Roles of Triton X-100 in NADPH-diaphorase histochemistry

S. Fang, J. Christensen, J. L. Conklin, J. A. Murray, G. Clark

Research output: Contribution to journalArticlepeer-review

18 Scopus citations


Triton X-100 is widely but not universally used in NADPH-diaphorase histochemical staining. We investigated its effect on the staining and examined nitroblue diformazan (NBF) production under the influence of Triton X-100. Exposure of opossum esophagus, intestine, and colon tissues to Triton X-100 before staining enhanced staining of nerve cells and fibers and suppressed staining of non-neural structures. Long exposures and high concentrations nearly abolished the staining of non-neural structures and decreased the staining of nerves. The use of an incubation medium containing Triton X-100 achieved the best staining of nerve cells and fibers. Addition of Triton X-100 to the incubation medium changed its color from yellow to purple; in the presence of tissues, this color change occurred much more quickly. Spectral analysis showed that Triton X-100 increases the rate of NBF formation in the presence of tissue supernatant. Triton X-100 increases it less in the absence of tissue supernatant. Therefore, Triton X-100 improves the histochemical staining, probably by catalyzing the activity of NADPH- diaphorase, by keeping the extracellular NBF in solution and thus suppressing the staining of non-neural structures, and by increasing the permeability of cell membranes.

Original languageEnglish (US)
Pages (from-to)1519-1524
Number of pages6
JournalJournal of Histochemistry and Cytochemistry
Issue number11
StatePublished - 1994


  • Colon
  • Enzyme histochemistry
  • Esophagus
  • Intestine
  • NADPH-diaphorase histochemistry
  • NBT
  • Nitric oxide synthase
  • Opossum
  • Triton X-100
  • β- NADPH

ASJC Scopus subject areas

  • Anatomy
  • Histology


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