Lecture Notes 11-5-08 - Dark Field Microscopy Results in...

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Dark Field Microscopy Results in very high contrast Useful for visualizing cells that have very low intrinsic contrast Such as cells in suspension Mitochondria Nucleus Stains are essentially invisible with this approach
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The stained portions of the cell reduce the amplitude of light waves passing through them (A) making the light less bright Light passing through the unstained living cell experiences a phase change (with very little change in amplitude), which causes interference (B). Small phase differences can be made visible by exploiting interference effects using a phase-contrast or a differential-interference-contrast microscope
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Staining Cells In order to preserve cells, they must be rapidly killed in a way that immobilizes them Fixatives cross-link adjacent proteins together The most effective method of tissue fixation is via vascular perfusion Most tissue samples are too thick to be examined directly They must first be cut into very thin slices, or sections,
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Lecture Notes 11-5-08 - Dark Field Microscopy Results in...

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