Stabilization of tetramethylbenzidine (TMB) reaction product at the electron microscopic level by ammonium molybdate

Carl F. Marfurt, Dennis F. Turner, Catherine E. Adams

Research output: Contribution to journalArticle

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Abstract

The ability to use the tetramethylbenzidine (TMB) method for studying neuronal connections at the electron microscopic level is often difficult because the conditions of osmification and dehydration used in processing the tissue may result in significant loss and/or decreased electron density of the reaction product. In the present study, we report that stabilization of TMB reaction product with 5% ammonium molybdate (AM) prior to osmificating the tissue results in the formation of TMB-AM crystals that are many times more electron dense and resistant to ethanol extraction than non-stabilized TMB crystals. The nature of the chemical interaction that underlies the stabilization of TMB by AM is uncertain, but it may involve the formation of an insoluble salt between molybdic ions and the TMB polymer. The use of this simple procedure increases the sensitivity of the TMB procedure at the electron microscopic level and may be used to label neuronal pathways in the peripheral and central nervous systems with equal success.

Original languageEnglish (US)
Pages (from-to)215-223
Number of pages9
JournalJournal of Neuroscience Methods
Volume25
Issue number3
DOIs
StatePublished - Oct 1988

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Keywords

  • Ammonium molybdate
  • Electron microscopy
  • Horseradish peroxidase
  • Tetramethylbenzidine (TMB) method

ASJC Scopus subject areas

  • Neuroscience(all)

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