Live cell imaging of F-actin dynamics via Fluorescent Speckle Microscopy (FSM)

James Lim, Gaudenz Danuser

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

In this protocol we describe the use of Fluorescent Speckle Microscopy (FSM) to capture high-resolution images of actin dynamics in PtK1 cells. A unique advantage of FSM is its ability to capture the movement and turnover kinetics (assembly/disassembly) of the F-actin network within living cells. This technique is particularly useful in deriving quantitative measurements of F-actin dynamics when paired with computer vision software (qFSM). We describe the selection, microinjection and visualization of fluorescent actin probes in living cells. Importantly, similar procedures are applicable to visualizing other macomolecular assemblies. FSM has been demonstrated for microtubules, intermediate filaments, and adhesion complexes.

Original languageEnglish (US)
Article numbere1325
JournalJournal of Visualized Experiments
Issue number30
DOIs
StatePublished - Aug 2009

Keywords

  • Actin
  • Cellular Biology
  • Cytoskeleton
  • FSM
  • Fluorescence
  • Issue 30
  • Microinjection
  • Microscopy
  • Speckle
  • qFSM

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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