Publication details

A Surprising Photoactivity of Blue Fluorescent Protein TagBFP Allows for Super-Resolution Microscopy

Authors

KLEMENTIEVA N.V. LUKYANOV K.A. GORBACHEV D.A. CHUDAKOV Dmitriy ZAGAYNOVA E.V. MISHIN A.S.

Year of publication 2018
Type Article in Periodical
Magazine / Source SOVREMENNYE TEHNOLOGII V MEDICINE
MU Faculty or unit

Central European Institute of Technology

Citation
web https://journals.aps.org/prl/abstract/10.1103/PhysRevLett.120.233901
Doi http://dx.doi.org/10.17691/stm2018.10.1.04
Keywords TagBFP; photoactivation; blue-to-red photoconversion; blinking; super-resolution microscopy
Description The aim of this research was to study photobehavior of a popular blue fluorescent protein TagBFP and apply this marker for super-resolution microscopy. Materials and Methods. Photoactivation of TagBFP was examined both in protein solution in vitro and in living cells. Subdiffraction imaging was performed using total internal reflection fluorescence microscopy followed by super-resolution radial fluctuations or single-molecule localization analysis. Results. We show that TagBFP exhibits blinking behavior upon 405 nm light illumination. Moreover, photoactivation to red-emitting state is occurring in the conditions typically used for TagBFP imaging. The red (photoactivated) form of TagBFP possesses spectral properties similar to TagRFP - a close homologue of TagBFP. We show that both blinking and photoactivation of TagBFP can be utilized for super-resolution imaging. We conclude that photoactivation of TagBFP to red-emitting form should be taken into account in the design of multi-channel imaging experiments involving high-power or prolonged UV illumination.

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