BioTechniques (Jun 2007)

Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain

  • Le A. Trinh,
  • Marshall D. McCutchen,
  • Marianne Bonner-Fraser,
  • Scott E. Fraser,
  • Lloyd A. Bumm,
  • David W. McCauley

DOI
https://doi.org/10.2144/000112476
Journal volume & issue
Vol. 42, no. 6
pp. 756 – 759

Abstract

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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.