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Ruiz June 21, 2018
BS BIO BLOCK 1
1.) Types of Microscope
Brightfield A typical microscope that uses transmitted light to
microscope observe targets at high magnification.
Polarizing A microscope that uses different light transmission
microscope characteristics of materials, such as crystalline
structures, to produce an image.
Phase contrast A microscope that visualizes minute surface
microscope irregularities by using light interference. It is
commonly used to observe living cells without
staining them.
Differential This microscope, similar to the phase contrast, is used
interference to observe minute surface irregularities but at a higher
contrast resolution. However, the use of polarized light limits
microscope the variety of observable specimen containers.
Fluorescence A biological microscope that observes fluorescence
microscope emitted by samples by using special light sources
such as mercury lamps. When combined with
additional equipment, brightfield microscopes can
also perform fluorescence imaging.
Total internal A fluorescence microscope that uses an evanescent
reflection wave to only illuminate near the surface of a
fluorescence specimen. The region that is viewed is generally very
microscope thin compared to conventional microscopes.
Observation is possible in molecular units due to
reduced background light.
Laser microscope This microscope uses laser beams for clear
(Laser scanning observation of thick samples with different focal
confocal distances.
microscope)
Multiphoton The use of multiple excitation lasers reduces damage
excitation to cells and allows highresolution observation of
microscope deep areas. This type of microscope is used to
observe nerve cells and blood flow in the brain.
Structured A highresolution microscope with advanced
illumination technology to overcome limited resolution found in
microscope optical microscopes that is caused by the diffraction
of light.
2.) Parts of a Compound Microscope
Basic parts of the microscope:
Eyepiece Lens: the lens at the top that you look through. They are usually 10X or 15X power.
Tube: Connects the eyepiece to the objective lenses
Arm: Supports the tube and connects it to the base
Base: The bottom of the microscope, used for support
Illuminator: A steady light source used in place of a mirror. If your microscope has a mirror, it
is used to reflect light from an external light source up through the bottom of the stage.
Stage: The flat platform where you place your slides. 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.
Revolving Nosepiece or Turret: This is the part that holds two or more objective lenses and
can be rotated to easily change power.
Objective Lenses: Usually you will find 3 or 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. To
have good resolution at 1000X, you will need a relatively sophisticated microscope with an Abbe
condenser.
Rack Stop: This is an adjustment that determines how close the objective lens can get to the
slide. It is set at the factory and keeps students from cranking the high power objective lens
down into the slide and breaking things. You would only need to adjust this if you were using
very thin slides and you weren't able to focus on the specimen at high power.
Condenser Lens: The purpose of the condenser lens is to focus the light onto the specimen.
Condenser lenses are most useful at the highest powers (400X and above). Microscopes with in
stage condenser lenses render a sharper image than those with no lens (at 400X). I
Diaphragm or Iris: Many microscopes 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 upward 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.
3.) How to take care of a microscope
Compound Microscopes
Turn the revolving turret (2) so that the lowest power objective lens (eg. 4x) is clicked
into position.
Place the microscope slide on the stage (6) and fasten it with the stage clips.
Look at the objective lens (3) and the stage from the side and turn the focus knob (4) so
the stage moves upward. Move it up as far as it will go without letting the objective touch
the coverslip.
Look through the eyepiece (1) and move the focus knob until the image comes into focus.
Adjust the condenser (7) and light intensity for the greatest amount of light.
Move the microscope slide around until the sample is in the centre of the field of view
(what you see).
Use the focus knob (4) to place the sample into focus and readjust the condenser (7) and
light intensity for the clearest image (with low power objectives you might need to reduce
the light intensity or shut the condenser).
When you have a clear image of your sample with the lowest power objective, you can
change to the next objective lenses. You might need to readjust the sample into focus
and/or readjust the condenser and light intensity. If you cannot focus on your specimen,
repeat steps 3 through 5 with the higher power objective lens in place. Do not let the
objective lens touch the slide!
When finished, lower the stage, click the low power lens into position and remove the
slide.
NOTES:
Do not touch the glass part of the lenses with your fingers. Use only special lens paper to
clean the lenses.
Always keep your microscope covered when not in use.
Always carry a microscope with both hands. Grasp the arm with one hand and place the
other hand under the base for support.
4.) Care of the Microscope
Tip 1: Handle with care
Most microscope problems occur as a result of improper handling. When carrying your
microscope, hold it by the base and the metal support arm. Do not pick it up by the stage, as this
can cause misalignment. When transporting it, use a microscope bag.
Tip 2: Keep lenses clear of slides
When using your microscope and adjusting the focus you will need to lower the objective lens
down as far as it will go. However, you should never allow the lens to touch the slide you are
looking at. Dirty lenses can be difficult to clean.
Tip3: Clean after using immersion oil
If using immersion oil, always ensure the objectives are cleaned immediately after use.
Objective, eyepieces and condenser may be removed for cleaning. Use only lens paper and lens
cleaner. Do not use solvents.
Tip 4: Cover when not in use
All microscopes are sold with dust covers. Always keep your microscope covered when not in
use even if the microscope is stored in a cabinet. Eye tubes also need to be kept free of dust so do
not store a microscope without the eyepieces. If the microscope eyepieces must be removed,
cover the tubes with caps or a plastic bag with a rubber band around the eye tube.
Tip 5: Look after the bulb
After using the microscope, turn off the illuminator and wait for it to cool for several minutes
before putting it away. By allowing the bulb to cool you will extend its life. When turning the
microscope on and off, use the switch not the power point. Do not switch the microscope on
while using full light intensity. Never touch the bulb with your fingers as the body oils can burn
into the bulb and reduce its life. Use a tissue. Keep a store of replacement bulbs and always use
the correct bulb.
Tip 6: Store in a clean, dry place
Make sure you do not store your microscope in an area that has corrosive chemical fumes that
can destroy lenses or metal parts or beside solutions that may leak. Salt air and pervasive damp
can also cause damage over time. Make sure your cabinet is ventilated.
Tip 7: Only use special lens paper or wipes for cleaning the lenses
Microscope lenses can easily be scratched and should be treated with great care. Use an aspirator
to remove dust. Sticky residue can be removed with lens paper moistened with distilled water or
lens cleaning solution and rubbed gently using a circular motion. Never use sharp instruments or
anything abrasive on the microscope lenses.
Tip 8: Keep your User's Manual and wrenches in a safe place
Each microscope should come with a user's manual and specialist wrenches as required. Always
consult the User's Manual before making any adjustments to your microscope and use the
wrenches provided. Never overtighten or use force when performing any maintenance on your
microscope, or use inappropriate tools. This can damage the parts.
Tip 9: Perform an annual maintenance check
On an annual basis moving parts on the microscope should be cleaned and lubricated. Clean
grease and dirt from sliding surfaces using a clean cloth. Apply a very thin layer of lithiumbased
grease to the sliding surfaces. Do not grease the teeth of the rack and pinion gears. Inspect the
power cords and plugs for safety and stock up on a supply of replacement bulbs.
Tip 10: Have your microscope serviced professionally
A rule of thumb for frequency of servicing is every 200 hours of use or every 3 years, whichever
comes first.
Sources:
1.) https://www.microscopeshop.com.au/microscope_maintenance.html
2.) https://www2.mrclmb.cam.ac.uk/microscopes4schools/microscopes2.php
3.) http://enfo.agt.bme.hu/drupal/en/node/8862
4.) https://www.keyence.com/ss/products/microscope/bzx700/study/principle/002/index.jsp