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Standardization in microbial cytometry

CYTOMETRY of single bacteria still represents one of the most advanced measurement methods available to microbiologists, particularly when applied to statistically relevant numbers of single cells. However, progress in the field has been hampered by difficulties in signal measurement and interpretation of the data. With their small size microorganisms are at the edge of detection sensitivity of flow cytometers thus requiring extra care to avoid and minimize interference factors. Another problem comes from the physiological variability of microorganisms. The problem was discussed as part of the 2008 meeting on Microbial Analysis at the Single Cell Level (http://qbab.dbs. aber.ac.uk/sc2008) to get consensus and broader acceptance of microbial cytometry method and data standardization. Standardization can be applied in three domains, the sample manipulation, the measurement itself and the data interpretation as summarized in Figure 1. At the meeting in Bad Schandau, we started off an initiative to drive this forward. We will attempt to characterize a number of food grade/ pharmaceutical grade nonhazardous reference stains against a base panel of stains between a number of laboratories to achieve agreement over the staining methods and the respective patterns. Although it is expected that due to the nature of diversity some organisms will behave differently than the reference strains, this should facilitate identification of situations in which the problem does not come from the staining method or the measurement end but can be attributed to the diversity of the biology under investigation.