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Title: Supercontinuum Stimulated Emission Depletion Fluorescence Lifetime Imaging

Abstract

Supercontinuum (SC) stimulated emission depletion (STED) fluorescence lifetime imaging is demonstrated by using time-correlated single-photon counting (TCSPC) detection. The spatial resolution of the developed STED instrument was measured by imaging monodispersed 40-nm fluorescent beads and then determining their fwhm, and was 36 ± 9 and 40 ± 10 nm in the X and Y coordinates, respectively. The same beads measured by confocal microscopy were 450 ± 50 and 430 ± 30 nm, which is larger than the diffraction limit of light due to underfilling the microscope objective. Underfilling the objective and time gating the signal were necessary to achieve the stated STED spatial resolution. The same fluorescence lifetime (2.0 ± 0.1 ns) was measured for the fluorescent beads by using confocal or STED lifetime imaging. The instrument has been applied to study Alexa Fluor 594-phalloidin labeled F-actin-rich projections with dimensions smaller than the diffraction limit of light in cultured cells. Fluorescence lifetimes of the actin-rich projections range from 2.2 to 2.9 ns as measured by STED lifetime imaging.

Authors:
; ; ;
Publication Date:
Research Org.:
Ames Laboratory (AMES), Ames, IA (United States)
Sponsoring Org.:
USDOE SC Office of Advanced Scientific Computing Research (SC-21)
OSTI Identifier:
1044798
Report Number(s):
IS-J 7676
DOE Contract Number:  
DE-AC02-07CH11358
Resource Type:
Journal Article
Journal Name:
Journal of Physical Chemistry B
Additional Journal Information:
Journal Volume: June
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE

Citation Formats

Lesoine, Michael, Bose, Sayantan, Petrich, Jacob, and Smith, Emily. Supercontinuum Stimulated Emission Depletion Fluorescence Lifetime Imaging. United States: N. p., 2012. Web. doi:10.1021/jp303912p.
Lesoine, Michael, Bose, Sayantan, Petrich, Jacob, & Smith, Emily. Supercontinuum Stimulated Emission Depletion Fluorescence Lifetime Imaging. United States. doi:10.1021/jp303912p.
Lesoine, Michael, Bose, Sayantan, Petrich, Jacob, and Smith, Emily. Wed . "Supercontinuum Stimulated Emission Depletion Fluorescence Lifetime Imaging". United States. doi:10.1021/jp303912p.
@article{osti_1044798,
title = {Supercontinuum Stimulated Emission Depletion Fluorescence Lifetime Imaging},
author = {Lesoine, Michael and Bose, Sayantan and Petrich, Jacob and Smith, Emily},
abstractNote = {Supercontinuum (SC) stimulated emission depletion (STED) fluorescence lifetime imaging is demonstrated by using time-correlated single-photon counting (TCSPC) detection. The spatial resolution of the developed STED instrument was measured by imaging monodispersed 40-nm fluorescent beads and then determining their fwhm, and was 36 ± 9 and 40 ± 10 nm in the X and Y coordinates, respectively. The same beads measured by confocal microscopy were 450 ± 50 and 430 ± 30 nm, which is larger than the diffraction limit of light due to underfilling the microscope objective. Underfilling the objective and time gating the signal were necessary to achieve the stated STED spatial resolution. The same fluorescence lifetime (2.0 ± 0.1 ns) was measured for the fluorescent beads by using confocal or STED lifetime imaging. The instrument has been applied to study Alexa Fluor 594-phalloidin labeled F-actin-rich projections with dimensions smaller than the diffraction limit of light in cultured cells. Fluorescence lifetimes of the actin-rich projections range from 2.2 to 2.9 ns as measured by STED lifetime imaging.},
doi = {10.1021/jp303912p},
journal = {Journal of Physical Chemistry B},
number = ,
volume = June,
place = {United States},
year = {2012},
month = {6}
}