Polarized Fluorescence Microscopy to Study Cytoskeleton Assembly and Organization in Live Cells.
The measurement of not only the location but also the organization of molecules in live cells is crucial to understanding diverse biological processes. Polarized light microscopy provides a nondestructive means to evaluate order within subcellular domains. When combined with fluorescence microscopy and GFP-tagged proteins, the approach can reveal organization within specific populations of molecules. This unit describes a protocol for measuring the architectural dynamics of cytoskeletal components using polarized fluorescence microscopy and OpenPolScope open-access software (http://www.openpolscope.org). The protocol describes installation of linear polarizers or a liquid crystal (LC) universal compensator, calibration of the system, polarized fluorescence imaging, and analysis. The use of OpenPolScope software and hardware allows for reliable, user-friendly image acquisition to measure and analyze polarized fluorescence.
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- Software
- Septins
- Saccharomyces cerevisiae Proteins
- Saccharomyces cerevisiae
- Microbial Viability
- Image Processing, Computer-Assisted
- Green Fluorescent Proteins
- Fluorescence Polarization
- Cytoskeleton
- 3101 Biochemistry and cell biology
Citation
Published In
DOI
EISSN
Publication Date
Volume
Start / End Page
Location
Related Subject Headings
- Software
- Septins
- Saccharomyces cerevisiae Proteins
- Saccharomyces cerevisiae
- Microbial Viability
- Image Processing, Computer-Assisted
- Green Fluorescent Proteins
- Fluorescence Polarization
- Cytoskeleton
- 3101 Biochemistry and cell biology