Microorganisms detection Fluorescence In Situ Hybridization Fluorophores
Sociedade Portuguesa de Microbiologia; Sociedade Portuguesa de Biotecnología; Católica Escola Superior de Biotecnologia, Porto;,
FISH has been applied in many ecological and phylogenetic studies becoming the method of choice for the direct detection and
identification of microorganisms in their natural environments . For reliable results, it is crucial to minimise or avoid
background fluorescence and cellular autofluorescence but also to maximise the specific FISH signals obtained. Red-emitting
dyes have been usually used for eliminating the problems of non-specific fluorescence interference. Therefore, in this work we
evaluated the role of three of these fluorophores (Alexa Fluor 647 (AF647), ATTO 647N and Cy5) on the reliability of the RNAFISH
results obtained both with a universal and a specie-specific probe.