Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain.
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.
Trinh
Le A
le A
Beckman Institute, California Institute of Technology, Pasadena, CA 91125, USA. ltrinh@caltech.edu
McCutchen
Marshall D
MD
Bonner-Fraser
Marianne
M
Fraser
Scott E
SE
Bumm
Lloyd A
LA
McCauley
David W
DW
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
D007211:Indoles; D009580:Nitroblue Tetrazolium; C035455:5-bromo-4-chloro-3-indoxyl phosphate; D024722:Cardiac Myosins