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An improved method for collecting and staining microorganisms for enumeration by fluorescence light microscopy.
F A Williamson1, K R Palframan
1Shell Research Limited, Sittingbourne Research Centre, Kent, U.K.
Journal of Microscopy
|June 1, 1989
Summary
A new Anopore membrane filter improves microbial enumeration. This method enhances image contrast for accurate total and viable microorganism counts using automated image analysis.
Area of Science:
- Microbiology
- Analytical Chemistry
- Biotechnology
Background:
- Previous methods for microbial enumeration using fluorescence microscopy on membrane filters were suboptimal.
- Limitations included non-specific filter staining and filter surface irregularities hindering automated image analysis.
Purpose of the Study:
- To evaluate the Anopore membrane filter for improved enumeration of total and viable microorganisms.
- To develop a robust and rapid method suitable for automated microbial population analysis.
Main Methods:
- Utilized Anopore membrane filters for bacterial staining with ethidium bromide.
- Employed fluorescence light microscopy and automated image analysis techniques.
- Assessed filter properties including inorganic construction, porosity, low adsorption, and hydrophilicity.
Main Results:
- Anopore membranes produced high-contrast images of stained bacteria.
- Images were suitable for automated image analysis using simple grey-level thresholding.
- The method demonstrated potential for a fully automated microbial enumeration procedure.
Conclusions:
- Anopore membrane filters overcome limitations of previous filter types for microbial enumeration.
- The developed method provides a basis for rapid, robust, and automated counting of microbial populations.