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Applied and Environmental Microbiology, August 2005, p. 4801-4808, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4801-4808.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Optically Transparent Porous Medium for Nondestructive Studies of Microbial Biofilm Architecture and Transport Dynamics

Andrew P. Leis,1* Sven Schlicher,2 Hilmar Franke,2 and Martin Strathmann1

University of Duisburg-Essen, Institute for Interface Biotechnology, Department of Aquatic Microbiology, Geibelstr. 41, 47057 Duisburg, Germany,1 University of Duisburg-Essen, Institute for Physics, Lotharstr. 65, 47057 Duisburg, Germany2

Received 20 December 2004/ Accepted 3 March 2005

We describe a novel and noninvasive, microscopy-based method for visualizing the structure and dynamics of microbial biofilms, individual fluorescent microbial cells, and inorganic colloids within a model porous medium. Biofilms growing in flow cells packed with granules of an amorphous fluoropolymer could be visualized as a consequence of refractive index matching between the solid fluoropolymer grains and the aqueous immersion medium. In conjunction with the capabilities of confocal microscopy for nondestructive optical sectioning, the use of amorphous fluoropolymers as a solid matrix permits observation of organisms and dynamic processes to a depth of 2 to 3 mm, whereas sediment biofilms growing in sand-filled flow cells can only be visualized in the region adjacent to the flow cell wall. This method differs fundamentally from other refractive index-matching applications in that optical transparency was achieved by matching a solid phase to water (and not vice versa), thereby permitting real-time microscopic studies of particulate-containing, low-refractive-index media such as biological and chromatographic systems.

* Corresponding author. Present address: Max Planck Institute for Biochemistry, Department of Molecular Structural Biology, Am Klopferspitz 18, D-82152 Martinsried, Germany. Phone: 49 (89) 8578 2033. Fax: 49 (89) 8578 2641. E-mail: leis{at}biochem.mpg.de.

Applied and Environmental Microbiology, August 2005, p. 4801-4808, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4801-4808.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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