EMCCD-based spectrally resolved fluorescence correlation spectroscopy

被引：24

作者：

Bestvater, Felix ^{[1
,2
]}

Seghiri, Zahir ^{[1
]}

Kang, Moon Sik ^{[1
]}

Groener, Nadine ^{[3
]}

Lee, Ji Young ^{[1
]}

Im, Kang-Bin ^{[1
]}

Wachsmuth, Malte ^{[1
,3
]}

机构：

[1] Inst Pasteur Korea, Cell Biophys Grp, Songnam, Gyeonggi Do, South Korea

[2] German Canc Res Ctr, Light Microscopy Facil, D-69120 Heidelberg, Germany

[3] European Mol Biol Lab, Cell Biol & Biophys Unit, D-69117 Heidelberg, Germany

来源：

OPTICS EXPRESS | 2010年 / 18卷 / 23期

关键词：

CROSS-CORRELATION SPECTROSCOPY; QUANTUM DOTS; SINGLE MOLECULES; LIVE CELLS; CCD; FLUCTUATIONS; NANOCRYSTALS; CAMERA; PROBES;

D O I：

10.1364/OE.18.023818

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

We present an implementation of fluorescence correlation spectroscopy with spectrally resolved detection based on a combined commercial confocal laser scanning/fluorescence correlation spectroscopy microscope. We have replaced the conventional detection scheme by a prism-based spectrometer and an electron-multiplying charge-coupled device camera used to record the photons. This allows us to read out more than 80,000 full spectra per second with a signal-to-noise ratio and a quantum efficiency high enough to allow single photon counting. We can identify up to four spectrally different quantum dots in vitro and demonstrate that spectrally resolved detection can be used to characterize photophysical properties of fluorophores by measuring the spectral dependence of quantum dot fluorescence emission intermittence. Moreover, we can confirm intracellular cross-correlation results as acquired with a conventional setup and show that spectral flexibility can help to optimize the choice of the detection windows. (C) 2010 Optical Society of America

引用

页码：23818 / 23828

页数：11

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