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Intro to Cell Biology

20 microscopy visualization of small objects using

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Microscopy Visualization of small objects using light or electrons to illuminate and excite, lenses to magnify, and detectors to sense Magnification- The number of times an image’s sized is enlarged Resolution- ability to distinguish two closely spaced points 21
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Bright Field Light Microscopy 22
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Antibodies Heavy Chain- Invariable region Light Chain- Variable region macromolecule of interest= antigen Isolate Antibodies from animal and purify those specific to antigen Polyclonal- mix of antibodies that recognize different epitopes on an antigen Monoclonal- antibody that recognizes a single epitope on an antigen Primary antibody- Recognizes macromolecule of interest Secondary antibody- Recognizes heavy chain of the primary antibody 23
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Fluorochromes, Fluorescent Dyes, and Proteins Link fluorescent proteins to protein of interest = one continuous fusion protein Protein of interest gfp 24
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Fluorescent Microscopy 25
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Confocal Microscopy and Deconvolution 26
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FACS: Fluorescence-Activated Cell Sorting 27
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Cell Culture Cell culture conditions mimic in vivo conditions -food/energy source -signal to grow (growth factors) -extracellular substrate 28
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Western Blots Large proteins Small proteins 30
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Immunoprecipitation Examine interaction of two macromolecules Precipitate Protein X Western Blot for Protein Y 31
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20 Microscopy Visualization of small objects using light or...

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