Cytoskeleton Methods and Protocols by Torsten Wöllert, George M. Langford (auth.), Ray H. Gavin

By Torsten Wöllert, George M. Langford (auth.), Ray H. Gavin (eds.)

In the 10 years because the book of the 1st version, nice advances in fluorescent labeling, optics, and pattern guidance have considerably more suitable the imaging strength of microscopy, taking into account a continuous refinement of our figuring out of the cytoskeleton as a dynamic synergy of elements. In Cytoskeleton tools and Protocols, moment Edition, across the world well known specialists current options which mirror a number of the contemporary technological advances in experimental instruments for cytoskeleton learn with emphasis on animal, plant, protist, and fungal version platforms. This state-of-the-art quantity includes equipment for live-cell imaging, fluorescence microscopy, electron microscopy, research of phone and organelle motility, isolation of cytoskeleton parts, and proteomics, among different themes. As a quantity within the hugely winning Methods in Molecular Biology™ sequence, chapters comprise introductions to their respective topics, lists of the required fabrics and reagents, step by step, easily reproducible laboratory protocols, and notes that offer unpublished technical info on troubleshooting and warding off identified pitfalls.

Up-to-date and accomplished, Cytoskeleton tools and Protocols, moment Edition serves as a terrific advisor to scientists who desire to proceed this fruitful and critical organic study.

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Swayne, Thomas G. Lipkin, and Liza A. Pon Summary This chapter describes labeling methods and optical approaches for live-cell imaging of the cytoskeleton and of a specific organelle–cytoskeleton interaction in budding yeast. Key words: Yeast, Mitochondria, Fluorescent proteins, Immunofluorescence, Vital staining, Microscopy, Live-cell imaging, Deconvolution 1. Introduction The actin and microtubule cytoskeletons of budding yeast, and their cargos, were first detected by staining fixed cells with dyes or antibodies.

G. (1998). Relationship between microtubule dynamics and lamellipodium formation revealed by direct imaging of microtubules in cells treated with nocodazole or taxol. Cell Motil Cytoskeleton 41, 325–340 12. J. (2006). Reassembly of contractile actin cortex in cell blebs. J Cell Biol 175, 477–490 13. B. K. (1991). Xenopus laevis: practical uses in cell and molecular biology. Methods Cell Biol 36, 679–681 14. M. (2007). Versatile fluorescent probes for actin filaments based on the actin-binding domain of utrophin.

5 h after fertilization. 3. Microinjection of Xenopus laevis Embryos We microinject Xenopus embryos at the 1-, 2-, 4- or 8-cell stage using a PLI-100 picoinjector (Harvard Apparatus; see Note 5). 1). , morpholino) see Note 6. When injecting RNA, we use measures to limit RNases by cleaning injection areas with RNaseZAP® (Ambion), using RNase-free reagents and by wearing gloves at all times. 1. 5 h after fertilization (at 17°C). 1. Pull needles using 10 µL glass capillaries and a micropipette puller.

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