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J Microbiol Methods. 2000 Sep;42(1):29-38.

Single-cell analysis of bacteria by Raman microscopy: spectral information on the chemical composition of cells and on the heterogeneity in a culture.

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Institute of Chemical Engineering, Fuel and Environmental Technology, Vienna University of Technology, Getreidemarkt 9/159, 1060, Vienna, Austria.


In the acetone-butanol (ABE) fermentation process, the utilised organisms from the group of the solventogenic Clostridia go through a complex cell-cycle. The role of different cell types in product formation is not understood in detail yet. We aim to use Raman spectroscopy to characterise the population distribution in Clostridium cultures. Cell suspensions were dried on calcium fluoride carriers. Raman spectra of single cells were obtained using a confocal Raman microscope (Dilor, Lille, France). The laser beam was focused on individual cells through the microscope objective. Spectra with good signal-to-noise ratio were obtained. Cells of different morphology, but also apparently similar cells, showed different spectra. Several cell components could be detected and varied in quantity. Compared to other methods for single-cell analysis, the new method is much more time-consuming to analyse one individual cell. However, a large amount of chemical information is obtained from each single cell in a non-destructive, non-invasive way. Raman microscopy appears to be a suitable method for studying population distributions in bacterial cultures.

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