By Peter J. Verveer
This quantity presents an outline of complicated fluorescence microscopy, masking a wide variety of tools. each one bankruptcy makes a speciality of a distinct procedure and offers a realistic consultant for program in organic structures. Written within the hugely winning Methods in Molecular Biology sequence structure, chapters comprise introductions to their respective themes, lists of the required fabrics and reagents, step by step, comfortably reproducible laboratory protocols, and tips about troubleshooting and averting identified pitfalls.
Authoritative and state-of-the-art, Advanced Fluorescence Microscopy: tools and Protocols seeks to supply scientists with equipment for organic structures which are of curiosity.
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Additional resources for Advanced Fluorescence Microscopy: Methods and Protocols
12). pii: 678. 3791/678. Schaffer CB, Friedman B, Nishimura N et al (2006) Two-photon imaging of cortical surface microvessels reveals a robust redistribution in blood flow after vascular occlusion. PLoS Biol 4:e22 21. Kleinfeld D, Mitra PP, Helmchen F et al (1998) Fluctuations and stimulus-induced changes in blood flow observed in individual capillaries in layers 2 through 4 of rat neocortex. Klunk WE, Bacskai BJ, Mathis CA et al (2002) Imaging Abeta plaques in living transgenic mice with multiphoton microscopy and methoxyX04, a systemically administered Congo red derivative.
Neubauer cell counting chamber. 35 mm glass-bottomed imaging dishes (MatTek Corporation, USA). 49, Nikon). l) set at 37 °C for live-cell imaging. 3. Green diode 561 nm laser. 4. 12-bit iXon 1 M EM-CCD camera. 5. 2 software. Trypsinise a T75 flask of MDA-MB-231 cells and count the number of cells per 1 ml using a counting chamber. Seed 6 × 105 cells onto each 35 mm glass bottom dish and incubate in complete medium at 37 °C, 5 % CO2 for 24 h. On day 2, transiently transfect cells with paxillin-RFP using Lipofectamine 2000 according to the manufacturer’s instructions using a ratio of 4 μg DNA to 10 μl Lipofectamine 2000 per 35 mm dish of cells.
Mattheyses AL, Simon SM, Rappoport JZ (2010) Imaging with total internal reflection fluorescence microscopy for the cell biologist. Axelrod D (2001) Total internal reflection fluorescence microscopy in cell biology. Axelrod D (2008) Total internal reflection fluorescence microscopy. Rappoport JZ, Simon SM (2003) Real-time analysis of clathrin-mediated endocytosis during cell migration. J Cell Sci 116:847–855 5. Rappoport JZ, Benmerah A, Simon SM (2005) Analysis of the AP-2 adaptor complex and cargo during clathrin-mediated endocytosis.
Advanced Fluorescence Microscopy: Methods and Protocols by Peter J. Verveer