FRET microscopy to visualize transcription factor dimerization in the nucleus of the living cell

Richard N. Day; Ammasi Periasamy

doi:10.1117/12.424550

24 April 2001 FRET microscopy to visualize transcription factor dimerization in the nucleus of the living cell

Richard N. Day, Ammasi Periasamy

Proceedings Volume 4262, Multiphoton Microscopy in the Biomedical Sciences; (2001) https://doi.org/10.1117/12.424550
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

Cells respond to environmental cues or developmental programs by modifying protein complexes in the nucleus to alter patterns of gene transcription. Recent advances in digital imaging coupled with the development of new fluorescent probes provide the tools to begin to study where and when changes in protein interactions take place in the nucleus of the living cell. Here, we describe the application of fluorescence resonance energy transfer (FRET) using both wide-field and 2-photon (2P) microscopy to visualize the interactions of the transcription factor CAATT/enhancer binding protein alpha (C/EBP(alpha) ) in living pituitary cells. The efficiency of FRET will be improved if the overlap of the donor emission spectra with the absorption spectra for the acceptor is increased. The trade off for this improved efficiency, however, is that there will be an increase in the background signal from which the weak sensitized acceptor emission must be extracted. Here, we compare and contrast the FRET signals obtained from dimerized C/EBP(alpha) proteins fused to several different color variants of the jellyfish green fluorescent protein (GFP). We use both wide-field and 2P FRET microscopy to characterize the spectral cross-talk and FRET signals for each of the donor and acceptor pairs.

Citation Download Citation

Richard N. Day and Ammasi Periasamy "FRET microscopy to visualize transcription factor dimerization in the nucleus of the living cell", Proc. SPIE 4262, Multiphoton Microscopy in the Biomedical Sciences, (24 April 2001); https://doi.org/10.1117/12.424550

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
8 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Fluorescence resonance energy transfer

Proteins

Microscopy

Green fluorescent protein

Luminescence

Fluorescent proteins

Absorption

Show All Keywords

Keywords/Phrases

Search In:

Publication Years