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Published online 12 March 2007
Published in Soil Sci Soc Am J 71:620-631 (2007)
DOI: 10.2136/sssaj2006.0105
© 2007 Soil Science Society of America
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Fluorescent In Situ Hybridization and Micro-autoradiography Applied to Ecophysiology in Soil

Shane W. Rogersa,*, Thomas B. Moormanb and Say Kee Ongc

a U.S. Environmental Protection Agency, National Risk Management Research Lab., 26 W. Martin Luther King Dr., MS 421, Cincinnati, OH 45268
b USDA-ARS, National Soil Tilth Lab., 2150 Pammel Dr., Ames, IA 50011
c Dep. of Civil, Construction, and Environ. Eng., 486 Town Engineering Bldg., Iowa State Univ., Ames, IA 50011


Figure 1
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  		Fig. 1. Digital image processing of fluorescent in situ hybridization (FISH)–microautoradiography (MAR). Separate images for each color channel and the phase contrast counter image can be composited into a single image to simplify detection of cells that consume radioisotopic substrates. Stain DAPI (excitation = 385–400 nm, emission = 450–465 nm); probe EUB338-I, II, III (stain CY3; excitation = 530–560 nm, emission = 573–648 nm).

 

Figure 2
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  		Fig. 2. Autoradiogram (left side) and corresponding fluorescence image (right side) following 10-d autoradiographic film exposure to cells from 36-h aerobic incubations of polycyclic aromatic hydrocarbon contaminated aquifer sediments with [9-14C]phenanthrene. Silver grain clusters surrounding bacterial cells indicate active cellular incorporation of [9-14C]phenanthrene (A and B), which can be differentiated from bacteria that do not metabolize [9-14C]phenanthrene (C and D). Pink cells indicate Bacteria [EUB338(I, II, III) and blue cells are DAPI stained. Bar in autoradiogram = 10 µm. MAR = microautoradiography.

 




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