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TitleEnhancing fluorescent protein photostability through robot-assisted photobleaching.
Publication TypeJournal Article
Year of Publication2018
AuthorsWiens, M D., F Hoffmann, Y Chen, and R E. Campbell
JournalIntegr Biol (Camb)
Volume10
Issue7
Pagination419-428
Date Published2018 Jul 16
ISSN1757-9708
Abstract

Improving fluorescent proteins through the use of directed evolution requires robust techniques for screening large libraries of genetic variants. Here we describe an effective and relatively low-cost system for screening libraries of fluorescent protein variants for improved photostability in the context of colonies on a Petri dish. Application of this system to the yellow fluorescent protein mCitrine, led to the development of Citrine2 with improved photostability and similar high fluorescent brightness. The photobleaching robot was constructed using a Lego Mindstorms Ev3 set and a xenon arc lamp, which together create even and high irradiance over an entire Petri dish through patterned illumination.

DOI10.1039/c8ib00063h
Alternate JournalIntegr Biol (Camb)
PubMed ID29897363
PubMed Central IDPMC6047911
Grant ListU01 NS090565 / NS / NINDS NIH HHS / United States
U01 NS094246 / NS / NINDS NIH HHS / United States