Detection and Quantification of Viable Airborne Bacteria and Fungi Using Solid-phase Cytometry

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2009 Jan 31

PMID 19180091

Citations 2

Authors

Lies M E Vanhee

Hans J Nelis

Tom Coenye

Affiliations

Soon will be listed here.

Abstract

This protocol describes the use of solid-phase cytometry for the enumeration of airborne bacteria and fungi. In contrast with conventional methods, accurate results can be obtained in real time, especially for air samples with low numbers of microorganisms. Air samples are collected by impaction on a water-soluble polymer that is subsequently dissolved. Part of the sample can be filtered over two membrane filters with different pore sizes. One filter is used to obtain a total count of all viable microorganisms, and a second filter is used to determine the number of airborne fungi. Microorganisms present on the filter are labeled with a viability substrate and subsequently detected and quantified using a solid-phase cytometer. The detected spots are microscopically validated using an epifluorescence microscope to discriminate between bacteria, fungi and fluorescent particles. The whole procedure takes 5 h to complete and results in the accurate quantification of airborne bacteria and fungi for samples with a low or high microbial load.

Citing Articles

Rapid detection of total and viable Legionella pneumophila in tap water by immunomagnetic separation, double fluorescent staining and flow cytometry.

Keserue H, Baumgartner A, Felleisen R, Egli T Microb Biotechnol. 2012; 5(6):753-63.

PMID: 23062200 PMC: 3815896. DOI: 10.1111/j.1751-7915.2012.00366.x.

Rapid and direct quantification of viable Candida species in whole blood by use of immunomagnetic separation and solid-phase cytometry.

Vanhee L, Meersseman W, Lagrou K, Maertens J, Nelis H, Coenye T J Clin Microbiol. 2010; 48(4):1126-31.

PMID: 20129966 PMC: 2849573. DOI: 10.1128/JCM.00035-10.

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