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Applied and Environmental Microbiology, July 2007, p. 4508-4514, Vol. 73, No. 14
0099-2240/07/$08.00+0     doi:10.1128/AEM.00733-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Suitability of Flow Cytometry for Estimating Bacterial Biovolume in Natural Plankton Samples: Comparison with Microscopy Data{triangledown}

Marisol Felip,1* Stefan Andreatta,2 Ruben Sommaruga,3 Viera Straskrábová,4 and Jordi Catalan5

Unitat de Limnologia (CSIC-UB), Departament d'Ecologia, Universitat de Barcelona, E-08028 Barcelona, Spain,1 Synedra Information Technologies, A-6020 Innsbruck, Austria,2 Laboratory of Aquatic Photobiology and Plankton Ecology, Institute of Ecology, University of Innsbruck, A-6020 Innsbruck, Austria,3 Czech Academy of Sciences, CZ-37005 Ceské Budejovice, Czech Republic,4 Unitat de Limnologia (CSIC-UB), Centre d'Estudis Avançats de Blanes, CSIC, E-17300 Blanes, Spain5

Received 2 April 2007/ Accepted 11 May 2007

The relationship between flow cytometry data and epifluorescence microscopy measurements was assessed in bacterioplankton samples from 80 lakes to estimate bacterial biovolume and cell size distribution. The total counts of 4',6'-diamidino-2-phenylindole-stained cells estimated by both methods were significantly related, and the slope of their linear regression was not significantly different from 1, indicating that both methods produce very similar estimates of bacterial abundance. The relationships between side scatter (SSC) and 4',6'-diamidino-2-phenylindole fluorescence and cell volume (microscopy values) were improved by binning of the data in three frequency classes for each, but further increases in the number of classes did not improve these relationships. Side scatter was the best cell volume predictor, and significant relationships were observed between the SSC classes and the smallest (R2 = 0.545, P < 0.001, n = 80) and the largest (R2 = 0.544, P < 0.001, n = 80) microscopy bacterial-size classes. Based on these relationships, a reliable bacterial biomass estimation was obtained from the SSC frequency classes. Our study indicates that flow cytometry can be used to properly estimate bacterioplankton biovolume, with an accuracy similar to those of more time-consuming microscopy methods.

* Corresponding author. Mailing address: Unitat de Limnologia (CSIC-UB), Departament d'Ecologia, Universitat de Barcelona, Avgd Diagonal 645, 08028 Barcelona, Spain. Phone: 34 934021507. Fax: 34 4111438. E-mail: mfelip{at}ub.edu

{triangledown} Published ahead of print on 18 May 2007.

Applied and Environmental Microbiology, July 2007, p. 4508-4514, Vol. 73, No. 14
0099-2240/07/$08.00+0     doi:10.1128/AEM.00733-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.





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