Raman Users Training

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2007
2009 HORIBA,
HORIBA, Ltd.Ltd.
All All rights
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 Map treatement

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2009 HORIBA,
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Defining the map Defining the map
The easiest method is to define the size of the map in the video window. On the left
side of the screen there is a tool bar which allows define the rectangular map, line
maps, circle map or random points.

Then we can open the Mapping properties window (use the icon ) to adjust other
parameters like step or the matrix size. Not only maps XY can be defined. But the
depth profiles (motor Z required), or temperature (with the stage Linkam), or kinetics
(see next slides), as well. To add a new line, click-right and choose Insert, the window
with all possible map types will appear.

2007
2009 HORIBA,
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Defining the map Defining the map
If you want to define XY map bigger than the surface seen in the Video window, close the Video window and define the
size of the map directly in the Mapping Properties window. Do not forget to activate Relative option (last column). In this
way the LabSpec will take the green point as the (0, 0) point. If the option Relative is not activated, the values in From and
To will be taken as the absolute values.

Without the option Relative With the option Relative

The position of the laser spot
The position of the laser spot

0
0

table
table
0
0

Map: X de -5 +5m Map: X de -5 +5m

Y de -5 +5m Y de -5 +5m

2007
2009 HORIBA,
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Defining the map Defining the map
Kinetics is special type of the mapping which allows to do repeat measurements. To define it:
1. Choose Time in the Mapping properties.
2. Define the beginning time (From), the end time (To) and the delay between the BEGINNINGS of
each measurement (Increment).
3. Define the acquisition time in the bottom bar in the acquisition part. The total acquisition time
(number of acquisitions x acquisition time) should be smaller or equal the delay defined in
Increment.

2007
2009 HORIBA,
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Data saving Data saving
Mapping
The file *.ngc contents three windows: SpIm, Point and Map.

to save a mapping, activate SpIm window

video image should be saved separately (*.ngv file)

each window can be saved separately: activate a chosen window

all the spectra of the mapping can be saved separately: chose the option menu
File Split

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2009 HORIBA,
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Mapping treatment

Mapping and depth profile analysis

Map Raman
SpIm image based
contents all on the band
the spectra of position
the mapping
or depth
profile

Point
spectrum
corresponding
to the cursor
position

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2009 HORIBA,
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Mapping treatment

Mapping analysis using R V B cursors

Activate Use to show the

image corresponding to the
From and To can be sued
cursor
to define the cursors
postions
Activate Baseline to take
into account the baseline

Activate Green/blue to
generate image
corresponding to the ratio
of the green map to the Correct accepts the
blue map changes done in the
Point window
Activate Spectrum to open
the Point window

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2009 HORIBA,
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Mapping treatment

Mapping analysis with peak fitting

507.8
514.8
-30 -30
507.6 514.6

-20 -20 514.4

507.4

Y (m)

Y (m)
-10 -10 514.2
507.2
514.0
0 0
4 m 507.0 4 m 513.8

10 10 513.6
-20 0 -20 0
X (m) X (m)

507.8
514.8
-30 -30
507.6 514.6
-20 -20
507.4 514.4
Y (m)

Y (m)
-10 -10 514.2
507.2
514.0
0 507.0 0
4 m 4 m 513.8

10 10 513.6
-20 0 -20 0
X (m) X (m)

2007
2009 HORIBA,
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Mapping treatment

Normalisation and spectra correction

Main functions :
normalisation to a chosen peak (or a part of the spectrum)
zero shifts all the spectra to the zero level
mathematical functions permit to apply a mathematical operation to all the spectra
(i.e.: subtract a spectrum from all the spectra of the mapping)
In case of the normalisation, it is
necessary un general to define the
spectral region using cursors or in the
field Limits:

In case of zero level, if all the

spectrum should be considered, the
field Limits has to deactivated.

2007
2009 HORIBA,
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Mapping treatment
Example of normalisation : analysis of the skin
hydration level
The spectra were normalised to the protein characteristic After normalisation
4.5

2942.0
band (stretching C-H) which is unchanged in the skin 4.0

3.5

3.0 Stretching C-H

Intensit (coups)
2.5

2.0

1.5

1.0 Stretching O-H

0.5
Intensit (coups)

4 000 0.0
0
2 800 3 000 3 200 3 400 3 600
Shift Raman (cm-1)
2 000 20 )
m
Z(

40
1 000 2 000 3 000
Shift Raman (1/cm) depth

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Mapping treatment

Modelling

To :

- analyse the mapping results in the

automatic way (without cursors R, V, B)
using the reference spectra (factor
analysis). In this case the analysis is based
on the whole spectrum

- to show the distribution of the components

in case if they are more than 3

The reference spectra can be :

- Generated by the software

- Given by the user

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2009 HORIBA,
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Mapping treatment

Modeling : Analyse without reference spectra

For :
- quick qualitative anaysis,
- determine the number of the components

Parameters :
- componenets or factors : the number of expected
components. The factors corresponding to the components
are determined using the classification method.
Advice : The
normalisation get rid off - treshold (%) : allows to exclude the spectra of a very low
the surface effects intensities

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2009 HORIBA,
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Mapping treatment

Modeling : Analyse without reference spectra

Spectra created by LabSpec5

The result (score) of the decomposition

Experimental spectra to the factors (Direct Classical Least
Squares)

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Mapping treatment
Modeling : Analyse with the reference spectra

To :
- Improve the qualitative analysis : better ration S/N of
the Raman image or profile (based on the whole
spectrum and not one band)

- In some case the QUANTITATIVE analysis is

possible
In this case its better that all the reference
spectra are saved in the same conditions =>
the intensities can be used directly to determine
the concentration WITHOUT the normalisation
(in this case the normalisation means the loss
of the information)
The reference spectra :

- can be the recorded on the pure components

- can be extracted from the mapping from the
zones where the components are well
separated

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Mapping treatment
Modelling : Analyse with the reference spectra
examples
Analyse with cursors

4 000

3 500 800

3 000

Intensity (cnt)
600
2 500
Intensity (cnt)

Adhesive Polymer
2 000
400
1 500

1 000 200

500

0
0
20 0 -20
800 1 000
Raman Shift (cm
1 200
-1
)
1 400
Piezo (m)

Analyse using modelling

8 000 _1
_2
1.0 7 000

6 000
0.8
Intensity (cnt)

Intensity (cnt)
5 000
0.6 Adhesive Polymer
4 000

0.4 3 000

2 000
0.2
1 000
0.0
0
2 500 3 000 3 500
-1 20 0 -20
Raman Shift (cm )
Piezo (m)

2007
2009 HORIBA,
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 3D Image
Three options in the pop-up menu and than
(rigth-click):

Firstly

and more

2007
2009 HORIBA,
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reserved.
 www.horiba.com/scientific

2007 HORIBA, Ltd. All rights reserved.

2007
2009 HORIBA,
HORIBA, Ltd.Ltd.
All All rights
rights reserved.
reserved.