Video-rate Confocal Microscopy for Single-molecule Imaging in Live Cells and Superresolution Fluorescence Imaging

Overview

Journal Biophys J

Publisher Cell Press

Specialty Biophysics

Date 2012 Oct 23

PMID 23083712

Citations 17

Authors

Jinwoo Lee

Yukihiro Miyanaga

Masahiro Ueda

Sungchul Hohng

Affiliations

Soon will be listed here.

Abstract

There is no confocal microscope optimized for single-molecule imaging in live cells and superresolution fluorescence imaging. By combining the swiftness of the line-scanning method and the high sensitivity of wide-field detection, we have developed a, to our knowledge, novel confocal fluorescence microscope with a good optical-sectioning capability (1.0 μm), fast frame rates (<33 fps), and superior fluorescence detection efficiency. Full compatibility of the microscope with conventional cell-imaging techniques allowed us to do single-molecule imaging with a great ease at arbitrary depths of live cells. With the new microscope, we monitored diffusion motion of fluorescently labeled cAMP receptors of Dictyostelium discoideum at both the basal and apical surfaces and obtained superresolution fluorescence images of microtubules of COS-7 cells at depths in the range 0-85 μm from the surface of a coverglass.

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