Microscope

18th century- Technical

innovations improved microscopes
leading to microscopy becoming
popular among scientists. Lenses
combining two types of glass
reduced the "chromatic effect" the
disturbing halos resulting from
differences in reflection of light
1674- Anton van Leeuwenhoek
built a simple microscope with
only one les to examine blood,
yeast, insects and many other tiny
objects. Leeuwenhoek was the first
person to describe bacteria, and he
invented new methods for grinding
and polishing microscope lenses
that allowed for curvatures
providing magnification of up to
270 diameters, the best available
lenses at that time.
1665- English physicist, Robert
Hooke looked at a sliver of cork
through a microscope lens and
noticed some pores or cells in it.
1590- Two Dutch eye glass makers,
Zaccharias Janssen and son Hans
Janssen experimented with
multiple lenses placed in a tube.
The Janssens observed that viewed
objects in front of the tube
appeared greatly enlarged, creating
both the forerunner of the
compound microscope and the
telescope.
Circa 1284-Italian Salvino D'
Almante is credited with inventing
the first wearable eye glasses
Circa 1000AD- the first vision and
was invented (inventor unknown)
called a reading stone. It was a
glass sphere hat magnified when
laid on top of the reading materials.

Using High Power- Focus only

with fin focus. Hopefully, the
specimen will come into focus
easily. Do not change focus
dramatically
Rotate to 40x objective , locate
desired portion of specimen in the
center of the field. Refocus very
carefully so that the specimen is
focused as sharply as possible. (Do
not alter focus for the following
steps)
Partially rotate so that 40x and
100x objectives straddle the
specimen.
Rotate so that the 100x oil
immersion objective touches the oil
and clicks into place
Place a small drop of oil on the
slide in the center of the lighted
area. (Take care not to dribble on
the stage) Put the small drop of oil
directly over the area of the
specimen to be examined.
Clean Up!: When you have
finished for the day, wipe the 100x
oil immersion objective carefully
with lens paper to remove all oil.
Wipe oil from the slide thoroughly
with a Kimwipe. Cleanse stage
should any oil have spilled on it.
Recap the immersion oil container
securely, replace in drawer.
Using the Microscope- The proper
way to focus a microscope is to
start with the lowest power
objective lens first and while
looking from the side, crank the
lens down as close to the specimen
as possible without touching it.
Now, look through the eyepiece
lens and focus upward only until
the image is sharp. If you can't get
it in focus, repeat the process again.

Once the image is sharp with the

low power lens, you should be able
to simply click in the next power
lens and do minor adjustments with
the focus knob. If your microscope
has a fine focus adjustment, turning
it a bit should be all that's
necessary. Continue with
subsequent objective lenses and
fine focus each time.
Light- makes the specimen easier
to see
Diaphragm- controls the amount of
light going through the specimen.
Many microscope have a rotating
disk under the stage. This
diaphragm has different sized holes
and is used to vary the intensity
and size of the cone of light that is
projected into the slide. There is no
set rule regarding which setting to
use for a particular power. Rather,
the setting is a function of the
transparency of the specimen, the
degree of contrast you desire and
the particular objective lens in use.
Stage Clips- hold the slides in
place. If your microscope has a
mechanical stage, you will be able
to move the slide around by turning
two knobs. One moves it left and
right, the other moves it up and
down.
Objective lenses- adds to the
magnification. Usually you will
find 3or 4 objective lenses on a
microscope. They almost always
consist of 4x, 10x, 40x and 100x
powers. When coupled with a 10x
(most common eyepiece lens), we
get total magnifications of 40x (4x
times 10x) , 100x, 400x, and
1000x. The shortest lens is the
lowest power, the longest one is the

lens with the greatest power.

Lenses are color coded. The high
power objective lenses are
retractable (i.e 40XR) This means
that if they hit a slide, the end of
the lens will push in (spring
loaded) thereby protecting the lens
and the slide.
Body tube- connects the eyepiece
to the objective lenses
Base- the bottom of the microscope
used for support
Fine Adjustment Knob- small,
round knob on the side of the
microscope used to fine-tune the
focus of your specimen after using
the coarse adjustment knob
Coarse Adjustment Knob- moves
stage (or body tube) up and down
Stage- the flat platform where you
place your slides
Arm- supports the tube and
connects it to the base
Microscope Care
- Always Carry with 2 hands
- Never touch the lenses with your
fingers
-Only use lens paper for cleaning
- Do not force knobs
-Keep objects clear of desk and
cords
-When you are finished with your
scope rotate nosepiece so that it's
on the low power objective, roll the
stage down to the lowest level,
rubber band the cord the replace
the dust cover.
Dissection Microscope- is light
illuminated. The image that
appears is three dimensional. It is
used for dissection to get a better
look at the larger specimen. You
cannot see individual cells because
it has a low magnification (also
called stereo microscope)

Scanning Electron Microscope

(SEM)-use electron illumination.
The image is seen in 3-D. It has
high magnification and high
resolution. The specimen is coated
in gold and the electrons bounce
off to give you an exterior view of
the specimen. The pictures are in
black and white.

Why does it matter?

Safe working protects: you, other

lab workers, cleaners, visitors and
your work
What does the law say? Health
Safety at Work etc Act 1974
- You must work safely
-You must not endanger others
-You must not misuse safety
equipment
Penalty- up to 2 year in prison and
or an unlimited fine
The Management of Health and
Safety at Work Regulations 1999
Control of Substances Hazardous
to Health Regulations 2004
You must perform Risk
Assessments

How to do a Risk Assessment?

Determine hazards and evaluate

risks
Use all relevant available data
Determine controls needed to
minimize those risks
Document the assessment
Agree with your supervisor
Use those control measures

Control Measures (in order of preference)

Use a less risky substance

Use a safer form of that substance

(eg solution instead of powder)
Totally enclose the process (eg a
glove box)
Partially enclose the process (eg
with a fume cupboard)
Ensure good general ventilation
Safe systems of work
Reduce exposure times, increase
distance, reduce volumes
Personal protective equipment (as a
last resort for primary protection)

Protecting Yourself

Wear the clothing and protective

wear identified in your risk
assessment
Laboratory coats must be kept
fastened
Don't wear sandals or open shoes
Long hair must be tied back
(gloves) There are many different
types of protective glove
Use the correct ones for the job you
will be doing
Remember that you need to select
chemical protection gloves
according to the materials and or
substances with which you will be
working
Remove your gloves before using
instruments, telephone, and leaving
the laboratory

Laboratory Hygiene

Never eat, drink, or smoke in a

laboratory
Never apply cosmetics
Never touch your face, mouth or
eyes
Never suck pens or chew pencils

Always wash your hands before

you leave and especially before
eating

What are the general hazards in a

laboratory?

Fire
Breakage of glassware
Sharps
Spillages
Pressure equipment and gas
cylinders
Extremes of heat and cold
Chemical Hazards
Biological Hazards
Radiation

Avoiding Fires

Flammable substances
-Use minimum quantity
-Store in special storage cabinet
-Use temperature-controlled
heating sources (eg. water-bath
rather than hot plate or Bunsen
Burner)

Minimize Fire Damage

Make sure corridor fire doors and

laboratory doors are kept shut at all
times

Fire Safety

Make sure that you know what to

do: if you have a fire, if you have a
fire alarm
If you are a staff member, you must
attend fire training annually. Post
graduates should also seriously
consider doing so.

Glassware

Use correct techniques for the

insertion of tubing onto glassware
Never use glassware under pressure
or vacuum unless it is designed for
the job and suitably shielded
Dispose of chipped or broken
glassware- it is a risk to you and
others
Always dispose of broken glass in
a glass bin or sharps bin and not in
a general waste bin

Spillages

Clear up spillage promptly

You will already have determined
how to do this as part of your risk
assessment
Dispose of any hazardous material
as toxic waste
Messy workers are usually poor
workers!

Electrical Equipment

Always do a visual check on

electrical equipment before use,
looking for obvious wear or defects
Never use defective equipment

General Tidiness

Keep you workplace tidy

Clear up waste, deal with washing
up and put things away as you
finish with them
Make sure everything is safe before
you leave things unattended
A tidy laboratory avoids accidents
to everyone

Laboratory Equipment

Never use any laboratory

equipment unless you are trained
and have been authorized to do so

As well as injuring yourself you

may cause very costly damage

First Aid

All laboratory workers should

undergo simple first aid training
- For all chemical splashes, wash
with plenty of water for 10 minutes
-Control bleeding with direct
pressure avoiding any foreign
bodies such as glass

Protecting your Health

If you have an allergy to lab

materials or suffer from a medical
condition which may affect you in
the laboratory (eg diabetes or
epilepsy) ensure that your
supervisor knows.

Waste Materials

Part of your risk assessment will be

to determine how to dispose of
waste lab materials safely
- Solvents and oils must be
segregated into the correct waste
bottle or drum
-Your department will help you
determine what to do with
chemical or biological materials
Do not put materials down the
drain or in with normal waste
unless authorized to do so

Working outside normal hours and at

weekends

You will need to attend training

courses and have permission from
your Head of Department before
working outside normal hours
Most experimental work is not
permitted

Your supervisor will explain the

requirements in more detail

When in doubt-ASK!

Do not carry out a new or

unfamiliar procedure until you
have been fully trained and
understand the precautions
necessary for safe working

Urinalysis Notes: (Important)- Typical

Substances Tested and Significance

pH- partial assessment of acid base

status; alkaline pH indicates old
sample or urinary tract infection
Specific Gravity- state of kidney
and hydration status of patient
Protein- primarily detects protein
called albumin; important indicator
in the detection of renal disease
Glucose- primarily detects glucose
(sugar); important indicator of
diabetes mellitus
Blood- red blood cells,
hemoglobin, or myoglobin (muscle
hemoglobin); sensitive early
indicator of renal disease
Ketone- normal product of fat
metabolism; increased amounts
seen in diabetes or starvation
(extreme dieting)
Bilirubin- detects bilirubin (a
product or red cell breakdown);
indicator of liver function
Urobilinogen- another by-product
of red cell breakdown; increased
amounts seen in fever, dehydration,
hemolytic anemia and liver disease
Nitrite- certain bacteria convert
normal urine nitrate to nitrite;
indicator of urinary tract infection
Leukocyte Esterase- detects
esterase enzyme present in certain

white blood cells (e.g., neutrophils,

monocytes); indicator of urinary
tract infection