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8 Cards in this Set
- Front
- Back
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LIVE/DEAD STAINS
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used for bacterial viability
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Syto 9 green fluorescent nuclei acid stain |
labels all cells in culture whether living or dead |
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propidium iodide
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-used with dead/damaged cells |
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fluorescence microscope
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1. establish: kohler, maximum resolution, optical centering, diopter correction 2. engage phase contrast for selected objective lens: - adjust focal plane as needed 3. start burner for fluorescent lamp: 4. set cube turret; WIB-wide interference blue(use for fluorescein, FITC, DiOC6, acridine orange_ WIY =wide interference yellow-green(use for rhodamine, TRITC) WU-wide interference ultraviolet (use for DAPI; Hoechst O-open 5. to control fluorescence illumination: - pull/push illumination shutter (first pull from front) to open/close it 6. set beam splitter to mid point(sends light to both oculars and camera). set camera to very low emission intensities |
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Direct Counts
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-materials: hemocytometer, coverslips, methylene blue, cultures(broth); E.coli and/or staphylococcus auereus
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counting chamber
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- chamber contains a grid that can be viewed in the microscope -the area of different squares of the grid also represent a known volume For example, each large square bounded by the double lines in the illustration is 1 square mm |
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counting the number of bacteria in 5 double lined squares
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if there are too many organisms you may have to go back and dilute your original sample further |
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