Open AccessFluorescent in Situ Hybridization Employing the Conventional NBT/BCIP Chromogenic Stain
Author(s) -
Le A. Trinh,
Marshall D. McCutchen,
Marianne Bonner-Fraser,
Scott E. Fraser,
Lloyd A. Bumm,
David W. McCauley
Publication year - 2007
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000112476
Subject(s) - chromogenic , stain , fluorescence , in situ , microbiology and biotechnology , staining , chemistry , in situ hybridization , confocal microscopy , confocal , fluorescence microscope , chromogenic in situ hybridization , microscopy , gene expression , biology , gene , optics , biochemistry , chromatography , genetics , physics , organic chemistry
In situ hybridization techniques typically employ chromogenic staining by enzymatic amplification to detect domains of gene expression. We demonstrate the previously unreported near infrared (NIR) fluorescence of the dark purple stain formed from the commonly used chromogens, nitro blue tetrazolium (NBT) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP). The solid reaction product has significant fluorescence that enables the use of confocal microscopy to generate high-resolution three-dimensional (3-D) imaging of gene expression.
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