Quantitative imaging of protein interactions in the cell nucleus

Ty C. Voss, Ignacio A. Demarco, Richard Day

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Over the past decade, genetically encoded fluorescent proteins have become widely used as noninvasive markers in living cells. The development of fluorescent proteins, coupled with advances in digital imaging, has led to the rapid evolution of live-cell imaging methods. These approaches are being applied to address biological questions of the recruitment, co-localization, and interactions of specific proteins within particular subcellular compartments. In the wake of this rapid progress, however, come important issues associated with the acquisition and analysis of ever larger and more complex digital imaging data sets. Using protein localization in the mammalian cell nucleus as an example, we will review some recent developments In the application of quantitative imaging to analyze subcellular distribution and co-localization of proteins in populations of living cells. In this report, we review the principles of acquiring fluorescence resonance energy transfer (FRET) microscopy measurements to define the spatial relationships between proteins. We then discuss how fluorescence lifetime imaging microscopy (FLIM) provides a method that is independent of intensity-based measurements to detect localized protein interactions with spatial resolution. Finally, we consider potential problems associated with the expression of proteins fused to fluorescent proteins for FRET-based measurements from living cells.

Original languageEnglish (US)
Pages (from-to)413-424
Number of pages12
JournalBioTechniques
Volume38
Issue number3
StatePublished - Mar 2005
Externally publishedYes

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Cell Nucleus
Cells
Imaging techniques
Proteins
Fluorescence Resonance Energy Transfer
Microscopy
Microscopic examination
Optical Imaging
Fluorescence
Population

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Quantitative imaging of protein interactions in the cell nucleus. / Voss, Ty C.; Demarco, Ignacio A.; Day, Richard.

In: BioTechniques, Vol. 38, No. 3, 03.2005, p. 413-424.

Research output: Contribution to journalArticle

Voss, TC, Demarco, IA & Day, R 2005, 'Quantitative imaging of protein interactions in the cell nucleus', BioTechniques, vol. 38, no. 3, pp. 413-424.
Voss, Ty C. ; Demarco, Ignacio A. ; Day, Richard. / Quantitative imaging of protein interactions in the cell nucleus. In: BioTechniques. 2005 ; Vol. 38, No. 3. pp. 413-424.
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