Evaluation and optimization of multiple fluorophore analysis of a Pseudomonas aeruginosa biofilm
Author(s)
Baird, Fiona J
Wadsworth, Marilyn P
Hill, Jane E
Griffith University Author(s)
Year published
2012
Metadata
Show full item recordAbstract
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.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.
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Journal Title
Journal of Microbiological Methods
Volume
90
Issue
3
Subject
Microbiology
Microbiology not elsewhere classified
Medical microbiology