Simultaneous observation of fluorescent retrogradely labeled neurons and the immunofluorescently labeled fibers apposing them using Fluoro-Gold and antisera labeled with the blue fluorochrome 7-amino-4-methylcoumarin-3-acetic acid (AMCA)

Martin W Wessendorf, Nathan M. Appel, Robert Elde

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

In this paper we describe a method allowing the simultaneous observation of fluorescent retrogradely labeled neurons and the immunofluorescently labeled fibers which appose them. This technique employs the fluorescent dye Fluoro-Gold to label neurons retrogradely, and 7-amino-4-methylcoumarin-3-acetic-acid (AMCA) conjugated to a secondary antiserum as a label for fluorescence immunohistochemistry. Both Fluoro-Gold and AMCA are excited by near-ultraviolet (UV) light, but under UV excitation Fluoro-Gold appears yellow and AMCA appears blue. Thus the two fluorochromes are both visible under a single condition of illumination and can be readily distinguished by color. This allows quick and accurate determination of whether or not immunofluorescent fibers appose retrogradely labeled neurons.

Original languageEnglish (US)
Pages (from-to)121-126
Number of pages6
JournalNeuroscience Letters
Volume82
Issue number2
DOIs
StatePublished - Nov 23 1987

Keywords

  • 7-Amino-4-methylcoumarin-3-acetic-acid
  • AMCA
  • Fluoro-Gold
  • Immunofluorescence
  • Intermediolateral cell column
  • Retrograde labeling
  • Serotonin
  • Substance P

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