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Spectrally Resolved, Functional Super-Resolution Microscopy Reveals Nanoscale Compositional Heterogeneity in Live-Cell Membranes.
- Source :
-
Journal of the American Chemical Society [J Am Chem Soc] 2017 Aug 16; Vol. 139 (32), pp. 10944-10947. Date of Electronic Publication: 2017 Aug 07. - Publication Year :
- 2017
-
Abstract
- By recording the full fluorescence spectra and super-resolved positions of ∼10 <superscript>6</superscript> individual polarity-sensing solvatochromic molecules, we reveal compositional heterogeneity in the membranes of live mammalian cells with single-molecule sensitivity and ∼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 nanoscale.
- Subjects :
- Animals
COS Cells
Cell Line
Cell Membrane chemistry
Cell Survival
Chlorocebus aethiops
Endoplasmic Reticulum chemistry
Fluorescent Dyes analysis
Microscopy, Fluorescence methods
Mitochondrial Membranes chemistry
Oxazines analysis
Potoroidae
Cell Membrane ultrastructure
Cholesterol analysis
Endoplasmic Reticulum ultrastructure
Mitochondrial Membranes ultrastructure
Spectrometry, Fluorescence methods
Subjects
Details
- Language :
- English
- ISSN :
- 1520-5126
- Volume :
- 139
- Issue :
- 32
- Database :
- MEDLINE
- Journal :
- Journal of the American Chemical Society
- Publication Type :
- Academic Journal
- Accession number :
- 28774176
- Full Text :
- https://doi.org/10.1021/jacs.7b03846