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Tetramethyl benzidine for horseradish peroxidase neurohistochemistry: a non-carcinogenic blue reaction product with superior sensitivity for visualizing neural afferents and efferents

MM Mesulam

Tetramethyl benzidine (TMB) is a presumptively non-carcinogenic chromogen which yields a blue reaction-product at sites of horseradish peroxidase activity. Sixty-six distinct procedures were performed in rats and monkeys in order to determine the optimal incubation parameters for TMB. As a result, a procedure is recommended whose sensitivity greatly surpasses that of a previously described benzidine dihydrochloride method. Indeed, the sensitivity of this new method in demonstrating retrograde transport is markedly superior to that of the previously described benzidine dihydrochloride method. Furthermore, as a consequence of this enhanced sensitivity, many efferent connections of the injection site are also visualized. The injection site demonstrated by this TMB procedure is significantly larger than the one demonstrated when benzidine dihydrochloride or diaminobenzidine is used as a chromogen. Finally, this TMB procedure has been compared to two other TMB procedures and found to provide superior morphology and sensitivity.

Volume 26, Issue 2, pp. 106-117, 02/01/1978
Copyright © 1978 by The Histochemical Society


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Science, August 6, 1982; 217(4559): 553 - 555.
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 Arch Otolaryngol Head Neck SurgHome page
S. M. Parnes, N. Strominger, S. Silver, and J. C. Goldstein
Alternate Innervations of Facial Musculature
Arch Otolaryngol Head Neck Surg, July 1, 1982; 108(7): 418 - 421.
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R. Wiley, W. Blessing, and D. Reis
Suicide transport: destruction of neurons by retrograde transport of ricin, abrin, and modeccin
Science, May 21, 1982; 216(4548): 889 - 890.
[Abstract] [PDF]

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P. Goldman-Rakic and M. Schwartz
Interdigitation of contralateral and ipsilateral columnar projections to frontal association cortex in primates
Science, May 14, 1982; 216(4547): 755 - 757.
[Abstract] [PDF]

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S. McLoon
Alterations in precision of the crossed retinotectal projection during chick development
Science, March 12, 1982; 215(4538): 1418 - 1420.
[Abstract] [PDF]

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S David and A. Aguayo
Axonal elongation into peripheral nervous system "bridges" after central nervous system injury in adult rats
Science, November 20, 1981; 214(4523): 931 - 933.
[Abstract] [PDF]

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M Mayberg, R. Langer, N. Zervas, and M. Moskowitz
Perivascular meningeal projections from cat trigeminal ganglia: possible pathway for vascular headaches in man
Science, July 10, 1981; 213(4504): 228 - 230.
[Abstract] [PDF]

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S. Breedlove and A. Arnold
Hormone accumulation in a sexually dimorphic motor nucleus of the rat spinal cord
Science, October 31, 1980; 210(4469): 564 - 566.
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T. Brushart and M. Mesulam
Alteration in connections between muscle and anterior horn motoneurons after peripheral nerve repair
Science, May 9, 1980; 208(4444): 603 - 605.
[Abstract] [PDF]

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D. Kelley
Auditory and vocal nuclei in the frog brain concentrate sex hormones
Science, February 1, 1980; 207(4430): 553 - 555.
[Abstract] [PDF]

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K Kalil and T Reh
Regrowth of severed axons in the neonatal central nervous system: establishment of normal connections
Science, September 14, 1979; 205(4411): 1158 - 1161.
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P. Land and R. Lund
Development of the rat's uncrossed retinotectal pathway and its relation to plasticity studies
Science, August 17, 1979; 205(4407): 698 - 700.
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B. Vogt, D. Rosene, and D. Pandya
Thalamic and cortical afferents differentiate anterior from posterior cingulate cortex in the monkey
Science, April 13, 1979; 204(4389): 205 - 207.
[Abstract] [PDF]

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R. Carey, D Fitzpatrick, and I. Diamond
Thalamic projections to layer I of striate cortex shown by retrograde transport of horseradish peroxidase
Science, February 9, 1979; 203(4380): 556 - 559.
[Abstract] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
S. Millecamps, D. Nicolle, I. Ceballos-Picot, J. Mallet, and M. Barkats
Synaptic sprouting increases the uptake capacities of motoneurons in amyotrophic lateral sclerosis mice
PNAS, June 19, 2001; 98(13): 7582 - 7587.
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D. O. Frost, D. Boire, G. Gingras, and M. Ptito
Surgically created neural pathways mediate visual pattern discrimination
PNAS, September 26, 2000; 97(20): 11068 - 11073.
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