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Fluorescent in situ hybridization heating pretreatment: The key is temperature control
Author(s) -
Tojo Marta,
Couso Elena,
VázquezBoquete Angel,
PérezBecerra Raquel,
GarcíaCaballero Tomás,
Forteza Jerónimo,
Fraga Máximo
Publication year - 2010
Publication title -
pathology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.73
H-Index - 74
eISSN - 1440-1827
pISSN - 1320-5463
DOI - 10.1111/j.1440-1827.2010.02600.x
Subject(s) - fish <actinopterygii> , in situ , in situ hybridization , temperature control , fluorescence , digestion (alchemy) , buffer (optical fiber) , fluorescence in situ hybridization , chemistry , materials science , chromatography , biology , computer science , biochemistry , mechanical engineering , messenger rna , engineering , fishery , telecommunications , physics , organic chemistry , quantum mechanics , gene , chromosome
Fluorescent in situ hybridization (FISH) is a very useful tool for diagnostic and prognostic purposes in pathology. However, many laboratories still experience troubles when applying FISH to paraffin material. To overcome these difficulties, different pretreatments which include enzymatic digestion have been described. Usually, previous to digestion, a heating step is performed. The aim of this study was to compare the efficiency of the heating step with different buffers and different heating methods. We conclude that the main factor in the heating pretreatment is the temperature control, irrespective of the buffer used. Best results are obtained with any buffer by heating the slides to 99°C for 15 min followed by 10 min at room temperature.