Roles of Triton X-100 in NADPH-diaphorase histochemistry.
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.
Fang
S
S
Department of Internal Medicine, University of Iowa College of Medicine, Iowa City.
Christensen
J
J
Conklin
J L
JL
Murray
J A
JA
Clark
G
G
D000818:Animals; D001391:Azo Compounds; D003106:Colon / Q000737:chemistry / Q000201:enzymology; D004947:Esophagus / Q000737:chemistry / Q000201:enzymology; D005260:Female; D006651:Histocytochemistry / Q000379:methods*; D007421:Intestine, Small / Q000737:chemistry / Q000201:enzymology; D008297:Male; D009252:NADPH Dehydrogenase / Q000032:analysis*; D017830:Octoxynol; D009893:Opossums
D001391:Azo Compounds; C033302:diformazan dye; D017830:Octoxynol; D009252:NADPH Dehydrogenase