Evaluation and optimization of multiple fluorophore analysis of a Pseudomonas aeruginosa biofilm

被引:25
作者
Baird, Fiona J. [1 ]
Wadsworth, Marilyn P. [2 ]
Hill, Jane E. [1 ]
机构
[1] Univ Vermont, Sch Engn, Burlington, VT 05405 USA
[2] Univ Vermont, Coll Med, Microscopy Imaging Ctr, Burlington, VT USA
关键词
Biofilm; Spectral imaging; Pseudomonas aeruginosa; EXTRACELLULAR POLYMERIC SUBSTANCES; FLUORESCENCE MICROSCOPY; CONFOCAL MICROSCOPY; MOUNTING MEDIA; CELLS;
D O I
10.1016/j.mimet.2012.05.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Conventional laser scanning microscopy for multiple fluorescent stains can be a useful tool if the problems of autofluorescence and cross-talk are eliminated. The technique of spectral imaging was employed to unmix five different fluorophores - ranging in emission from 435 to 665 nm - applied to a Pseudomonas aeruginosa biofilm with overlapping spectra and which was not possible using traditional channel mode operation. Using lambda scanning and linear unmixing, the five fluorophores could be distinguished with regions of differentiation apparent. (C) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:192 / 196
页数:5
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