Exercise #1: The Microscope Fine adjustment knob High power objective

- to view structures at diff. - to view finer details of

Smaller than 1/10 mm - cannot be seen by naked eye depths of field specimen
- for final focus of specimen
1. Light microscopy - uses light to illuminate specimens Mirror (plane - natural; Scanning objective
(dark specimen, bright background) concave - artificial)
2. Compound light microscope - use of 2-lens system Condenser Oil immersion objective
(thin, minute specimens) - focuses light to specimen
3. Dark-field - collects light rays to single
4. Phase contrast cone of light
5. Electron microscopy
III. Eyepiece micrometer
I. Microscope handling and care
Eyepiece micrometer
a) One hand - arm; one hand - base - used to measure quantities such as length, diameter, distance
Upright position - has graduated scale with NO INHERENT VALUE
3 cm from the edge
Stage micrometer
b) Prevent liquid contact w/ any part of the microscope - slide provided with a scale of KNOWN VALUES
Always use a cover slip when examining specimens
Remove excess fluids when preparing wet mounts
Calibration constant Size (l) of objects
c) Blow air using aspirator or lens cleaner bulb to remove
dirt from lenses SMDx0.01mm m l = EMDobject x c
Use d. H2O, 90-100% ethanol, or acetone in wiping off c( )(1000 )
EMD mm
residue
Use Xylene to remove thick, gummy substances

d) Before returning; mirror - vertical position facing IV. Magnification and resolution
sideways
Set to lowest possible objective (LPO; 4x) - 1 cm above Magnification
stage - process of producing enlarged images of specimens
- ratio of the size of a drawing to that of the object drawn
II. Compound Light Microscope (value written beneath the drawing, "x", computed value)

e) Parts of the microscope Resolution

Base
- U shaped
- support structure
- used in carrying
Pillar
- vertical extension
Arm
- where upper parts rest
- supports the microscope
- used for carrying
Inclination joint
- connects arm to pillar
Stage
- base of arm
Stage clips
- located at the base
Coarse adjustment knob
- to raise/lower body tube
- to initially focus specimen
- ability of microscope to distinguish 2 points of an image
Iris diaphragm
- below the condenser Linear magnification (total magnification)
- provides contrast - refers to relative size of FINAL IMAGE (after light passes
through)
Substage - product of magnifying powers of each lens system
- condenser + iris diaphragm
Substage knob  Magnifying power of ocular and objectives - "x"
- to raise/lower substage  Numerical aperture (NA) - between 0 and 1; denotes
resolving power of an objective; helps determine
resolution of objective
Ocular/Eyepiece
- inserted to draw tube
Body tube
- below draw tube
Revolving nosepiece
- holds objectives
Low power objective
- shorter (4x)
Take-home questions
1. What is a stereomicroscope?
2. What is the length of the body tube? What is its significance
in the function of the microscope? What happens to the path of
light as it traverses the body tube?
3. How is the working distance of an objective lens related to
its use? What is the numerical aperture, and how is it related to
the resolution of an image?
4. How often do we need to calibrate the eyepiece
micrometer? Why? What is the average calibration constant
assigned to a combination of 10x ocular and LPO? What about
a 10x ocular and HPO?