Molekulární biologie (KBC/MBIOG) Ivo Frébort Alberts et al. (2008) Molecular Biology of the Cell, 5th ed. Garland Science, New York 12. Methods of molecular biology II: Visualizing cells
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Molekulární biologie (KBC/MBIOG) Ivo Frébort Alberts et al. (2008) Molecular Biology of the Cell, 5th ed. Garland Science, New York 12. Methods of molecular.
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Molekulární biologie(KBC/MBIOG)
Ivo Frébort
Alberts et al. (2008) Molecular Biology of the Cell, 5th ed.Garland Science, New York
12. Methods of molecular biology II:Visualizing cells
A sense of scale
A light microscope
Interference and edge effects
Numerical aperture
Obtaining contrast
Four types of light microscopy
Bright-field microscopy
Phase-contrast microscopy
Differential-interference-contrast microscopy
Dark-field microscopy
Image processing
Tissue sectioning
Fluorescence microscopy
Fluorescence dyes
Immunofluorescence
Image deconvulsion – removing the blur by computing
The confocal fluorescence microscope
Conventional vs. confocal fluorescence microscope
Confocal microscopy allows 3D recontruction of objects
Trichomes of Arabidopsis containing talin-GFP
Green fluorescent protein can be used to tag individual proteins in living cells and organisms
GFP from jellyfish Aqueoria victoria
Fluorescence resonance energy transfer (FRET)
Visualizing cell dynamics using caged molecules
Determining microtubule flux in the mitotic spindle with caged fluorescein linked to tubulin
Dynamic changes and photoactivation of GFP fluorescence
Fluorescence recovery photobleaching (FRAP)
Visualizing living cells: light-emitting indicators
Sperm entry into a fish egg visualized with aequorin/Ca2+
Neurone cell from the brain of a guinea pig – indicator fura-2
Visualizing Ca2+ concentration by a fluorescent indicator
Introducing large molecules into cells
Laser tweezers
manipulating objects with higher refractive index within the cell
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Total internal reflection fluorescence (TIRF) microscopy can visualize single molecules
Single molecules can be manipulated by atomic force microscopy (AFM)