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reason to learn about microbiota
#1
to gain an understanding of different bacteria at specific body sites. which provide greater insight to possible infections that result from injury
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reason to learn about microbiota #2
to gain knowledge of the possible source and significance of bacteria isolated from clinical infections.
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reason to learn about microbiota #3
to gain knowledge of consequences of overgrowth of those bacteria normally absent at a specific body site.
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reason to learn about microbiota #4
to be aware of bacteria's play in stimulating the host immune system
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what constitutes a possitive reaction with 2O2?
bubbles of oxygen gas is a posstive reaction
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gram stain purpose of each reagent #1
primary stain cyrstal violet interacts with the peptidglycan in the cell wall
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gram stain purpose of each reagent #2
the mordant iodine increases the interaction of the cyrstal violet and the cell wall
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gram stain purpose of each reagent #3
the decolorizer remives the crystal violet from the gram negative bacteria cell wall, but not the gram possitive.
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gram stain purpose of each reagent #4
the secondary stain safrain counterstains the gram negative bacteria with a red or pink color
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gram stain procedure
step#1
- transfer a loop of stock culture to a slide.
- air dry.
- heat fix and place on a rack.
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gram stain procedure step#2
- flood slide with crystal violet for one minute.
- rinse slide with water and place back on rack.
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gram stain procedure step#3
- flood slide with iodine for one minute.
- rinse with water.
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gram stain procedure step#4
- drip decolorizer onto the slide and let it run off for 5 seconds
- rinse with water and place back on rack
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gram stain procedure step#5
- flood slide with saffranin for one minute.
- rinse with water and blot dry.
- observe under the microscope
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kinyoun acid-fast staining proceedure step #1
- transfer stock culture to slide.
- air dry. heat fix.
- place slide on rack and flood with carbofuschsin for 5 min.
- rinse with water.
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kinyoun acid-fast staining proceedure step#2
- drip decolorizer on slide to rinse the stain from the specimen, until there is just a tint of color running off.
- rinse with water.
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kinyoun acid-fast staining proceedure step #3
- flood with brillant green stain for 2 min.
- rinse with water, blot dry.
- observe under microscope.
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kinyoun acid-fast staining reagents step #1
primary stain, carbon fusion, is retained by the acid fast waxy complex in the cell wall
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kinyoun acid-fast staining reagents step #2
the decolorizer removes the carbonfuschsin from the non-acid fast bacteria.
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kinyoun acid-fast staining reagents step #3
the counter stain, brilliant green, stains the non-acid fast bacteria a greenish- blue
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2O2 superoxide equation
- 2O-2+2H===>O2+H2O2
- superoxide/dismutase= oxygen + hydrogen peroxide
- 2H2O2===> 2H2O+O2
- catalaseor/peroxidase= water + free oxygen
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"OF" proceedure materials
- 2 "OF" tubes,
- needles,
- stock cultures,
- mineral oil.
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How to perform an "OF" proceedure.
- using needles innoculate both tubes of "OF" with stock culture by stabbing each tube 3-4 times, but not all the way to the bottom.
- Overlay one tube with mineral oil, leave both caps loose.
- incubate overnight at 35 degrees
- leave them all open.
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