利用三维荧光寿命成像技术对活组织中的钙进行定量成像。

Quantitative Imaging of Ca by 3D-FLIM in Live Tissues.

作者信息

Rakymzhan Asylkhan, Radbruch Helena, Niesner Raluca A

机构信息

Deutsches Rheuma-Forschungszentrum, a Leibniz Institute, Charitéplatz 1, 10117, Berlin, Germany.

Neuropathology, Charité-Universitätsmedizin, Charitéplatz 1, 10117, Berlin, Germany.

出版信息

Adv Exp Med Biol. 2017;1035:135-141. doi: 10.1007/978-3-319-67358-5_9.

DOI:10.1007/978-3-319-67358-5_9

PMID:29080135

Abstract

The calcium concentration within living cells is highly dynamic and, for many cell types, a reliable indicator of the functional state of the cells-both of isolated cells, but even, more important, of cells in tissue. In order to dynamically quantify intracellular calcium levels, various genetically encoded calcium sensors have been developed-the best of which are those based on Förster resonant energy transfer (FRET). Here we present a fluorescence lifetime imaging (FLIM) method to measure FRET in such a calcium sensor (TN L15) in neurons of hippocampal slices and of the brain stem of anesthetized mice. The method gives the unique opportunity to determine absolute neuronal calcium concentrations in the living organism.

摘要

活细胞内的钙浓度具有高度动态性，对于许多细胞类型而言，它是细胞功能状态的可靠指标——不仅适用于分离的细胞，更重要的是，适用于组织中的细胞。为了动态量化细胞内钙水平，人们开发了各种基因编码的钙传感器，其中最好的是基于Förster共振能量转移（FRET）的传感器。在此，我们展示了一种荧光寿命成像（FLIM）方法，用于测量麻醉小鼠海马切片和脑干神经元中此类钙传感器（TN L15）中的FRET。该方法为确定活生物体中的绝对神经元钙浓度提供了独特的机会。