ArticleScientific reports2015
Two-photon directed evolution of green fluorescent proteins.
Article in Scientific reports, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 29 citations in OpenAlex.
- Active pixel power control for crosstalk-free all-optical neural interrogation.Nature communications · 2026Article
- Orthogonal RNA replication enables directed evolution and Darwinian adaptation in mammalian cells.Nature chemical biology · 2025Article
- Next-generation cell lines for profiling different proteasome forms and their implications in cancer.Frontiers in immunology · 2025Article
- Longitudinal tracking of acute kidney injury reveals injury propagation along the nephron.Nature communications · 2023Article
- Impact of the Protein Environment on Two-Photon Absorption Cross-Sections of the GFP Chromophore Anion Resolved at the XMCQDPT2 Level of Theory.International journal of molecular sciences · 2023Article
- FRET Imaging of Rho GTPase Activity with Red Fluorescent Protein-Based FRET Pairs.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Recent advances in nonlinear microscopy: Deep insights and polarized revelations.The international journal of biochemistry & cell biology · 2021Review
- High throughput instrument to screen fluorescent proteins under two-photon excitation.Biomedical optics express · 2020Article
- ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems.BMC biology · 2020Article
- CytoMAP: A Spatial Analysis Toolbox Reveals Features of Myeloid Cell Organization in Lymphoid Tissues.Cell reports · 2020Article
- Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than Enhanced Green Fluorescent Protein under Two-Photon Excitation.The journal of physical chemistry letters · 2017Article
- Deciphering the molecular mechanism responsible for GCaMP6m's Ca2+-dependent change in fluorescence.PloS one · 2017Article
- Optimizing ultrafast illumination for multiphoton-excited fluorescence imaging.Biomedical optics express · 2016Article
- High contrast two-photon imaging of fingermarks.Scientific reports · 2016Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 2 countries.
Funding
Abstract
Directed evolution has been used extensively to improve the properties of a variety of fluorescent proteins (FPs). Evolutionary strategies, however, have not yet been used to improve the two-photon absorption (2PA) properties of a fluorescent protein, properties that are important for two-photon imaging in living tissues, including the brain. Here we demonstrate a technique for quantitatively screening the two-photon excited fluorescence (2PEF) efficiency and 2PA cross section of tens of thousands of mutant FPs expressed in E. coli colonies. We use this procedure to move EGFP through three rounds of two-photon directed evolution leading to new variants showing up to a 50% enhancement in peak 2PA cross section and brightness within the near-IR tissue transparency wavelength range.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.