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

被引：104

作者：

McEvoy, Ann L. ^{[1
]}

Hoi, Hiofan ^{[2
]}

Bates, Mark ^{[3
]}

Platonova, Evgenia ^{[4
]}

Cranfill, Paula J. ^{[5
,6
,10
]}

Baird, Michelle A. ^{[5
,6
,10
]}

Davidson, Michael W. ^{[5
,6
,10
]}

Ewers, Helge ^{[4
]}

Liphardt, Jan ^{[1
,7
,8
,9
]}

Campbell, Robert E. ^{[2
]}

机构：

[1] Univ Calif Berkeley, Biophys Grad Grp, Berkeley, CA 94720 USA

[2] Univ Alberta, Dept Chem, Edmonton, AB, Canada

[3] Max Planck Inst Biophys Chem, Dept NanoBiophoton, D-37077 Gottingen, Germany

[4] Swiss Fed Inst Technol, Inst Biochem, Zurich, Switzerland

[5] Florida State Univ, Natl High Magnet Field Lab, Tallahassee, FL 32306 USA

[6] Florida State Univ, Dept Biol Sci, Tallahassee, FL 32306 USA

[7] Univ Calif Berkeley, Calif Inst Quantitat Biosci QB3, Berkeley, CA 94720 USA

[8] Univ Calif Berkeley, Bay Area Phys Sci Oncol Ctr, Berkeley, CA 94720 USA

[9] Univ Calif Berkeley, Lawrence Berkeley Natl Lab, Phys Biosci Div, Berkeley, CA 94720 USA

[10] Univ Calif Berkeley, Dept Phys, Berkeley, CA 94720 USA

来源：

PLOS ONE | 2012年 / 7卷 / 12期

基金：

美国国家科学基金会; 美国国家卫生研究院; 加拿大自然科学与工程研究理事会;

关键词：

QUANTUM YIELD; RESOLUTION; MICROSCOPY; CELLS; CHEMOTAXIS; EVOLUTION; NANOSCOPY; BRIGHT; GUIDE; LIMIT;

D O I：

10.1371/journal.pone.0051314

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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 pcFP-containing construct.

引用

页数：15

共 54 条

[1] Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein:: structural characterization and applications in fluorescence imaging [J].

Ai, Hui-wang ;

Henderson, J. Nathan ;

Remington, S. James ;

Campbell, Robert E. .

BIOCHEMICAL JOURNAL, 2006, 400 (531-540) :531-540

[2] An optical marker based on the UV-induced green-to-red photoconversion of a fluorescent protein [J].

Ando, R ;

Hama, H ;

Yamamoto-Hino, M ;

Mizuno, H ;

Miyawaki, A .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (20) :12651-12656

[3] Regulated fast nucleocytoplasmic shuttling observed by reversible protein highlighting [J].

Ando, R ;

Mizuno, H ;

Miyawaki, A .

SCIENCE, 2004, 306 (5700) :1370-1373

[4] Photoactivatable Fluorescent Protein mEos2 Displays Repeated Photoactivation after a Long-Lived Dark State in the Red Photoconverted Form [J].

Annibale, P. ;

Scarselli, M. ;

Kodiyan, A. ;

Radenovic, A. .

JOURNAL OF PHYSICAL CHEMISTRY LETTERS, 2010, 1 (09) :1506-1510

[5] Identification of clustering artifacts in photoactivated localization microscopy [J].

Annibale, Paolo ;

Vanni, Stefano ;

Scarselli, Marco ;

Rothlisberger, Ursula ;

Radenovic, Aleksandra .

NATURE METHODS, 2011, 8 (07) :527-528

[6] Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants:: the Keio collection [J].

Baba, Tomoya ;

Ara, Takeshi ;

Hasegawa, Miki ;

Takai, Yuki ;

Okumura, Yoshiko ;

Baba, Miki ;

Datsenko, Kirill A. ;

Tomita, Masaru ;

Wanner, Barry L. ;

Mori, Hirotada .

MOLECULAR SYSTEMS BIOLOGY, 2006, 2 (1) :2006.0008

[7]

Bates M., 2011, Imaging: A Laboratory Manual, P547

[8] Multicolor super-resolution imaging with photo-switchable fluorescent probes [J].

Bates, Mark ;

Huang, Bo ;

Dempsey, Graham T. ;

Zhuang, Xiaowei .

SCIENCE, 2007, 317 (5845) :1749-1753

[9] Imaging intracellular fluorescent proteins at nanometer resolution [J].

Betzig, Eric ;

Patterson, George H. ;

Sougrat, Rachid ;

Lindwasser, O. Wolf ;

Olenych, Scott ;

Bonifacino, Juan S. ;

Davidson, Michael W. ;

Lippincott-Schwartz, Jennifer ;

Hess, Harald F. .

SCIENCE, 2006, 313 (5793) :1642-1645

[10] ABSOLUTE QUANTUM YIELD DETERMINATION BY THERMAL BLOOMING - FLUORESCEIN [J].

BRANNON, JH ;

MAGDE, D .

JOURNAL OF PHYSICAL CHEMISTRY, 1978, 82 (06) :705-709

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