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Fluorescence in situ hybridization of freshwater fungi
Citation key Baschien2001a
Author Baschien, C. and Manz, W. and Neu, T. R. and Szewzyk, U.
Pages 371–381
Year 2001
DOI 10.1002/1522-2632(200107)86:4/5<371::AID-IROH371>3.0.CO;2-P
Journal International Review Of Hydrobiology
Volume 86
Number 4-5
Publisher Wiley-V C H Verlag Gmbh
Abstract Based on computer assisted comparative sequence analysis, the 18S rRNA targeted fungal oligonucleotide probe FUN1429 was designed and evaluated for the identification of in situ metabolically active Eumycota. The general accessibility of fungal cells for fluorescent oligonucleotides was tested by whole cell hybridizations. Additionally, the influences of different growth media, age of the fungal specimen and the composition of permeability buffers were assessed. All strains showed clear fluorescence hybridization signals after visualization with confocal laser scanning microscopy (CSLM). In contrast fluorescence hybridization signals were hardly detectable by conventional epifluorescence microscopy due to strong fungal autofluorescence. The inherent autofluorescence emitted from the strains increased with the age of cultures, but was significantly decreased by chitinase treatment prior to in situ hybridization. The composition of the growth media showed no measurable effect on signal intensity.
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