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PREPARING SAMPLE, Microscope: FOCUSING, OCULAR MICROMETER Calibration,…
PREPARING SAMPLE
Bacteria
Yeast 0.5% activated
baker's yeast cell suspension
Protist/single-celled eukaryote: pond water, hay infusion/prepared slide
Hydrilla leaf cell
Human cheek cell
Scrape w/ toothpick
Wipe end on glass surface
drop of water
leaf on slide
drop of water on it
Pasteur pipette, a drop on slide
Place end of coverlsip on slide
Lower other end on top of sample
Stain (YEAST & CHEEK ONLY)
drop methylene blue next to coverslip
draw it under coverslip by placing clean tissue on opposite side
Microscope: FOCUSING
Observe samples under microscope
Clean mirrors n lenses w/ lens paper
Objective must be far away from stage as possible
Place slide on stage
center object on focus over hole
Secure w/ stage clips
**If microscope has illuminator, turn on
Diaphragm should be wide OPEN as possible
Focus object into view
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**If microscope has mirror, adjust so field is evenly illuminated
OCULAR MICROMETER Calibration
Remove 10x ocular from microscope
Unscrew eye lens
Place micrometer disc w/in ocular
Screw back eye lens, insert back to microscope
Place stage micrometer on stage
Focus on some portion of scale
examine stage micrometer to distinguish 0.1 mm and 0.01 divisions
Adjust field so 0 line of ocular and stage micrometer are superimposed
DONT MOVE STAGE MICROMETER | find another pt at extreme right/down were other lines are superimposed
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MAGNIFICATION
Mount slide w/ samples
Draw entire specimen to length of 3cm
Indicate magnification of your illustrations
CELL COUNT: HEMOCYTOMETER
1.5mL tube
200 uL yeast suspension
20 or 10 uL methylene blue stain
Mix 2gether
Incubate ~1 min at room temp
**Count cells within 10 mins, else cell death
Wet supports (rails) with water
Place coverslip on hemocytometer
Using, Pasteur pipette, + drop of methylene blue/cell mixture to a hemocytometer
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