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mMaple: A Photoconvertible Fluorescent Protein for Use in Multiple Imaging Modalities

Journal Article · · PLoS ONE
 [1];  [2];  [3];  [4];  [5];  [5];  [5];  [4];  [6];  [2];
  1. Univ. of California, Berkeley, CA (United States). Biophysics Graduate Group; DOE/OSTI
  2. Univ. of Alberta, Edmonton, AB (Canada). Dept. of Chemistry
  3. Max Planck Inst. for Biophysical Chemistry, Gottingen (Germany). Dept. of NanoBiophotonics
  4. Eidgenossische Technische Hochschule (ETH) Zurich, (Switzerland). Inst. of Biochemistry
  5. Florida State Univ., Tallahassee, FL (United States). National High Magnetic Field Lab. (MagLab); Univ. of California, Berkeley, CA (United States). Dept. of Physics
  6. Univ. of California, Berkeley, CA (United States). Biophysics Graduate Group; Univ. of California, Berkeley, CA (United States). California Inst. for Quantitative Biosciences (QB3); Univ. of California, Berkeley, CA (United States). Bay Area Physical Sciences. Oncology Center; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Physical Biosciences Division

Recent advances in fluorescence microscopy have extended the spatial resolution to the nanometer scale. Here, we report an engineered photoconvertible fluorescent protein (pcFP) variant, designated as mMaple, that is suited for use in multiple conventional and super-resolution imaging modalities, specifically, widefield and confocal microscopy, structured illumination microscopy (SIM), and single-molecule localization microscopy. We demonstrate the versatility of mMaple by obtaining super-resolution images of protein organization in Escherichia coli and conventional fluorescence images of mammalian cells. Beneficial features of mMaple include high photostability of the green state when expressed in mammalian cells and high steady state intracellular protein concentration of functional protein when expressed in E. coli. mMaple thus enables both fast live-cell ensemble imaging and high precision single molecule localization for a single pcFPcontaining construct.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1627567
Journal Information:
PLoS ONE, Journal Name: PLoS ONE Journal Issue: 12 Vol. 7; ISSN 1932-6203
Publisher:
Public Library of ScienceCopyright Statement
Country of Publication:
United States
Language:
English

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Cited By (30)

Progress in quantitative single-molecule localization microscopy journal April 2014
Challenges in quantitative single molecule localization microscopy journal June 2014
Nuclear pores as versatile reference standards for quantitative superresolution microscopy journal September 2019
Monomerization of the photoconvertible fluorescent protein SAASoti by rational mutagenesis of single amino acids journal October 2018
Dynamics and distribution of paxillin, vinculin, zyxin and VASP depend on focal adhesion location and orientation journal July 2019
Single-molecule fluorescence microscopy review: shedding new light on old problems journal July 2017
Super-resolution binding activated localization microscopy through reversible change of DNA conformation journal March 2018
Synthetic and genetic dimers as quantification ruler for single-molecule counting with PALM journal June 2019
Systematic analysis of the molecular architecture of endocytosis reveals a nanoscale actin nucleation template that drives efficient vesicle formation journal November 2017
Recruitment, Assembly, and Molecular Architecture of the SpoIIIE DNA Pump Revealed by Superresolution Microscopy journal May 2013
Super-Resolution Imaging of Bacteria in a Microfluidics Device journal October 2013
In cellulo Evaluation of Phototransformation Quantum Yields in Fluorescent Proteins Used As Markers for Single-Molecule Localization Microscopy journal June 2014
Green-to-Red Photoconversion of GCaMP journal September 2015
Super-Resolution Molecular and Functional Imaging of Nanoscale Architectures in Life and Materials Science journal June 2014
Genetically encoded Ca2+ indicators; expanded affinity range, color hue and compatibility with optogenetics journal November 2014
Photoconvertible Fluorescent Proteins and the Role of Dynamics in Protein Evolution journal August 2017
Novel Phototransformable Fluorescent Protein SAASoti with Unique Photochemical Properties journal July 2019
Frequent exchange of the DNA polymerase during bacterial chromosome replication journal March 2017
Induction of Signal Transduction by Using Non-Channelrhodopsin-Type Optogenetic Tools journal May 2018
Primed Conversion: The New Kid on the Block for Photoconversion journal May 2018
Single-molecule evaluation of fluorescent protein photoactivation efficiency using an in vivo nanotemplate journal January 2014
Optogenetic control with a photocleavable protein, PhoCl journal March 2017
Fluorescent proteins for live-cell imaging with super-resolution journal January 2014
Characterization and development of photoactivatable fluorescent proteins for single-molecule-based superresolution imaging journal May 2014
Replisome activity slowdown after exposure to ultraviolet light in Escherichia coli journal May 2019
Topological data analysis quantifies biological nano-structure from single molecule localization microscopy journal October 2019
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Nuclear pores as versatile reference standards for quantitative superresolution microscopy journal January 2019
A cost-efficient open source laser engine for microscopy journal January 2019
Cost-efficient open source laser engine for microscopy journal January 2020

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