注意：使用互补金属氧化物半导体-单光子雪崩二极管阵列在活细胞中进行多共焦荧光相关光谱分析。

Note: multi-confocal fluorescence correlation spectroscopy in living cells using a complementary metal oxide semiconductor-single photon avalanche diode array.

作者信息

Kloster-Landsberg M, Tyndall D, Wang I, Walker R, Richardson J, Henderson R, Delon A

机构信息

University of Grenoble 1/CNRS, LIPhy UMR 5588, F-38041 Grenoble, France.

出版信息

Rev Sci Instrum. 2013 Jul;84(7):076105. doi: 10.1063/1.4816156.

DOI:10.1063/1.4816156

PMID:23902122

Abstract

Living cells are heterogeneous and rapidly changing biological samples. It is thus desirable to measure molecular concentration and dynamics in many locations at the same time. In this note, we present a multi-confocal setup capable of performing simultaneous fluorescence correlation spectroscopy measurements, by focusing the spots with a spatial light modulator and acquiring data with a monolithic 32 × 32 single-photon avalanche photodiode array. A post-processing method is proposed to correct cross-talk effects between neighboring spots. We demonstrate the applicability of our system by simultaneously measuring the diffusion of free enhanced Green Fluorescent Protein (eGFP) molecules at nine different points in living cells.

摘要

活细胞是异质且快速变化的生物样本。因此，期望能同时在多个位置测量分子浓度和动力学。在本报告中，我们展示了一种多共聚焦装置，该装置能够通过使用空间光调制器聚焦光斑并利用单片32×32单光子雪崩光电二极管阵列采集数据来进行同步荧光相关光谱测量。我们提出了一种后处理方法来校正相邻光斑之间的串扰效应。我们通过同时测量活细胞中九个不同点处的游离增强型绿色荧光蛋白（eGFP）分子的扩散，证明了我们系统的适用性。