overlay can be displayed together with the corre-
sponding image without the actual image information being affected. The
live overlay must be enabled in order to make Real time FFT or "Partial
readout" mode possible.
Related Topics
Automatic Gain Display and fixed scaling 14
Partial Readout 22
ColorView I • Display tab
The Live overlay check box is only available if your display board features a
so-called hardware overlay which is supported by Microsoft DirectX. Almost
any up-to-date AGP display board provides you with that feature. You can
look at the System Info dialog box to check whether your display board
offers this feature. To do so, select the Info... command from the ? menu and
click on the System Info... button. The System Info dialog box will be
opened. Click on the "DirectX" entry in the list on the left-hand side to have
the desired information appear in the list on the right-hand side.
Displaying the histo- Select the Show histogram ROI check box to have the ROI for the histogram
gram ROI displayed during the live mode.
Image scaling Select one of the four selections for displaying an acquired image in a View-
port in the Image scaling list.
Underscan Select the Underscan entry to have the whole image displayed. The system
will select the largest zoom level - 25%, 50% or 100% - where the whole
image can still be displayed within the Viewport. In certain cases not all of
the Viewport’s available area will be used.
Overscan Select the Overscan entry to ensure that all the Viewport’s available area will
be used. The system will select the smallest zoom level - 25%, 50% or 100%
- where the image fills the whole Viewport. In certain cases parts of the
image will not be visible.
Adjust to viewport Select the Adjust to viewport entry to have the image size adjusted to opti-
mally fit the Viewport. The image will be adjusted to fit the current size of the
Viewport.
Full size (100%) Select the Full size (100%) entry to have the image displayed without any
zooming. Only the left upper corner of the image will be shown if the View-
port is smaller than the image.
Histogram calculation Select the pixels to be taken into consideration for calculation of the gray-
value histogram in the Histogram calculation list. There are three options:
Full image If you select the Full image entry, the gray values of all pixels will be applied
to histogram calculation. This is the most commonly used setting when
using light microscopy.
Crosshairs If you select the Crosshair entry, only pixels of a particular image area are
used in the calculation. This image area is shaped like cross hairs that are
46
centrally positioned on the image. The vertical and horizontal segments of

these cross hairs are respectively just a few pixels wide and cover just about
the entire height and width of this image. Crosshair is the default setting
when using transmission electron microscopy.
ROI Select the ROI entry to carry out the optimization of contrast and exposure
time based on a rectangular frame.
These settings affect all of the real time functions that use the gray-value
histogram: the warning for Over exposure, the Automatic gain display and
the Online histogram.
Application We recommend selecting Crosshair if you wish to speed up histogram
calculation. You can speed up the frame rate of the live-image depending on
what real time functions you have activated. You should however make sure
that the pixel gray values around the middle image line and around the
middle image column are representative for the gray values of the entire
image. If this is the case, you will not notice any difference compared to the
Full image option for the over exposure warning, Automatic gain display,
and the online histogram.
Format tab
Display palette You can select a false color lookup table from the Display palette list to color
the acquired image. The Display palette list is only available in black & white
mode.
- none - Select the entry - none -, if you don't want to use any palette at all for the
image display.
Adjust, Dither, Gamma, The entries in this list are the names of various lookup tables which can be
Mixmap Rainbow, used for the image display on the monitor.
Solaris, Thermal
Format tab
Active area The display in the Active area field shows how large the acquired image
segment is compared to the largest possible image, when the current
47
settings are applied. If you make changes to the clipping, you can directly
view their effect here on the active area.
Image format You can choose a means of image acquisition from the Image format list.
The entries in this list correspond to predetermined settings for the clipping.
In addition to the predetermined list elements, you can also select from the
settings you have defined.
Image resolution and With the ColorView I you can influence the size of the images in pixels in two
Clipping ways. In the Image format list the sizes of images that can be achieved by
cutting off the edges (Clipping) are contained. In the Resolution list located
on the Input tab you can select resolutions that achieve the same image
format in pixels with a constant display area.
Binning The Binning function is not enabled when you use the ColorView I. Use the
Resolution function located on the Input tab instead.
Clipping border (pixel) When clipping is used, the edges of the images are cut off. Hence, clipping
does not have any effects on the resolution and brightness nor on the
required exposure time.
Related Topics
Resolution 41
ColorView I • Format tab
Left, Right, Top, The fields entitled Left, Right, Top and Bottom show the number of pixels
Bottom that are to be clipped from each image edge. Each individual value can
range from 0 (no clipping) to almost the entire image in each respective
direction (complete clipping). If a value for the clipping has already been set,
the setting possibilities for clipping the opposite edge reduce themselves
respectively.
Interactive Click the Interactive button to be able to place the clipping interactively in the
image document. Move the pointer with the mouse button depressed to
change the size of the area to be acquired. The currently acquired area will
be indicated by a red frame. Move the mouse to alter the position of the
marked area within the image document. Rightclick once to fix the area to
be acquired.
No Clipping Click the No Clipping button to return the values for Left, Right, Top and
Bottom to 0, thus disabling Clipping.
48
Appendix I: Microscope settings
ColorView I
Appendix I: Microscope settings

Your digital camera can only achieve high image quality if the microscope
has been optimally set. A subsequent correction to the software can never
correct deficiencies in the image resulting from faulty settings made to the
camera and microscope.
The optical system microscope offers numerous setting possibilities. In
order to attain best acquisition results, the settings must be made precisely.
In spite of varying models and observation possibilities, the microscope's
image producing components are in accordance with each other.
A microscope basically consists of the optical components: projection objec-
tive and magnifier. The projection objective is the objective with which the
observation is to be carried out. The eyepiece takes on the function of the
magnifying glass. An additionally important component is the illuminator.
This illustration clarifies the
basic assembly of the
camera
Kamera
components of a micro-
scope in transmitted-light Lupe
Lens
mode.
Okular
ocular
objective Projection
Projektion
Objektiv
specimen
Probe
Kondensor
condenser
Beleuchtung
Illumination
Lampe
lamp
49
Illumination
Light is a decisive medium for creating images with a microscope. As a result, the light, i.e. illumination,
plays an important role in creating the image in the microscope.
The goal of the illumination settings is the equal distribution of light on the
sample with parallel light rays falling on the object. The illumination must be
bright enough to make all of the image details visible; however over-illumi-
nation leads to images being too bright and low-contrast. In addition, the
color of the light has a strong impact on the sample's color rendering.
Lamp Selection The tungsten-halogen lamps and gas discharge lamps are the most
commonly used lamps in microscopy. The light creating processes of both
lamp types are fundamentally different.
Tungsten-halogen Tungsten-halogen lamps comply with their functionality to a conventional
lamps light-bulb. Electricity flowing through a metal wire heats the wire to the extent
that light is produced (thermal emitter). In doing so, the wire emits a contin-
uous spectrum, hence a broad spectral band is emitted. One can control the
ColorView I • Illumination
brightness as well as the color (color temperature) of the lamp by controlling

the electrical current. Tungsten-halogen lamps can be widely used in
microscopy, especially when acquiring color images.
Gas discharge lamp Gas discharge lamps stimulate gas at the atomic level using strong electrical
fields. The excited atoms return to their initial state and, in doing so, give off
the energy which initially charged them in the form of electromagnetic radi-
ation: light. The resulting photons each have characteristic energy which is
typical for the gas being used. As a result, this type of lamp does not emit a
broadband spectrum like the halogen lamp, but instead emits discrete wave-
lengths. This fundamental physical process cannot be externally influenced;
hence, such a lamp's perceived color is independent of the operation condi-
tions. Because of this discrete spectrum, gas discharge lamps do not emit a
color-neutral illumination. Its adjustable high intensity light and the emission
of short-wave spectral bands required for fluorescent microscopy is an
advantage.
ND filter Many microscopes can insert ND filters into the light path, thus reducing the
intensity of the lamp's light. These ND filters ensure a uniform reduction of
light intensity throughout the entire frequency range without altering the
color temperature.
Köhler Illumination
This illumination process which was described by Dr. August Köhler as early
as 1893, is still an indispensable prerequisite for successful analysis using
light microscopy. The Köhler illumination enables an optimal, homogenous
illumination of solely the part of the sample to be analyzed.
Condenser The condenser carries the light onto the sample. The main components are
the aperture stop with which the size of the illuminated area is set and the
condenser lens which is responsible for the parallel emitted light rays. The
condenser can be opened and closed. The condenser is height-adjustable
and horizontally adjustable.
Note Set the aperture stop to 2/3 of the entire opening and do not use it to reduce
the brightness. To do that, you should always use ND filters or a lower lamp
voltage.
50
Principle of Köhler Illumi- will find comprehensive descriptions about the principle of Köhler illumina-
nation tion in microscopy text books. We will only give you a brief overview about
some important settings you should know.
Focussing the Field One vital step when setting up the Köhler illumination is the sharp view of
Diaphragm the field diaphragm at the specimen level.
1) Focus the object to be viewed.
2) Close the field diaphragm so that only a small bright spot is visible in
the eyepiece.
3) Focus the image in the field diaphragm by using the condenser's height
adjustment.
Adjusting the Now the illuminated area is optimally set.
Condenser 1) Open the field diaphragm so that its edge is still completely within the
visible area.
2) Center the image of the field diaphragm using the condenser's adjust-
ment screws.
" A precisely adjusted condenser guarantees that only the area of the
specimen to be viewed is illuminated.
Objective, Total Magnification and Useful Magnification
3) Repeat steps 1) and 2), until the edge of the field diaphragm is no
longer visible in the visual field.
Adjusting the Lamp You can generally skip adjusting the lamp with modern microscopes,
provided that the lamp is located in a frame and the bulb is in a centered
socket.
Objective, Total Magnification and Useful Magnification

There are no general rules for which objective you select; one should always
keep the specimen and goal of the observation in mind.
Terminology The two most important parameters of an objective (magnification and
numerical aperture) are engraved in the objective. First the magnification
and then the numerical aperture which is separated by a diagonal slash.
Hence, the code 40 / 0,65 marks an objective with a magnification of 40X
and the numerical aperture of 0.65.
Magnification An objective's engraved magnification is considerably important for the total
magnification. However, one should note that the range of useful magnifica-
tion should not be surpassed.
Numerical Aperture The numerical aperture is the characteristic of an objective which deter-
mines its capacity. The numerical aperture can also be found engraved into
the objective. It is based on the refraction index of the lenses being used as
well as the objective's angular aperture. The manufacturers are narrowly
limited when dealing with the refraction index, whereas the objective's
angular aperture can be increased by using appropriate measures. To
achieve this, a fluid with a high refraction index is placed between the cover
slip and the objective. These immersion objectives are accordingly marked
and should only be used with the appropriate immersion fluid.
Total magnification The system's total magnification, to which an installed TV adaptor contrib-
utes, is based on the actual size of the final image. This can be the image
on the monitor, as well as the image printed on paper. To determine the total
magnification, it is best to use a stage micrometer which can be shown on
the monitor or the printout. You can determine the total magnification by
dividing the length of the object shown by the set length on the micrometer.
Useful magnification The term Useful Magnification describes a total magnification of the opitical
51
system, using the resolution to the fullest without harming contrast and
sharpness. The useful magnification of the optical systems microscope
depends on the numerical aperture being used ( engraved on the objective).
The useful magnification is in the range of 500x to 1,000x the numerical
aperture.
Empty magnification A loss of contrast and sharpness is the result if total magnification is larger
than the useful magnification. Check to see if replacing the TV adapter with
a 0,5 or 0,63 model sufficiently reduces the total magnification. If this is not
possible, you can also use an objective with the same magnification, but
with a higher numerical aperture.
Monitor Settings for White Balance

Monitor Settings The monitor settings are decisive for the image you see on the monitor. Set
your monitor to the appropriate color temperature, should your device allow
this. To do this, compare the image's color impression on the monitor and in
the eyepiece.
ColorView I • Appendix II: How to install the camera
Note The color temperature setting can be altered on most monitors via the
monitor menu buttons located on the monitor. In other cases, the color
temperature can be altered via the operating system.
This is how you change the color temperature using the oper-
ating system:
1) Right click on Desktop.
" The Desktop's context menu is opened.
2) Select the entry Properties from the context menu.
" The Display Properties dialog box is opened.
3) Select the Settings tab.
4) Click the Advanced button.
" Another Display Properties dialog box is opened. The appearance
and function of this dialog box depends on the graphics card
installed on your system and your monitor
" Search for a tab with the name Color or Color Management.
Note The appearance of the operating elements might differ from the one being
described here because the adaptation of the color temperature is not part
of the operating system, but rather is made accessible by the monitor manu-
facturer or the graphics card being used.
5) Select a color temperature whose color impression best corresponds
to that seen in the eyepiece.
" Should your system offer a slide control with theoptions warm and
cold, do not alter your color temperature with these.
Appendix II: How to install the camera

Warning Please read the following carefully before attempting installation of
camera.
Requirements
52
The ColorView I can be used under the operating systems Windows 2000
and Windows XP. The installation procedure is described separately for
these two operating systems.
Installation sequence In order to avoid problems, you’ll have to follow the steps of the procedure
in the order given here.
Warning Install the image-analysis software first before connecting the camera
Windows 2000 to the FireWire board.
1) Check whether your computer system has a FireWire interface

onboard or a FireWire board installed.
2) Install the FireWire board provided with the camera.
3) Install the image-analysis software on your computer.
4) Install the camera and connect it to the computer system.
What if my computer If your computer system already has a FireWire interface onboard or a sepa-
system already has a rate FireWire board installed, you do not need to install the board provided
FireWire interface? with the camera. If you are not sure whether a FireWire interface is available,
open the Windows Device Manager via Start > Settings > Control Panel >
System > Hardware > Device Manager....
The FireWire board IEEE-1394
An entry called ’IEEE 1394 Bus host controllers’ shows that the needed
FireWire interface is available.
General Warnings!
• CCD image sensors are easily damaged by static discharge. Do not
disassemble the camera housing.
• Do not attempt to touch the internals of the camera with your bare
hands.
• Do not touch cover slip (over the CCD chip) with your hand or have any
object come in contact with glass surface. Should dust stick to the
cover slip, blow it off gently with an air blower. (For dust stuck due to
static electricity, ionized air is recommended.)
•
53
It is best to use a LensPen to clean the cover slip. Do this only when
absolutely necessary.
• Do not expose to strong light (sunlight) for long periods.
• Exposure to high temperature or humidity will affect the characteristics
of the camera. Avoid storage or usage under such conditions.
• Do not subject the camera to too much mechanical shock.
• The FireWire cable plug is not symmetrical. You must make sure that
the cable plug is facing the same way as the camera socket when you
plug the cable into the camera jack.
• Do not disconnect the FireWire cable while the image-analysis soft-
ware is running.
The FireWire board IEEE-1394

Warning! • The components of these devices are easily damaged by static
discharge. Wear grounding wrist straps when handling them.
• Do not plug in or unplug the cards from the PCI slot while the PC is on.
ColorView I • Installation Procedure - Windows 2000
• Please follow the installation manual in this booklet step by step to

avoid any incorrect connections. Incorrect connections may result in
serious damage to your PC system and/or the camera.
Installation Procedure - Windows 2000

Installation of the FireWire board IEEE-1394
When is this necessary? This procedure is necessary only if the PC does not have a FireWire inter-
face on-board hardware configuration or if it is not yet equipped with a
FireWire board.
The FireWire board
How to install the 1) Turn off the PC and disconnect the power supply.
FireWire board 2) Remove the cover from the PC.
3) One PCI slot is required for the FireWire board. Insert the board into the
slot.
FireWire board installed
54
4) Replace the cover onto the PC.

Installation of the image-analysis software
5) You are now ready to first install the image-analysis software and then
the camera.

Warning! The image-analysis software must be installed first before connecting the
camera to the FireWire board.
Requirements, PC The ColorView I camera requires the image-analysis software, (3.2 version),
and Microsoft Windows 2000 (or later).
camera driver installation Installation of the SIS View FireWire camera driver is simply done via the
image-analysis software setup. You are only required to select the camera
during setup.
Installation of the image- For installation, follow the instructions given on the monitor. You do not have
analysis software to select a digitizer in the Input Device (Digitizer) Selection dialog box.
Select Olympus Imaging
Solutions as manufacturer
and ColorView I as camera.
55
Precalibration You will be advised to carry out a precalibration of the camera when
installing the image-analysis software.
In this dialog box you can

activate the precalibration.
Select the TV adaptor's magnification from the list which has the same
name. Should you not be working with a remote microscope, clear the check
box. Then, mark the checkbox for all mounted objectives in the Magnifica-
tion of the microscope objectives list.
Note Should you use objectives with a magnification that does not appear in the
list, a manual calibration must be carried out for these objectives. A guide is
part of the image analysis system's step by step guide.
Note The image-analysis software can support several same or different types of
FireWire cameras simultaneously. You differentiate Color View I cameras
via their serial numbers when creating a new logical input channel via Image
> Set Input... > New Channel. A camera’s serial number is located directly
below the FireWire jack on the camera housing.
56
The camera’s serial number

is located below the
FireWire jack.
Installation of the camera and cables
Installation of the camera and cables

Don’t forget... The image-analysis software must be installed first before connecting the
camera to the FireWire board.
How to install the camera 1) The diagram shows a typical installation of a light microscope. You
should have all required components ready at hand.
FireWire cable
ColorView I
C-mount adaptor
PC with image analysis software

and FireWire port
57
FireWire plug
2) Remove the protective cap from the camera.

3) Firmly screw the TV adaptor (0,5x or bigger) onto the camera. This
should be done with the CCD-chip side of the camera facing down to
minimize the chance of dust landing on its surface.
4) Install this assembly onto the microscope's camera mount. Some
microscopes may require additional optical elements installed between
the camera mount and the C-mount adapter. You may also need to
align the orientation of the microscope with that of the stage, and also
match the focus of the camera image with that of the eyepiece image.
These settings can be done later when you are able to obtain an image
on the screen.
5) Plug one end of the FireWire cable into the camera connector.
" The FireWire cable plug is not symmetrical. You must make sure
that the cable plug is facing the same way as the camera socket
when you plug the cable into the camera jack. The camera chip
may be ruined otherwise.
Make sure the cable plug is
facing the right way before
you plug it into the camera
socket
6) Plug the other end into one of the small connector sockets on the
FireWire board.
" There are two or more jacks depending on the FireWire board. You
can use any of them.
" The Digital Signature Not Found MS Windows dialog box(es)
appear on your screen. Confirm the message as often as neces-
sary until the dialog boxes disappear.
58
7) Open the Windows Device Manager via Start > Settings > Control
Panel > System > Hardware > Device Manager....
" The Device Manager dialog box contains the correct camera driver
entry Olympus Soft Imaging Solutions FireWire Camera 2.1.2 (Soft-
Hard). The camera is ready for use.
Troubleshooting
The Windows Device Manager shows the IEEE 1394 FireWire board and the camera
Olympus Soft Imaging Solutions FireWire Camera 2.1.2 (SoftHard).
Troubleshooting
What do I do if the An error message appears when starting the program, even though you
camera cannot be have completely installed the camera and the image-analysis software:
found?
59
The image-analysis software has not been successful in creating a connec-

tion to the camera. Confirm the message with OK.
Check to see if the connector cable between the camera and the computer
has been correctly plugged-in. Should the connecter be loose or not plugged
in correctly, correct this by firmly plugging them into the corresponding jack.
Restart the image-analysis software. Should the problem still exist, continue
with the following section.
What to do when the The error message No SIS-View FireWire camera has been found will also
camera has been appear when starting the program should the camera have been connected
connected before the before installing the image-analysis software. In this case, Windows 2000
image-analysis software has already installed its own camera driver, which prevents the error-free
installation
communication between the camera and the computer. Installing the image
analysis software now does not install the correct camera driver since this is
forbidden by Windows 2000.
If the camera is hooked up to the computer, you have to install the correct
camera driver from the software’s setup CD separately from the software
installation. Do the following:
1) Select the Start > Settings > Control Panel > System > Hardware >
Hardware Wizard... command.
2) Click on the Next button.

60
Troubleshooting
3) Select the Add/Troubleshoot a device option and click on the Next

button.
61
4) Select the entry Olympus Soft Imaging Solutions FireWire Camera

2.1.2 (SoftHard Technology) Devices list and click on the Next button.
5) Click on the Finish button.

62

Troubleshooting
7) Select the Search for a suitable driver for my device (recommended)

option and click on the Next button.
63
8) Insert the image-analysis software setup CD into your computer’s CD-

ROM drive. Minimize the appearing setup dialog box.
9) Make sure that only the CD-ROM drives check box is selected and click
on the Next button.
" Wait until Windows has found the correct camera driver. The
driver's data file is mm282osis.inf. You will find it on the CD-ROM
in Directory\program\support\cameras\sis_firewire\drivers\win2k.

" The Digital Signature Not Found dialog box appears on your
screen.
64
11) Click Yes.

" Wait until the camera driver has been installed.
Troubleshooting
12) Click on the Finish button.

" The Digital Signature Not Found MS Windows dialog box(es)
appear on your screen. Confirm the message as often as neces-
sary until the dialog boxes disappear.
65
13) Open the Windows Device Manager via Start > Settings > Control
Panel > System > Hardware > Device Manager.... The Device Manager
dialog box The Device Manager dialog box contains the correct camera
driver entry Olympus Soft Imaging Solutions FireWire Camera 2.1.2
(SoftHard).
ColorView I • Installation Procedure - Windows XP
The Windows Device Manager shows the IEEE 1394 FireWire board and the camera driver
Olympus Soft Imaging Solutions FireWire Camera 2.1.2 (Softhard).
14) Install the image-analysis software. During the installation check the
Firewire check box in the Input Device (Digitizer) Selection dialog box.
Mark the ColorView I in the Camera SelectionI dialog box.
Installation Procedure - Windows XP

66
Installation of the FireWire board IEEE-1394

When is this necessary? This procedure is necessary only if the PC does not have a FireWire inter-
face on-board hardware configuration or if it is not yet equipped with a
FireWire board.
The FireWire board
How to install the 1) Turn off the PC and disconnect the power supply.
FireWire board 2) Remove the cover from the PC.
3) One PCI slot is required for the FireWire board. Insert the board into the
slot.
FireWire board installed
67
4) Replace the cover onto the PC.

5) You are now ready to first install the image-analysis software and then
the camera.

Warning Install the image-analysis software first before connecting the camera to the
FireWire interface.
Requirements, PC The Color View I camera requires the 823 image-analysis software and
Microsoft Windows XP (or higher). The image-analysis software also
supports multithreading on multiprocessors, thus making full use of the
power of the latest multi-CPU PC’s.
camera driver installation Installation of the SIS View FireWire camera driver is simply done via the
image-analysis software setup. You are required to select FireWire and
specify the additional camera driver during setup. Then you have to get
Windows to use the correct camera driver.
Installation of the image- For installation, follow the instructions given on the monitor. You do not have
analysis software to select a digitizer in the Input Device (Digitizer) Selection dialog box.
Select Olympus Imaging
Solutions as manufacturer
and ColorView I as camera.
68
Related Topics
Installation of the camera driverS. 72
Precalibration You will be advised to carry out a precalibration of the camera when
installing the image-analysis software.
In this dialog box you can
activate the precalibration.
Select the TV adaptor's magnification from the list which has the same
name. Should you not be working with a remote microscope, clear the check
box. Then, mark the checkbox for all mounted objectives in the Magnifica-
tion of the microscope objectives list.
Note Should you use objectives with a magnification that does not appear in the
list, a manual calibration must be carried out for these objectives. A guide is
part of the image analysis system's step by step guide.
Note The image-analysis software can support several same or different types of
FireWire cameras simultaneously. You differentiate ColorView I cameras
via their serial numbers when creating a new logical input channel via Image
69
> Set Input... > New Channel. A camera’s serial number is located directly
below the FireWire jack on the camera housing.
The camera’s serial number

is located below the
FireWire jack.
How to install the camera 1) The diagram shows a typical installation of a light microscope. You
should have all required components ready at hand.
FireWire cable
ColorView I
70
C-mount adaptor
PC with image analysis software

and FireWire port
FireWire plug
2) Remove the protective cap from the camera.

3) Firmly screw the TV adaptor (1/1,8" or bigger) onto the camera. This
should be done with the CCD-chip side of the camera facing down to
minimize the chance of dust landing on its surface.
4) Install this assembly onto the microscope's camera mount. Some
microscopes may require additional optical elements installed between
the camera mount and the C-mount adapter. You may also need to
align the orientation of the microscope with that of the stage, and also
match the focus of the camera image with that of the eyepiece image.
These settings can be done later when you are able to obtain an image
on the screen.
5) Plug one end of the FireWire cable into the camera connector.
" The FireWire cable plug is not symmetrical. You must make sure
that the cable plug is facing the same way as the camera socket
when you plug the cable into the camera jack. The camera chip
may be ruined otherwise.
Make sure the cable plug is
facing the right way before
you plug it into the camera
socket
6) Plug the other end into one of the small connector sockets on the
FireWire board.
" There are two or more jacks depending on the FireWire board. You
can use any of them.
7) Continue with the camera driver installation before starting the image-
71
analysis software.
Installation of the camera driver

The SIS View FireWire camera driver is not signed and therefore not used
automatically under Windows XP. After having installed the image-analysis
software and connected the camera to the computer - or vice versa - you
have to go through the following steps to get Windows XP to use the correct-
camera driver:
1) Open the Windows Device Manager via Start > My Computer > (right
mouse) > Properties > Hardware > Device Manager....
72
" The Device Manager dialog box shows the Generic 1394 Desktop
Camera entry in the Imaging devices section. The exclamation
mark indicates a device conflict.
2) Select the Generic 1394 Desktop Camera entry and click on the
Update driver button in the device manager button bar.
3) Select the Install from list or specific location (advanced user) option
and click the Next button.
73
4) Select the Don’t search. I will choose the driver to install. option and
click on the Next button.
" The Model may already contain one ore more entries based on
previous software installations.
5) Select the entry Olympus Soft Imaging Solutions FireWire Camera
2.1.2 (SoftHard) or higher.
" If this entry is missing, click on the Cancel button. Repeat the instal-
lation of the image analysis program including the selection of
ColorView I as camera.
" Do not select an entry reading OLYMPUS CORPORATION
FireWire Camera.
" The next Hardware Update Wizard dialog box will be opened in the
background displaying the message to wait while the Wizard
74
installs the software.

" Additionally, the Hardware Installation dialog box will pop up in the
foreground.
Related Topics
Installation of the image-analysis softwareS. 67
7) Click on the Continue Anyway button.

" The Hardware Update Wizard dialog box which had been in the
background, will become visible.
75
" After having finished the camera driver installation the following
dialog box will be opened.
8) Click the Finish button to close the Hardware Update Wizard.

" In the Device Manager dialog box there is still the same Generic
1394 Desktop Camera entry but the exclamation mark indicating
the device conflict should have disappeared. The camera is now
ready for use.
76
In case of problems with the camera...
In case of problems with the camera...

Should you have questions or should problems arise which you cannot solve
with the help of the documentation provided, please feel free to contact our
customer service - preferably via e-mail. Our customer-service personnel
will be more than happy to assist you.
1) Please clarify before hand when and under what circumstances the
problem arose.
• It would be best if you can clearly reproduce the problem/error. You
will therefore make it easier for you and our customer-service
personnel to solve the problem.
2) Please write down the exact wording of the (error) messages which
appear.
3) As an alternative, you can make "Snapshots" of these messages. You
can make a snapshot of your active window by using the key combina-
tion [Alt+Print]. A copy of the active window is stored to your clipboard
and can be easily attached to an e-mail using [Ctrl+V].
4) Since very large e-mails can create transmission problems both for the
sender and the receiver, do not copy "screenshots" of the entire
contents of the screen. For the same reasons it is advisable to reduce
the color depth of the snapshots.
5) Should the camera basically work, but problems with image quality
arise, save an image in which the problems are clearly visible in TIF
format. Insert this example image in your e-mail to our customer
service.
6) Start the image-analysis software.
7) Select the ? > Info... command to open the Info dialog box.
" The Info-Dialog box shows you information about the software
package, the Build number and the serial number of your image
analysis program as well as the operating system being used.
• The software data mentioned should be at hand when contacting
our customer service personnel.
8) Please write an e-mail clearly describing the problem (including snap-
shots
77
and example images), as well as the software data from the Info dialog
box to our customer service address:
support.osis@olympus-sis.com
• The quickest and most convenient way is to use the automatic

email settings for your customer service query:
By using the ? > Info... > System Info... > Send button, an automatic
e-mail is created which you can then complete and send to us. If
you have made an example in TIF format, please attach this to the
e-mail as well. Before sending, read the notes in your e-mail box the
data about your system which you will send to us in this e-mail.
Should you not be able to send e-mails from your computer, use
the? > Info... > System Info... > Info save save button to save the
files and to send them from another computer.
• You may of course contact us via telephone or fax:
Tel.: (+ 49) 2 51 / 7 98 00-444
Fax: (+ 49) 2 51 / 7 98 00-6060
• WEEE DECLARATION
WEEE DECLARATION
Waste Electrical and In accordance with European Directive 2002/96/EC on Waste Electrical and
Electronic Equipment Electronic Equipment, this symbol indicates that the product must not be dis-
posed of as unsorted municipal waste, but should be collected separately.
Refer to your local distributor for return.
78
CE DECLARATION OF CONFORMITY
Product Type CCD Camera

Brand Name Soft Imaging System
Model Designation Color View I
Name and Address of manufacturer or representative
Manufacturer Soft Imaging System GmbH

Johann-Krane-Weg 39
D-48147 Münster
The product complies with the requirements of the following European directives:
89/336/EWG Directive of the council on the approximation of the laws of Member States
relating to electromagnetic compatibility changed by directive 91/263/ECC, 92/
31/ECC and 93/68/ECC of the council.
Compliance was proved by the application of the following electromagnetic
compatibility standards:
EN 55022 : 1998 + A12000, cl. B
EN 55024 : 1998
EN 61326 : 1997 + A1 : 1998 + A2 : 2001 / appendix A
EN 61000-6-2 : 2001
EN 61000-6-3 : 2001
EN 61000-4-2 : 1995 + A1 : 1998 + A2 : 2001
EN 61000-4-3 : 1995
EN 61000-4-4 : 1995 + A1 : 2001 + A2 : 2001
EN 61000-4-6 : 1996 + A1 : 2001
Date: November 4th, 2003
Dr Manfred Schierjott November 4th, 2002

Date
Index
Index
E
Enabling Shading Correction 27
A Exposure time
analySIS, version 4 Input 41
Automatic Gain Display Exposure time, intX 10
Input 43
F
B Firmware Version 42
Binning 47 Fluorescent microscopy
Black & white mode Black balance 22, 30
Gamma Correction 30
Black balance 22, 30
G
Gamma Correction
C Black & white mode 30
Camera configuration 39 RGB mode 19
Camera Control
Black balance 22, 30
Color Settings 19 H
Partial Readout 22
Sharpen filter 18 Housing temperature 42
Sharpness Monitor 20
Camera resolution
In input channel 41 I
Camera Type 42 Image scaling, Display tab 46
Clipping 47 intX
Color Settings 19 Calibration 10
Gamma Correction 19 Online shading correction 29
Saturation 20
80
Color temperature of the monitor 52

ColorView III Information 41
Configure Input 40 K
Köhler illumination, microscope 50
D
Device ID 42 L
Device Information 41 Live overlay 45
Display tab
Image scaling 46
Over exposure 43 M
Palette 47
DLL version, ColorView III information 42 Microscope 49
Duplicate Channels 40 Empty magnification 51
Illumination 49
Numerical Aperture 51
Objective 51
Index
Total magnification 51 Sharpness Monitor 20

Useful magnification 51 Special 17
Microscope settings 49 System Info 77
Mirror 41
Monitor Settings 51, 52
T
Tab Format
N Binning 47
NetCam 34 Clipping 47
in the internet 36 Image format 47
In the intranet 35 Tab Input
Settings 37 Automatic gain display 43
New Channel 39 Exposure time 41
Info 41
Mirror 41
O Resolution 41
Sharpen filter 41
Online histogram 44 Temperature 42
Online shading correction 23 Tools 33
intX 29
Over exposure 43
Over Exposure Warning 43
U
Useful magnification, Microscope 51
P
Partial Readout 22
PC, Requirements 4 W
Web Server 35
White balance 17
R Execute 17
Real time FFT 45
81
Requirements, PC 4
Resolution
Input 41
RTFFT
Control 32
Display 33
S
Select device 40
Serial Number 77
Serial number 42
Set Input 39
Shading Correction
Activate 27
Sharpen filter 18
Sharpen filter Activate 41

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Manual

© Olympus Soft Imaging Solutions GmbH

The ColorView I - camera 4

WHEN CAN COLORS APPEAR TO BE FALSE? .....................6

Acquiring images with manual controls 11

FURTHER OPTIONS IN THE CAMERA CONTROL ..............17

Online shading correction 23

USING THE SHADING CORRECTION ..................................24

Using the black & white mode 30

ACQUISITIONS WITH REAL TIME FFT .................................31

INSTALLING THE WEB SERVER .......................................... 35

Setting up the camera 39

LOGICAL INPUT CHANNELS ................................................ 39

Appendix I: Microscope settings 49

Appendix II: How to install the camera 52

INSTALLATION PROCEDURE - WINDOWS 2000 ................ 54

In case of problems with the camera... 77

The ColorView I - camera

Exposure times 100 µs to 160 s

Hard- / Software Required

Overview of the chapters

"Appendix II: How to install the camera" contains detailed instructions on

2) Click the Acquire > button.

When can colors appear to be false?

When is it necessary to perform a white balance?

This is how you perform a white balance with Intelligent Expo-

When can the image be too dark?

In the default setting,

2) Click the Set ROI for White Balance button.

With the default setting of

2) Move the Exposure adjustment slide control to the right.

Calibrating Intelligent Exposure

The second calibration step consists of measuring the signals at various

Exposure Time Lock

button anew to return to the dynamic adjustment of the exposure time.

Acquiring images with manual controls

Preparing the input channel

8) Click the Snapshot button to acquire a snapshot with the current

Automatic Gain Display and fixed scaling

4) Position the ROI in such a way as to encompass the area that is to be

Difference between Automatic Gain Display and fixed scaling

Button Area Effect

Further options in the Camera Control

Performing a white balance with manual acquisition

If you start a live acquisition with an activated Sharpness Monitor or activate

Using the sharpness monitor

Using the Partial Readout

" Fix the area with a right click

Using the black balance

Online shading correction

Using the Shading Correction

How the Shading Correction functions

Acquiring Reference Images

Enabling Shading Correction

Shading correction when working in black & white

Repeat or Update the Reference Acquisitions

Use of the Online Shading Correction

intX and Online Shading Correction

Using the black & white mode

Performing a black balance

Acquisitions with Real time FFT

forms). The Fourier image is continually updated during a live acquisition. It

The Fourier Image of a