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chapter 2 - Chapter 2 Microscopy Types of Microscopy Light...

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Chapter 2: Microscopy
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Types of Microscopy Light Microscopes: Use light rays to help visualize the specimen, up to 1000X TM Electron Microscopes: Uses a beam of electron radiation and capture to produce images Best resolution and greater magnification than light microscopes Requires expensive chemical preparation of specimens and sophisticated equipment
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Light Microscopy 1. Brightfield: Images appear dark, background is white Specimens need either natural pigments or to be stained; not good for live material Light passes directly through and around the specimen
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Light Microscopy 2. Darkfield: Images appear bright, background is dark No need for stain or pigments; excellent for live material Light refracted by the specimen is collected and passed into the objective lens
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Light Microscopy 3. Phase Contrast: Images appear bright, background is dark No need for stain or pigments; excellent for live material Measures variations in light intensity via density of specimen **more common than darkfield
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Light Microscopy 4. Fluorescence: Uses dyes or natural pigments that produce light as a means to visualize the specimen Depending on fluorchrome, can be used with live material Uses one wavelength of light to excite the dyes; special filters are used to exclude certain wavelengths of emitted light so that a small region of wavelengths can be viewed (i.e. blues) ***especially helpful for organisms that naturally bioluminesce
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Electron Microscopy 1. Transmission: Electron beam penetrates the specimen and then scatters Electrons bounce back to the detector and are collected to produce images Intracellular detail often studied in this manner
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Electron Microscopy 2. Scanning: Electron beams hit surface of metal-coated specimen and bounce back to detector to produce image
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Microscopy Terms Refraction: bending of light rays as they pass from one type of substance (i.e. air) to another (i.e. glass) Refractive index: a measure of the degree rays are bent Lens:
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