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Applied and Environmental Microbiology, February 2009, p. 758-771, Vol. 75, No. 3
0099-2240/09/$08.00+0     doi:10.1128/AEM.00819-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Modular Spectral Imaging System for Discrimination of Pigments in Cells and Microbial Communities{triangledown} ,{dagger}

Lubos Polerecky,1* Andrew Bissett,1 Mohammad Al-Najjar,1 Paul Faerber,1 Harald Osmers,1 Peter A. Suci,2 Paul Stoodley,3 and Dirk de Beer1

Max Planck Institute for Marine Microbiology, Celsiusstrasse 1, 28359 Bremen, Germany,1 Center for Biofilm Engineering, 366 EPS—P.O. Box 173980, Montana State University—Bozeman, Bozeman, Montana 59717-3980,2 Center for Genomic Sciences, Allegheny-Singer Research Institute, 1107 11th Floor South Tower, 320 East North Avenue, Pittsburgh, Pennsylvania 15212-47723

Received 10 April 2008/ Accepted 24 November 2008

Here we describe a spectral imaging system for minimally invasive identification, localization, and relative quantification of pigments in cells and microbial communities. The modularity of the system allows pigment detection on spatial scales ranging from the single-cell level to regions whose areas are several tens of square centimeters. For pigment identification in vivo absorption and/or autofluorescence spectra are used as the analytical signals. Along with the hardware, which is easy to transport and simple to assemble and allows rapid measurement, we describe newly developed software that allows highly sensitive and pigment-specific analyses of the hyperspectral data. We also propose and describe a number of applications of the system for microbial ecology, including identification of pigments in living cells and high-spatial-resolution imaging of pigments and the associated phototrophic groups in complex microbial communities, such as photosynthetic endolithic biofilms, microbial mats, and intertidal sediments. This system provides new possibilities for studying the role of spatial organization of microorganisms in the ecological functioning of complex benthic microbial communities or for noninvasively monitoring changes in the spatial organization and/or composition of a microbial community in response to changing environmental factors.

* Corresponding author. Mailing address: Max Planck Institute for Marine Microbiology, Celsiusstrasse 1, 28359 Bremen, Germany. Phone: 49 (0) 421 2028 834. Fax: 49 (0) 421 2028 690. E-mail: lpolerec{at}mpi-bremen.de

{triangledown} Published ahead of print on 12 December 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, February 2009, p. 758-771, Vol. 75, No. 3
0099-2240/09/$08.00+0     doi:10.1128/AEM.00819-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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