Document Detail

Fluorescence fluctuation approaches to the study of adhesion and signaling.
MedLine Citation:
PMID:  23280111     Owner:  NLM     Status:  MEDLINE    
Cell-matrix adhesions are large, multimolecular complexes through which cells sense and respond to their environment. They also mediate migration by serving as traction points and signaling centers and allow the cell to modify the surroucnding tissue. Due to their fundamental role in cell behavior, adhesions are germane to nearly all major human health pathologies. However, adhesions are extremely complex and dynamic structures that include over 100 known interacting proteins and operate over multiple space (nm-μm) and time (ms-min) regimes. Fluorescence fluctuation techniques are well suited for studying adhesions. These methods are sensitive over a large spatiotemporal range and provide a wealth of information including molecular transport dynamics, interactions, and stoichiometry from a single time series. Earlier chapters in this volume have provided the theoretical background, instrumentation, and analysis algorithms for these techniques. In this chapter, we discuss their implementation in living cells to study adhesions in migrating cells. Although each technique and application has its own unique instrumentation and analysis requirements, we provide general guidelines for sample preparation, selection of imaging instrumentation, and optimization of data acquisition and analysis parameters. Finally, we review several recent studies that implement these techniques in the study of adhesions.
Alexia I Bachir; Kristopher E Kubow; Alan R Horwitz
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in enzymology     Volume:  519     ISSN:  1557-7988     ISO Abbreviation:  Meth. Enzymol.     Publication Date:  2013  
Date Detail:
Created Date:  2013-01-02     Completed Date:  2013-06-05     Revised Date:  2013-07-11    
Medline Journal Info:
Nlm Unique ID:  0212271     Medline TA:  Methods Enzymol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  167-201     Citation Subset:  IM    
Copyright Information:
Copyright © 2013 Elsevier Inc. All rights reserved.
Department of Cell Biology, University of Virginia, Charlottesville, Virginia, USA.
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MeSH Terms
Fluorescent Dyes / chemistry
Proteins / chemistry*
Signal Transduction*
Spectrometry, Fluorescence / methods*
Grant Support
Reg. No./Substance:
0/Fluorescent Dyes; 0/Proteins

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