Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.3/156034

Title

Fixation alters fluorescence lifetime and anisotropy of cells expressing EYFP-tagged serotonin 1A receptor

Author(s)

Ganguly, Sourav;  Clayton, Andrew H. A.;  Chattopadhyay, Amitabha

Abstract

Fluorescence microscopic approaches represent powerful techniques to monitor molecular interactions in the cellular milieu. Measurements of fluorescence lifetime and anisotropy enjoy considerable popularity in this context. These measurements are often performed on live as well as fixed cells. We report here that formaldehyde-induced cell fixation introduces heterogeneities in the fluorescence emission of serotonin 1A receptors tagged to enhanced yellow fluorescent protein, and alters fluorescence lifetime and anisotropy significantly. To the best of our knowledge, our results constitute the first report on the effect of formaldehyde fixation on fluorescence parameters of cellular proteins. We conclude that fluorescence parameters derived from fixed cells should be interpreted with caution.

Publication type

Journal article

Research centre

Swinburne University of Technology. Faculty of Engineering and Industrial Sciences. Centre for Micro-Photonics

Source

Biochemical and Biophysical Research Communications, Vol. 405, no. 2 (Feb 2011), pp. 234-237

Publication year

2011

FOR Code(s)

0304 Medicinal and Biomolecular Chemistry;  0601 Biochemistry and Cell Biology;  1101 Medical Biochemistry and Metabolomics

Keyword(s)

Anisotropy;  Fixation;  FLIM;  Flourescence lifetime;  Fluorescence lifetime imaging microscopy;  Fluorescence resonance energy transfer;  FRET

Publisher

Elsevier

ISSN

0006-291X

Publisher URL

http://dx.doi.org/10.1016/j.bbrc.2011.01.016

Copyright

Copyright © 2011 Elsevier Inc. All rights reserved.

Peer reviewed