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38 Cards in this Set
- Front
- Back
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condenser
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concentrates beam of light
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iris diagram
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regulates amount of light
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fine & coarse focus
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moves stage up or down to change focal distance
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resolution
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ability to see details of object
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depth of field
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vertical distance through which an object is in focus
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working distance
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distance between slide & objective lens
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size of field
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diameter of field of view
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illumination
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light source, best a 4x (most light in)
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Function of Immersion Oil
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Has same refractive indez as the glass lens & glass slide. This prevents distortion because light waves follow a homogeneous path.
Basically, immersion oil prevents refraction of light. |
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What are the 5 anaerobic methods?
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Gaspak
Candle Jar Thioglycollate broth Slant Deep spike into center- |
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Gaspak
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gets rif of oxygen, uses asorbic acid pack (antioxidant)
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Candle Jar
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not an anaerobic environment,
microaerophiles grow, Carbon Dioxide levels High |
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Thioglycollate broth
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no oxygen below reasurin ring
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Slant
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streak outside and spike into center- anaerobic environment on bottom
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Deep Spike to center
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anaerobic environment on bottom
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How do you prepare a streak plate
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To streak a plate for isolation, start by labeling the plate with details about contents & date,
divide plate into 4 quadrants with crayon. Pick up test tube with non-dominant hand. Remove lid of test tube with non hand holding the loop. With the little finger of this hand, remove the lid from the test tube. Be careful not to touch the inside area of the lid. Pass the test tube through the flame a few times. Flame your loop and get a loopful of the bacteria. Carefully replace the lid and put the test tube in the test tube coffin. Crack open the petri dish and Streak the culture in the first quadrant three or four times. Flame loop, then go over first section again and spread bacteria along the second section. Then flame loop again. This time spread the loop a few times in the second quadrant then into the third, Repeat with third section and fourth. Final quadrant should have an isolated colony. |
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Why are smears heat fixed?
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Because the heat coagulates the organisms' proteins causing the bacteria to stick to the slide
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Simple Stain
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One color
positive stain-- color attaches to cell wall (crystal violet, methylene blue, safranin) negative stain- cell wall unstained, background colored (congo red, nigrosin) Simple Stains react with all microbes in an identical fashion. They increase contrast so that morphology, size, and arrangement of organisms can be determined |
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Differential Stain
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2 or more dyes, shows different color for different categories of organisms
ex- acid fast, g+ & g- Give varying results depending on organism being treated. These results are often helpful in identifying microbe |
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Liquid Smear
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label slide
flame loop- suspend tube take off lid- pass lid through flame 2 to 3x obtain sample using loop reflame tip of tube, replace tube with lid in coffin spread monolayer on slide flame loop air dry slide heat fix |
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solid smear
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label slide
flame loop place small drop of deionized H20 on loop and transfer D20 to slide Flame loop and cool obtain small "bite" of colony on loop mix with DH20 to make monolayer (get the clumps out) Air Dry Heat fix by gently passing slide though flame |
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Reagents of gram Stain
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Crystal violet (primary stain)- imparts color to all cells
Gram's Iodine- a mordant, substance added to staining solution to make it stain more intensely Acetone alcohol (decolorizing agent)- removes purple from some cells Gram's Safranin- (counterstain- contrasts with color or primary)- a basic red dye, gram negative cells are colorless until counterstained, which turns them pink |
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Acid Fast Organisms
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Red
retain red color because carbolfuchsin is more soluble in cell wall lipids than in acid-alcohol |
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Non Acid Fast Organisms
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Blue
non acid fast bacteria whose cell walls lack lipid components, the carbolfuchsin is rapidly removed during decolorization, leaving cells colorless. After methylene blue counterstain, they appear blue |
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Reagents of acid fast
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red dye carbolfuschin
DH20 Acid Alcohol Loeffler's Methylene Blue |
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Purpose of using heat during acid fast staining process
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enhances penetration and retention of die. Heat melts away the waxy lipid cell wall so dye can penetrate
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Common genera of endospore producing bacteria
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Clostridium
Bacillus |
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Colors of spores and cells in methods (schaeffer-fulton and gram stain) used
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Schaffer- malachite green spores and pink vegetative cells when viewed on oil
Spores- Gram neg. (colorless) |
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2 advantages of using negative staining instead of positive
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Only way to see delicate organisms like spirochete
Doesn't distort size and shape Fast and simple |
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What's the difference between negative & positive staining?
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Negative- you dye the background and organisms will be clear, has a negative charge and is repelled from the cell wall
Positive- background not colored, organism is dyed |
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Genus name of fungal organism when seen under a microscope
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rhizopus, penicillium
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What medium is grown in our lab to grow fungi
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Sabourand's dextrose
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Why is Sabourand's dextrose selective for fungi
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agar is low in pH and high in sugar. Fungi can grow on it but don't really love it. Bacteria can't grow on it so it's not overwhelmed by bacteria.
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Broken glass container
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under hood, back of room, cardboard box
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plate coffins
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Red bags
dump plates here in back under hood |
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used slide container
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glass container with liquid at station
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disinfectant beakers for toothpicks
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at your station under hood
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micropipette tips
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autoclave or biohazard bags (sharpe)
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