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An improved confocal FRAP technique for the measurement of long‐term actin dynamics in individual stress fibers by J.J. Campbell; M.M. Knight is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Abstract The present study describes an improved fluorescent recovery after photobleaching (FRAP) technique, which has been successfully used to quantify actin dynamics within individual fibers. Chondrocytes were transfected with an eGFP‐actin plasmid and cultured on glass coverslips. In cells expressing eGFP‐actin, confocal microscopy was used to bleach 3 × 1 μm regions accurately positioned along individual stress fibers. The subsequent fluorescent recovery over a 10‐min imaging period was assessed from a series of intensity profiles, positioned along the length of the stress fibers and spanning the bleach region. From these profiles, the normalized fluorescent intensity values were plotted against time. In this way, the technique provided sufficient spatial precision to describe the long‐term behavior within individual stress fibers while accounting for the inherent movement. An identical procedure was used to examine FRAP for eGFP‐actin within the interfiber region. The FRAP curves for stress fibers were accurately modeled by two phase exponentials which indicated only partial recovery with a mobile fraction of 46%. This suggests that some of the F‐actin molecules were in a

Who reads An improved confocal FRAP technique for the measurement of long‐term actin dynamics in individual stress fibers?

Author

J.J. Campbell; M.M. Knight

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 1059-910X)

Published

2007

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)