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Updated: May 18, 2026
19:16
Live Cell Imaging of F-actin Dynamics via Fluorescent Speckle Microscopy (FSM)
Published on: August 5, 2009
Quantitative fluorescent speckle microscopy (QFSM) to measure actin dynamics
Michelle C Mendoza1, Sebastien Besson, Gaudenz Danuser
1Department of Cell Biology, Harvard Medical School, Boston, MA, USA.
Current Protocols in Cytometry
|October 9, 2012
Summary
Quantitative fluorescent speckle microscopy (QFSM) analyzes macromolecular dynamics in living cells. This method tracks actin filament networks to reveal cell migration mechanisms.
Area of Science:
- Cell Biology
- Biophysics
- Microscopy
Background:
- Quantitative fluorescent speckle microscopy (QFSM) is a powerful live-cell imaging technique.
- It offers high spatial and temporal resolution for analyzing macromolecular assembly dynamics.
- Previous successes include studies on actin filament dynamics during cell migration and microtubule dynamics.
Purpose of the Study:
- To illustrate the procedures of FSM imaging and computational image processing for QFSM.
- To focus on the application of QFSM in analyzing actin filament dynamics.
- To demonstrate how QFSM extracts quantitative information about cellular processes.
Main Methods:
- Microinjection of fluorophore-labeled actin into cells.
- Live-cell imaging using spinning disk confocal or wide-field microscopy.
- Computational tracking of fluorescent speckles (actin monomer clusters) to derive network dynamics.
Main Results:
- QFSM measures movement and turnover kinetics of the entire F-actin network in living cells.
- Speckle tracking allows derivation of network flows.
- Maps of filament assembly and disassembly rates are generated.
Conclusions:
- QFSM provides a comprehensive method for analyzing F-actin network dynamics in real-time.
- The technique offers advantages over other methods by analyzing the entire field of view.
- QFSM is valuable for understanding cell migration and other dynamic cellular processes.
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