By Diana P. Bratu (auth.), Douglas J. Taatjes, Brooke T. Mossman (eds.)
A various number of cutting-edge equipment for the microscopic imaging of cells and molecules. The authors conceal a large spectrum of complimentary strategies, together with such equipment as fluorescence microscopy, electron microscopy, atomic strength microscopy, and laser scanning cytometry. extra simply reproducible protocols on confocal scanning laser microscopy, quantitative computer-assisted photograph research, laser-capture microdissection, microarray snapshot scanning, near-field scanning optical microscopy, and mirrored image distinction microscopy around out this eclectic selection of state of the art imaging thoughts now on hand. The authors additionally speak about preparative tools for debris and cells by way of transmission electron microscopy.
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Extra resources for Cell Imaging Techniques: Methods and Protocols
Specific labeling of the cytoplasm with a fluorescent Ca2+ dye can be achieved by microinjection of individual cells with a membrane-impermeable form of the dye, reversibly permeabilizing the plasma membrane of a population of cells in the presence of the potassium salt form of the dye using scrape loading or osmotic techniques (see ref. 7 for a review). Microinjection is labor intensive, technically demanding, and does not allow the study of a large number of cells simultaneously. The latter approaches risk damaging cells and do not enable loading of subcellular compartments such as organelles with Ca2+ indicators.
Proc. SPIE 4620, 148–156. 17. Blab, G. , Lommerse, P. H. , Harms, G. , and Schmidt, T. (2001) Two-photon excitation action cross-sections of the autofluorescent proteins. Chem. Phys. Lett. 350, 71–77. 18. , Gengfeng Zheng, and Sunney Xie, X. (2002) Laserscanning coherent anti-Stokes Raman scattering microscopy and applications to cell biology. Biophys. J. 83, 502–509. 19. , Sheppard, C. J. , and Xu, P. (2002) Resolution of second harmonic generation microscopy. Durban, South Africa. Proc. 15th Int.
1. Schematic arrangement of lens and detector arrangements on our Leica TCS2 MP/DMIRB inverted-microscope system. The two dual-channel detectors are identical and have removable filter cubes, so that different combinations are easy to assemble. The 650DCLP dichroic is mounted in a cube of the wide-field epifluorescence filter changer and directs all fluorescence returning through the objective to the detector. When this is rotated out of position, the normal confocal detection system can be used.