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

Richard Day, A. Periasamy

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

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 steady-state and two-photon excitation (TPE) microscopy to visualize the interactions of the transcription factor CAATT/enhancer binding protein alpha (C/EBPα) 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/EBPα 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.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsA. Periasamy, P.T.C. So
Pages163-170
Number of pages8
Volume4262
DOIs
StatePublished - 2001
Externally publishedYes
EventMultiphoton Microscopy in the Biomedical Sciences - San Jose, CA, United States
Duration: Jan 21 2001Jan 23 2001

Other

OtherMultiphoton Microscopy in the Biomedical Sciences
CountryUnited States
CitySan Jose, CA
Period1/21/011/23/01

Fingerprint

Transcription factors
Dimerization
resonance fluorescence
dimerization
Microscopic examination
energy transfer
Cells
microscopy
proteins
nuclei
Proteins
Transcription
cues
Absorption spectra
cells
genes
Photons
Genes
Fluorescence Resonance Energy Transfer
Color

Keywords

  • Colocalization
  • Fluorescence resonance energy transfer (FRET) microscopy
  • Green fluorescent protein
  • Transcription factors
  • Two-photon excitation

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

Cite this

Day, R., & Periasamy, A. (2001). FRET microscopy to visualize transcription factor dimerization in the nucleus of the living cell. In A. Periasamy, & P. T. C. So (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 4262, pp. 163-170) https://doi.org/10.1117/12.424550

FRET microscopy to visualize transcription factor dimerization in the nucleus of the living cell. / Day, Richard; Periasamy, A.

Proceedings of SPIE - The International Society for Optical Engineering. ed. / A. Periasamy; P.T.C. So. Vol. 4262 2001. p. 163-170.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Day, R & Periasamy, A 2001, FRET microscopy to visualize transcription factor dimerization in the nucleus of the living cell. in A Periasamy & PTC So (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 4262, pp. 163-170, Multiphoton Microscopy in the Biomedical Sciences, San Jose, CA, United States, 1/21/01. https://doi.org/10.1117/12.424550
Day R, Periasamy A. FRET microscopy to visualize transcription factor dimerization in the nucleus of the living cell. In Periasamy A, So PTC, editors, Proceedings of SPIE - The International Society for Optical Engineering. Vol. 4262. 2001. p. 163-170 https://doi.org/10.1117/12.424550
Day, Richard ; Periasamy, A. / FRET microscopy to visualize transcription factor dimerization in the nucleus of the living cell. Proceedings of SPIE - The International Society for Optical Engineering. editor / A. Periasamy ; P.T.C. So. Vol. 4262 2001. pp. 163-170
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