title: Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain.
abstract: 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.
authors.affiliation: Beckman Institute, California Institute of Technology, Pasadena, CA 91125, USA. ltrinh@caltech.edu
authors.forename: Le A
authors.initials: le A
authors.lastname: Trinh
authors.forename: Marshall D
authors.initials: MD
authors.lastname: McCutchen
authors.forename: Marianne
authors.initials: M
authors.lastname: Bonner-Fraser
authors.forename: Scott E
authors.initials: SE
authors.lastname: Fraser
authors.forename: Lloyd A
authors.initials: LA
authors.lastname: Bumm
authors.forename: David W
authors.initials: DW
authors.lastname: McCauley
mesh_terms: D000818:Animals; D024722:Cardiac Myosins / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007211:Indoles / Q000737:chemistry*; D007798:Lampreys / Q000196:embryology; D018613:Microscopy, Confocal; D009580:Nitroblue Tetrazolium / Q000737:chemistry*; D013194:Staining and Labeling; D015027:Zebrafish / Q000196:embryology
chemical_list: D007211:Indoles; D009580:Nitroblue Tetrazolium; C035455:5-bromo-4-chloro-3-indoxyl phosphate; D024722:Cardiac Myosins