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Applied and Environmental Microbiology, January 2003, p. 641-643, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.641-643.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Evaluation of the Redox Dye 5-Cyano-2,3-Tolyl-Tetrazolium Chloride for Activity Studies by Simultaneous Use of Microautoradiography and Fluorescence In Situ Hybridization

Jeppe Lund Nielsen, Marilena Aquino de Muro,{dagger} and Per Halkjær Nielsen*

Department of Environmental Engineering, Aalborg University, DK-9000 Aalborg, Denmark

Received 22 April 2002/ Accepted 16 July 2002

Three microscopic in situ techniques were used simultaneously to investigate viability and activity on a single-cell level in activated sludge. The redox dye 5-cyano-2,3-tolyl-tetrazolium chloride (CTC) was compared with microautoradiography (MAR) and fluorescence in situ hybridization (FISH) to indicate activity of cells in Thiothrix filaments and in single floc-forming bacteria. The signals from MAR and FISH correlated well, whereas only 65% of the active Thiothrix cells and 41% of all single cells were detectable by CTC reduction, which mainly targeted the most active cells.

* Corresponding author. Mailing address: Department of Environmental Engineering, Aalborg University, Sohngaardsholmsvej 57, DK-9000 Aalborg, Denmark, Phone: 45 96358080. Fax: 45 98 14 25 55. E-mail: phn{at}bio.auc.dk.

{dagger} Present address: CABI Bioscience UK Centre [Egham], Biosystematics and Molecular Biology, Egham, Surrey TW20 9TY, United Kingdom.

Applied and Environmental Microbiology, January 2003, p. 641-643, Vol. 69, No. 1
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.1.641-643.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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