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Fluorescence Fluctuation Approaches to the Study of Adhesion and Signaling

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,... Full description

Journal Title: Methods in Enzymology 2013, Vol.519, p.167-201
Main Author: Bachir, Alexia I
Other Authors: Kubow, Kristopher E , Horwitz, Alan R
Format: Electronic Article Electronic Article
Language: English
Subjects:
Quelle: Gale eBooks
Publisher: United States: Elsevier Science & Technology
ID: ISSN: 0076-6879
Link: https://www.ncbi.nlm.nih.gov/pubmed/23280111
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recordid: cdi_gale_vrl_7001800015
title: Fluorescence Fluctuation Approaches to the Study of Adhesion and Signaling
format: Article
creator:
  • Bachir, Alexia I
  • Kubow, Kristopher E
  • Horwitz, Alan R
subjects:
  • Aggregation state
  • Article
  • Cell adhesion
  • Cell migration
  • Cells (Biology)
  • CLSM
  • Complex stoichiometry
  • Fluorescence
  • Fluorescence fluctuation
  • Fluorescence imaging
  • Fluorescent Dyes - chemistry
  • Image correlation spectroscopy
  • Proteins - chemistry
  • Signal Transduction
  • Spectrometry, Fluorescence - methods
  • TIRF
  • Variance analysis
ispartof: Methods in Enzymology, 2013, Vol.519, p.167-201
description: 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.
language: eng
source: Gale eBooks
identifier: ISSN: 0076-6879
fulltext: fulltext
issn:
  • 0076-6879
  • 1557-7988
url: Link


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descriptionCell–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.
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subjectAggregation state ; Article ; Cell adhesion ; Cell migration ; Cells (Biology) ; CLSM ; Complex stoichiometry ; Fluorescence ; Fluorescence fluctuation ; Fluorescence imaging ; Fluorescent Dyes - chemistry ; Image correlation spectroscopy ; Proteins - chemistry ; Signal Transduction ; Spectrometry, Fluorescence - methods ; TIRF ; Variance analysis
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descriptionCell–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.
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abstractCell–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.
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