Guide to red fluorescent proteins and biosensors for flow cytometry

Kiryl D. Piatkevich, Vladislav Verkhusha

Research output: Book/ReportBook

28 Citations (Scopus)

Abstract

Since the discovery of the first red fluorescent protein (RFP), named DsRed, 12 years ago, a wide pallet of red-shifted fluorescent proteins has been cloned and biotechnologically developed into monomeric fluorescent probes for optical microscopy. Several new types of monomeric RFPs that change the emission wavelength either with time, called fluorescent timers, or after a brief irradiation with violet light, known as photoactivatable proteins, have been also engineered. Moreover, RFPs with a large Stokes shift of fluorescence emission have been recently designed. Because of their distinctive excitation and fluorescence detection conditions developed specifically for microscopy, these fluorescent probes can be suboptimal for flow cytometry. Here, we have selected and summarized the advanced orange, red, and far-red fluorescent proteins with the properties specifically required for the flow cytometry applications. Their effective brightness was calculated for the laser sources available for the commercial flow cytometers and sorters. Compatibility of the fluorescent proteins of different colors in a multiparameter flow cytometry was determined. Novel FRET pairs, utilizing RFPs, RFP-based intracellular biosensors, and their application to a high-throughput screening, are also discussed.

Original languageEnglish (US)
PublisherUnknown Publisher
Number of pages31
Volume102
DOIs
StatePublished - 2011

Publication series

NameMethods in Cell Biology
Volume102
ISSN (Print)0091679X

Fingerprint

Biosensing Techniques
Flow Cytometry
Fluorescent Dyes
Microscopy
Fluorescence
Proteins
Lasers
Color
Light
red fluorescent protein

Keywords

  • Biosensor
  • Molecular brightness
  • Monomerization
  • Multicolor flow cytometry
  • Oligomerization
  • Red fluorescent protein

ASJC Scopus subject areas

  • Cell Biology

Cite this

Guide to red fluorescent proteins and biosensors for flow cytometry. / Piatkevich, Kiryl D.; Verkhusha, Vladislav.

Unknown Publisher, 2011. 31 p. (Methods in Cell Biology; Vol. 102).

Research output: Book/ReportBook

Piatkevich, Kiryl D. ; Verkhusha, Vladislav. / Guide to red fluorescent proteins and biosensors for flow cytometry. Unknown Publisher, 2011. 31 p. (Methods in Cell Biology).
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