Green-to-Red Photoconversion of GCaMP

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Genetically encoded calcium indicators (GECIs) permit imaging intracellular calcium transients. Among GECIs, the GFP-based GCaMPs are the most widely used because of their high sensitivity and rapid response to changes in intracellular calcium concentrations. Here we report that the fluorescence of GCaMPs-including GCaMP3, GCaMP5 and GCaMP6-can be converted from green to red following exposure to blue-green light (450500 nm). This photoconversion occurs in both insect and mammalian cells and is enhanced in a low oxygen environment. The red fluorescent GCaMPs retained calcium responsiveness, albeit with reduced sensitivity. We identified several amino acid residues in GCaMP important for photoconversion and generated a GCaMP variant with increased photoconversion efficiency in cell culture. This light-induced spectral shift allows the ready labeling of specific, targeted sets of GCaMP-expressing cells for functional imaging in the red channel. Together, these findings indicate the potential for greater utility of existing GCaMP reagents, including transgenic animals
Publisher
PUBLIC LIBRARY SCIENCE
Issue Date
2015-09
Language
English
Article Type
Article
Keywords

ENCODED CALCIUM INDICATORS; FLUORESCENT PROTEIN; NEURAL ACTIVITY; CA2+ INDICATORS; PHOTOACTIVATION; CONVERSION; MECHANISM; TRACKING; LIGHT; CELL

Citation

PLOS ONE, v.10, no.9

ISSN
1932-6203
DOI
10.1371/journal.pone.0138127
URI
http://hdl.handle.net/10203/211878
Appears in Collection
BS-Journal Papers(저널논문)
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