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Lecture2-2 - 1 How to study cells Tools used to answer...

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1 How to study cells Tools used to answer biological questions. The methods are important, but it's what we discover with them that's really interesting. 1. Microscopy (chap 1) Role of microscopy in Cell Biology - Invention of light microscopes in 1600s - cells visible for the first time - sole tool to study cells for centuries - light microscopy is still a valuable tool to cell biologists What microscopes do: - designed for magnification, resolution, & contrast - wavelength of light limits resolution - sizes of cells and cell components Types of microscopy: 1. light microscopy a. conventional wide field b. phase contrast, differential interference contrast c. darkfield - staining - Preparing cells/tissues for light microscopy: fixation sectioning d. fluorescence microscopy fluorescent dyes (see figure) can be used to identify molecular structures. - Viewing fluorescent cells: (1) Epi-fluorescence illumination (see figure) (2) Laser-scanning confocal microscopy (see figure) - Immunofluorescence - use of antibodies - Indirect immunofluorescence - Fluorescent proteins
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2 fluorescein epifluorescence confocal excitation emission
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