TY - JOUR
T1 - Guide to red fluorescent proteins and biosensors for flow cytometry
AU - Piatkevich, Kiryl D.
AU - Verkhusha, Vladislav V.
N1 - Funding Information:
For helpful discussions, the authors thank Drs. Jinghang Zhang and Steven Porcelli of the Einstein Flow Cytometry Core Facility (supported by the Einstein Cancer Center (NIH/NCI grant CA013330) and the Einstein Center for AIDS Research (NIH grant AI-51519)). This work was supported by the NIH/NIGMS grant GM073913 to V.V.V.
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
KW - Biosensor
KW - Molecular brightness
KW - Monomerization
KW - Multicolor flow cytometry
KW - Oligomerization
KW - Red fluorescent protein
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U2 - 10.1016/B978-0-12-374912-3.00017-1
DO - 10.1016/B978-0-12-374912-3.00017-1
M3 - Article
C2 - 21704849
AN - SCOPUS:79959598821
SN - 0091-679X
VL - 102
SP - 431
EP - 461
JO - Methods in cell biology
JF - Methods in cell biology
ER -