Fluorescence lifetime images and correlation spectra obtained by multidimensional time-correlated single photon counting

被引：52

作者：

Becker, W

Bergmann, A

Haustein, E

Petrasek, Z

Schwille, P

Biskup, C

Kelbauskas, L

Benndorf, K

Klöcker, N

Anhut, T

Riemann, I

König, K

机构：

[1] Becker & Hickel GMBH, D-12277 Berlin, Germany

[2] Biotec TU Dresden, Inst Biophys, D-01307 Dresden, Germany

[3] Univ Jena, Jena Inst Physiol 2, D-07740 Jena, Germany

[4] Univ Freiburg, Dept Physiol 2, D-79104 Freiburg, Germany

[5] Fraunhofer Inst Biomed Engn, D-66386 St Ingbert, Germany

来源：

MICROSCOPY RESEARCH AND TECHNIQUE | 2006年 / 69卷 / 03期

关键词：

time-correlated single photon counting (TCSPC); fluorescence lifetime imaging (FLIM); Forster resonance energy transfer (FRET); autofluorescence; fluorescence correlation spectroscopy (FCS);

D O I：

10.1002/jemt.20251

中图分类号：

R602 [外科病理学、解剖学]; R32 [人体形态学];

学科分类号：

100101 ;

摘要：

Multidimensional time-correlated single photon counting (TCSPC) is based on the excitation of the sample by a high-repetition rate laser and the detection of single photons of the fluorescence signal in several detection channels. Each photon is characterized by its arrival time in the laser period, its detection channel number, and several additional variables such as the coordinates of an image area, or the time from the start of the experiment. Combined with a confocal or two-photon laser scanning microscope and a pulsed laser, multidimensional TCSPC makes a fluorescence lifetime technique with multiwavelength capability, near-ideal counting efficiency, and the capability to resolve multiexponential decay functions. We show that the same technique and the same hardware can be used for precision fluorescence decay analysis and fluorescence correlation spectroscopy (FCS) in selected spots of a sample.

引用

页码：186 / 195

页数：10

共 70 条

[1] Fluorescence resonance energy transfer determinations using multiphoton fluorescence lifetime imaging microscopy to characterize amyloid-beta plaques [J].