Fluorescence lifetime imaging by asynchronous pump-probe microscopy

Biophysical Journal
C Y DongE Gratton

Abstract

We report the development of a scanning lifetime fluorescence microscope using the asynchronous, pump-probe (stimulated emission) approach. There are two significant advantages of this technique. First, the cross-correlation signal produced by overlapping the pump and probe lasers results in i) an axial sectioning effect similar to that in confocal and two-photon excitation microscopy, and ii) improved spatial resolution compared to conventional one-photon fluorescence microscopy. Second, the low-frequency, cross-correlation signal generated allows lifetime-resolved imaging without using fast photodetectors. The data presented here include 1) determination of laser sources' threshold powers for linearity in the pump-probe signal; 2) characterization of the pump-probe intensity profile using 0.28 microns fluorescent latex spheres; 3) high frequency (up to 6.7 GHz) lifetime measurement of rhodamine B in water; and 4) lifetime-resolved images of fluorescent latex spheres, human erythrocytes and a mouse fibroblast cell stained by rhodamine DHPE, and a mouse fibroblast labeled with ethidium bromide and rhodamine DHPE.

Citations

May 21, 1997·Biophysical Chemistry·I GryczynskiJ R Lakowicz
Feb 11, 2009·Nature Biotechnology·Enrico Gratton, Michelle Digman
Feb 27, 1999·Journal of Microscopy·A Squire, P I Bastiaens
Jul 19, 2000·Proceedings of the National Academy of Sciences of the United States of America·T A KlarS W Hell
Jan 28, 2009·Photochemistry and Photobiology·Tong YeWarren S Warren
Feb 23, 2012·Journal of Biomedical Optics·Thilo DellwigFu-Jen Kao
Apr 2, 2011·Annual Review of Physical Chemistry·Wei MinX Sunney Xie
Nov 1, 2010·Expert Opinion on Medical Diagnostics·Chen Yuan Dong, Paul J Campagnola
Nov 8, 2011·Current Opinion in Chemical Biology·Wei Min
Apr 3, 2016·The Review of Scientific Instruments·Martin C FischerWarren S Warren
Mar 29, 2000·Biophysical Journal·P J VerveerP I Bastiaens
Oct 6, 1999·Current Opinion in Structural Biology·M G Gustafsson
Sep 20, 2005·Biophysical Journal·Alessandro EspositoFred S Wouters
Jan 30, 2008·Current Protocols in Cell Biology·Alessandro Esposito, Fred S Wouters
Oct 1, 1996·Photochemistry and Photobiology·J R LakowiczZ Gryczynski
Jul 19, 2005·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Gregor JungGraham R Fleming
Nov 2, 2001·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·P T SoC Y Dong
Dec 7, 2007·Optics Letters·G E Cragg, P T So
Sep 15, 2015·Applied Optics·Xuehua WangHanben Niu
Nov 2, 2019·Optics Express·Subir DasFu-Jen Kao
Nov 25, 2018·Optics Express·Michael KolanoGeorg Von Freymann
Mar 30, 1999·Analytical Biochemistry·H J LinJ R Lakowicz
Nov 24, 2004·Angewandte Chemie·Lars Kastrup, Stefan W Hell
Jun 22, 2021·Angewandte Chemie·Pavel Malý, Tobias Brixner
Feb 6, 2018·Analytical Chemistry·Andy Jing ChenJi-Xin Cheng
Aug 2, 2019·ACS Nano·Giulia ZaniniPaolo Bianchini
Jul 24, 2020·The Journal of Physical Chemistry. a·Libai HuangErik Grumstrup
Mar 26, 1999·Science·T NgP J Parker

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