PLPA 3004 Lecture Notes - Lecture 1: Ernst Abbe, Eyepiece, Oil Immersion

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Published on 6 Oct 2016
PLPA 3004 – BIOL 3004 Lab #1
1660’s Antony von Leeuwenhoek and Robert Hooke
Ernst Abbe construction of microscope on a scientific basis – first to realize the restrictions on the
microscope imposed by the nature of optical systems.
Nomarski microscopes utilize wave interference properties of light to enhand the image of living
material and structures without the necessity of staining
Parts of the Compound Microscope
Ocular lens, turrent (grab) objective lens, stage, condenser (focus resolution), Light Iris diaphragm ring
(more/less light), base, stage motion knobs, course focus, fine focus, light switch and knob
Theory of microscope operation
Compound microscope optical system in which a double magnification takes place.
Smallest resolvable object diameter = wavelength of light
2 x numerical aperature
With green light: Smallest resolvable object that can be seen: 0.2micrometers or 0.00022 cm in diameter
To increase resolution, wavelength or the numberical aperture must be manipulated
Oil immersion an objective lens of high magn. Requires use of immersion oil.
1.Hold arm at base 2. Place at least 4 in from edge 3. Plug in and turn on light 4. Put on slide on stage at
lowest power first (10X) 5. Adjust condenser 6. Remove eyepiece, replace eyepiece 7. Rotate turret,
clicks into place 8. Use oil immersion objective (100X) 9. Clean with dry lens paper 10. Clean, dust, put
on low power, cover and store
Stereo-microscope – acts as 2 compound micros. Working side by side. Principle of parallax (viewing an
object from 2 different angles) view is 3 dimensional. Care is similar
Scrape mount used for detecting pathogen structures on a leaf surface with drop of water. Dry material
or hydrophobid, add alcohol to allow water to penetrate. Scrape leaf surface, fluid creates suplension,
drop to slide, glass slide, cover slip.
Tape mount useful for same purposes as a scrape mount but use double-stick tape, place slide, press
leaf surface, drop of water, cover slip
Suash mount used for observing internal contents of opaque microscopic objects with drop of fluid,
transfer needle, tease out material of plant tissue, cover slip, back of pencil, squash object with eraser,
cover slip
Freehand section used for observing the relationshjp of pathogen structures to plant tissue. Use 2 slide
– place tissue on slide with labeling tape, use second micros. Slide so one end serves as straight edge
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