线粒体NAD(P)H荧光的时间分辨光谱及其在评估活细胞氧化状态中的应用。

Time-resolved spectrometry of mitochondrial NAD(P)H fluorescence and its applications for evaluating the oxidative state in living cells.

作者信息

Horilova Julia, Studier Hauke, Nadova Zuzana, Miskovsky Pavol, Chorvat Dusan, Chorvatova Alzbeta Marcek

机构信息

Department of Biophotonics, International Laser Center, Ilkovičova 3, 84104, Bratislava, Slovakia.

出版信息

Methods Mol Biol. 2015;1264:183-93. doi: 10.1007/978-1-4939-2257-4_17.

DOI:10.1007/978-1-4939-2257-4_17

PMID:25631014

Abstract

Time-resolved fluorescence spectrometry is a highly valuable technological tool to detect and characterize mitochondrial metabolic oxidative changes by means of endogenous fluorescence (Chorvat and Chorvatova, Laser Phys Lett 6: 175-193, 2009). Here, we describe the detection and measurement of endogenous mitochondrial NAD(P)H (nicotinamide adenine dinucleotide (phosphate)) fluorescence directly in living cultured cells using fluorescence lifetime spectrometry imaging after excitation with 405 nm picosecond (ps) laser. Time-correlated single photon counting (TCSPC) method is employed.

摘要

时间分辨荧光光谱法是一种极具价值的技术工具，可通过内源性荧光来检测和表征线粒体代谢氧化变化（Chorvat和Chorvatova，《激光物理通讯》6：175 - 193，2009年）。在此，我们描述了在活的培养细胞中，使用405纳米皮秒（ps）激光激发后通过荧光寿命光谱成像直接检测和测量内源性线粒体烟酰胺腺嘌呤二核苷酸（磷酸）（NAD(P)H）荧光的方法。采用了时间相关单光子计数（TCSPC）方法。