Spectrally Resolved, Functional Super-Resolution Microscopy Reveals Nanoscale Compositional Heterogeneity in Live-Cell Membranes

被引：143

作者：

Moon, Seonah ^{[1
]}

Yan, Rui ^{[1
]}

Kenny, Samuel J. ^{[1
]}

Shyu, Yennie ^{[1
]}

Xiang, Limin ^{[1
]}

Li, Wan ^{[1
]}

Xu, Ke ^{[1
,2
]}

机构：

[1] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA

[2] Lawrence Berkeley Natl Lab, Div Mol Biophys & Integrated Bioimaging, Berkeley, CA 94720 USA

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2017年 / 139卷 / 32期

基金：

美国国家科学基金会;

关键词：

SINGLE-MOLECULE SPECTROSCOPY; LIPID RAFTS; PLASMA-MEMBRANE; CHOLESTEROL CONTENT; FLUORESCENT-PROBES; LIVING CELLS; NILE RED; ORDER; LOCALIZATION; SURFACE;

D O I：

10.1021/jacs.7b03846

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

By recording the full fluorescence spectra and super-resolved positions of similar to 10(6) individual polarity sensing solvatochromic molecules, we reveal compositional heterogeneity in the membranes of live mammalian cells with single-molecule sensitivity and similar to 30 nm spatial resolution. This allowed us to unveil distinct polarity characteristics of the plasma membrane and the membranes of nanoscale intracellular organelles, a result we found to be due to differences in cholesterol levels. Within the plasma membrane, we observed the formation of low polarity, raft-like nanodomains upon cholesterol addition or cholera-toxin treatment, but found this nanoscale phase separation absent in native cells. The ultimate sensitivity achieved through examining the spectra of individual molecules thus opens the door to functional interrogations of intracellular physicochemical parameters at the nano scale.

引用

页码：10944 / 10947

页数：4

共 39 条

[1] Multi-dimensional super-resolution imaging enables surface hydrophobicity mapping [J].

Bongiovanni, Marie N. ;

Godet, Julien ;

Horrocks, Mathew H. ;

Tosatto, Laura ;

Carr, Alexander R. ;

Wirthensohn, David C. ;

Ranasinghe, Rohan T. ;

Lee, Ji-Eun ;

Ponjavic, Aleks ;

Fritz, Joelle V. ;

Dobson, Christopher M. ;

Klenerman, David ;

Lee, Steven F. .

Nature Communications, 2016, 7

[2] Live Intracellular Super-Resolution Imaging Using Site-Specific Stains [J].

Carlini, Lina ;

Manley, Suliana .

ACS CHEMICAL BIOLOGY, 2013, 8 (12) :2643-2648

[3] Monitoring Biophysical Properties of Lipid Membranes by Environment-Sensitive Fluorescent Probes [J].

Demchenko, Alexander P. ;

Mely, Yves ;

Duportail, Guy ;

Klymchenko, Andrey S. .

BIOPHYSICAL JOURNAL, 2009, 96 (09) :3461-3470

[4] Super-resolution spectroscopic microscopy via photon localization [J].

Dong, Biqin ;

Almassalha, Luay ;

Urban, Ben E. ;

The-Quyen Nguyen ;

Khuon, Satya ;

Chew, Teng-Leong ;

Backman, Vadim ;

Sun, Cheng ;

Zhang, Hao F. .

NATURE COMMUNICATIONS, 2016, 7

[5] Laurdan Fluorescence Lifetime Discriminates Cholesterol Content from Changes in Fluidity in Living Cell Membranes [J].

Golfetto, Ottavia ;

Hinde, Elizabeth ;

Gratton, Enrico .

BIOPHYSICAL JOURNAL, 2013, 104 (06) :1238-1247

[6]

GREENSPAN P, 1985, J LIPID RES, V26, P781

[7] Correlative Super-Resolution Microscopy: New Dimensions and New Opportunities [J].

Hauser, Meghan ;

Wojcik, Michal ;

Kim, Doory ;

Mahmoudi, Morteza ;

Li, Wan ;

Xu, Ke .

CHEMICAL REVIEWS, 2017, 117 (11) :7428-7456

[8]

Hell, 2007, SCIENCE, V316, P1153

[9] Lipid traffic: Floppy drives and a superhighway [J].

Holthuis, JCM ;

Levine, TP .

NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2005, 6 (03) :209-220

[10] Lipid landscapes and pipelines in membrane homeostasis [J].

Holthuis, Joost C. M. ;

Menon, Anant K. .

NATURE, 2014, 510 (7503) :48-57

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