Bacterial Suspensions Deposited on Microbiological Filter Material for Rapid Laser-Induced Breakdown Spectroscopy Identification

Applied Spectroscopy
Dylan J MalenfantSteven J Rehse

Abstract

Four species of bacteria, E. coli, S. epidermidis, M. smegmatis, and P. aeruginosa, were harvested from agar nutrient medium growth plates and suspended in water to create liquid specimens for the testing of a new mounting protocol. Aliquots of 30 µL were deposited on standard nitrocellulose filter paper with a mean 0.45 µm pore size to create highly flat and uniform bacterial pads. The introduction of a laser-based lens-to-sample distance measuring device and a pair of matched off-axis parabolic reflectors for light collection improved both spectral reproducibility and the signal-to-noise ratio of optical emission spectra acquired from the bacterial pads by laser-induced breakdown spectroscopy. A discriminant function analysis and a partial least squares-discriminant analysis both showed improved sensitivity and specificity compared to previous mounting techniques. The behavior of the spectra as a function of suspension concentration and filter coverage was investigated, as was the effect on chemometric cell classification of sterilization via autoclaving.

References

Nov 5, 2003·Applied Optics·Stéphane MorelJacques Amouroux
Dec 3, 2003·Applied Spectroscopy·John D HyblSteven G Buckley
Feb 19, 2011·Analytical and Bioanalytical Chemistry·Jennifer L Gottfried
Mar 23, 2011·Analytical and Bioanalytical Chemistry·Cleon BarnettShree Singh
Aug 10, 2011·Analytical and Bioanalytical Chemistry·Dawn E LewisMaurice D Edington
Aug 15, 2013·Journal of Agricultural and Food Chemistry·Rosalie A MultariJohn E Gustafson

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Methods Mentioned

BETA
scanning electron microscopy

Software Mentioned

DA
DFA
ESAWIN
PLS

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