Improving the photostability of bright monomeric orange and red fluorescent proteins.

ABSTRACT: All organic fluorophores undergo irreversible photobleaching during prolonged illumination. Although fluorescent proteins typically bleach at a substantially slower rate than many small-molecule dyes, in many cases the lack of sufficient photostability remains an important limiting factor for experiments requiring large numbers of images of single cells. Screening methods focusing solely on brightness or wavelength are highly effective in optimizing both properties, but the absence of selective pressure for photostability in such screens leads to unpredictable photobleaching behavior in the resulting fluorescent proteins. Here we describe an assay for screening libraries of fluorescent proteins for enhanced photostability. With this assay, we developed highly photostable variants of mOrang

SUBMITTER: Shaner NC 

PROVIDER: S-EPMC2853173 | biostudies-literature | 2008 Jun

REPOSITORIES: biostudies-literature