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Published June 2007 | public
Journal Article

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.

Additional Information

© 2007 BioTechniques. Received 5 March 2007; accepted 6 April 2007. L.A.T., M.D.M., L.A.B., and D.W.M. contributed equally to this work. M.D.M. thanks the National Science Foundation (NSF) Louis Stokes Oklahoma Alliance for Minority Participation, Bridges to the Doctorate Program for a graduate research fellowship. This work was supported by the NSF CAREER grant no. CHE-0239803, the Center for Physics in Nanostructures, NSF MRSEC no. DMR-0080054, Oklahoma EPSCoR, and NIH grants no. R01 HL078694 and P01 HD037105. The authors declare no competing interests.

Additional details

Created:
August 19, 2023
Modified:
October 17, 2023