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Applied and Environmental Microbiology, April 2004, p. 2486-2493, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2486-2493.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Method for Collecting Air-Water Interface Microbes Suitable for Subsequent Microscopy and Molecular Analysis in both Research and Teaching Laboratories

Margaret C. Henk*

Socolofsky Microscopy Center, Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana

Received 14 May 2003/ Accepted 12 December 2003

A method has been developed for collecting air-water interface (AWI) microbes and biofilms that enables analysis of the same sample with various combinations of bright-field and fluorescence light microscopy optics, scanning and transmission electron microscopy (TEM), and atomic force microscopy. The identical sample is then subjected to molecular analysis. The sampling tool consists of a microscope slide supporting appropriate substrates, TEM grids, for example, that are removable for the desired protocols. The slide with its substrates is then coated with a collodion polymer membrane to which in situ AWI organisms adhere upon contact. This sampling device effectively separates the captured AWI bacterial community from the bulk water community immediately subtending. Preliminary data indicate that the AWI community differs significantly from the water column community from the same sample site when both are evaluated with microscopy and with 16S ribosomal DNA sequence-based culture-independent comparisons. This microbe collection method can be used at many levels in research and teaching.

* Corresponding author. Mailing address: Socolofsky Microscopy Center, Department of Biological Sciences, 202 Life Sciences Bldg., Louisiana State University, Baton Rouge, LA 70803. Phone: (225) 578-8860. Fax: (225) 578-2597. E-mail: henkmc{at}lsu.edu.

Applied and Environmental Microbiology, April 2004, p. 2486-2493, Vol. 70, No. 4
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.4.2486-2493.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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