Real-time Analysis of Metabolic Activity Within Lactobacillus acidophilus by Phasor Fluorescence Lifetime Imaging Microscopy of NADH

被引：20

作者：

Torno, Keenan ^{[1
]}

Wright, Belinda K. ^{[1
]}

Jones, Mark R. ^{[1
]}

Digman, Michelle A. ^{[2
]}

Gratton, Enrico ^{[2
]}

Phillips, Michael ^{[1
]}

机构：

[1] Univ Western Sydney, Sch Sci & Hlth, Richmond, NSW, Australia

[2] Univ Calif Irvine, Fluorescence Dynam Lab, Dept Biomed Engn, Irvine, CA USA

来源：

CURRENT MICROBIOLOGY | 2013年 / 66卷 / 04期

关键词：

CELLS;

D O I：

10.1007/s00284-012-0285-2

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Nicotinamide adenine dinucleotide (NADH) is an endogenous fluorescent molecule commonly used as a metabolic biomarker. Fluorescence lifetime imaging microscopy (FLIM) is a method in which the fluorescence decay is measured at each pixel of an image. While the fluorescence spectrum of free and protein-bound NADH is very similar, free and protein-bound NADH display very different decay profiles. Therefore, FLIM can provide a way to distinguish free/bound NADH at the level of single bacteria within biological samples. The phasor technique is a graphical method to analyse the entire image and to produce a histogram of pixels with different decay profile. In this study, NADH fluorescence decay profiles within Lactobacillus acidophilus samples treated using different protocols indicated discernible variations. Clear distinctions between fluorescence decay profiles of NADH in samples of artificially heightened metabolic activity in comparison to those of samples lacking an accessible carbon source were obtained.

引用

页码：365 / 367

页数：3

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