Journal article
Fluorescence microscopy of actin- and microtubule-associated septins in mammalian cells
Methods in cell biology, v 136
2016
PMID: 27473913
Abstract
Septins are a major component of the mammalian cytoskeleton. Septins associate with filamentous actin (F-actin) and microtubules, but the nature and significance of these interactions are not well understood. Fluorescence microscopy of F-actin- and microtubule-associated septins in fixed and living cells has been instrumental in uncovering septin functions in cellular morphogenesis and cytoskeleton-dependent processes (eg, cell division, cell migration). Here, we provide a detailed methodology for the visualization of endogenous septins by immunofluorescence microscopy, discussing sample preparation and reagents that are critical for optimal staining. In addition, we review approaches for the construction and expression of fluorescent septins and their time-lapse imaging with F-actin and microtubules. The recommended methodology is adaptable for high- and superresolution imaging of mammalian cells with various instrumentation, including wide-field and confocal microscopy as well as total internal reflection fluorescence and structured illumination microscopy.
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Details
- Title
- Fluorescence microscopy of actin- and microtubule-associated septins in mammalian cells
- Creators
- E T Spiliotis - Drexel University, Philadelphia, PA, United StatesE P Karasmanis - Drexel University, Philadelphia, PA, United StatesL Dolat - Drexel University, Philadelphia, PA, United States
- Publication Details
- Methods in cell biology, v 136
- Publisher
- Elsevier; United States
- Grant note
- F31 CA176910 / NCI NIH HHS R01 GM097664 / NIGMS NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biology
- Web of Science ID
- WOS:000383711700015
- Scopus ID
- 2-s2.0-84964334064
- Other Identifier
- 991014877755304721
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemistry & Molecular Biology
- Cell Biology