Abstract
Bacteria were either heat fixed on microscope slides or filtered with 0.2 micron-pore-size Nuclepore filters. The samples were stained with 4',6-diamidino-2-phenylindole (DAPI) for total staining and with polyvalent rabbit antibodies and fluorescein isothiocyanate-conjugated swine anti-rabbit antibodies for specific staining. By switching between two different optical filter packages in the microscope, only one sample was needed for determining both total and specific counts of bacteria. False-positive counts and other artifacts that occur with antibody staining were easily distinguished when individual fluorescent particles were checked for DAPI fluorescence. The method for applying the general stain to membrane filters was performed quickly and simply by using a DAPI-soaked polypropylene filter that lay beneath the Nuclepore filter which collected the sample.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bergström I., Heinänen A., Salonen K. Comparison of acridine orange, acriflavine, and bisbenzimide stains for enumeration of bacteria in clear and humic waters. Appl Environ Microbiol. 1986 Mar;51(3):664–667. doi: 10.1128/aem.51.3.664-667.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hobbie J. E., Daley R. J., Jasper S. Use of nuclepore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol. 1977 May;33(5):1225–1228. doi: 10.1128/aem.33.5.1225-1228.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howgrave-Graham Alan R., Steyn Pieter L. Application of the Fluorescent-Antibody Technique for the Detection of Sphaerotilus natans in Activated Sludge. Appl Environ Microbiol. 1988 Mar;54(3):799–802. doi: 10.1128/aem.54.3.799-802.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Muyzer G., de Bruyn A. C., Schmedding D. J., Bos P., Westbroek P., Kuenen G. J. A Combined Immunofluorescence-DNA-Fluorescence Staining Technique for Enumeration of Thiobacillus ferrooxidans in a Population of Acidophilic Bacteria. Appl Environ Microbiol. 1987 Apr;53(4):660–664. doi: 10.1128/aem.53.4.660-664.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- doi: 10.1177/003591574203600109. [DOI] [PMC free article] [Google Scholar]
- Paul J. H. Use of hoechst dyes 33258 and 33342 for enumeration of attached and planktonic bacteria. Appl Environ Microbiol. 1982 Apr;43(4):939–944. doi: 10.1128/aem.43.4.939-944.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rennie R. J., Schmidt E. L. Immunofluorescence studies of Nitrobacter populations in soils. Can J Microbiol. 1977 Aug;23(8):1011–1017. doi: 10.1139/m77-150. [DOI] [PubMed] [Google Scholar]
- Stahl M. L., Williams F. D. Immunofluorescent evidence of Proteus mirabilis swarm cell formation on sterilized rat feces. Appl Environ Microbiol. 1981 Mar;41(3):801–806. doi: 10.1128/aem.41.3.801-806.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]