A novel application of the fluorescent dye bis-ANS for labeling neurons in acute brain slices

Emese Mozes, Akos Hunya, Aniko Toth, Ferhan Ayaydin, Botond Penke, Zsolt L. Datki

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The cell-impermeant oligomer-(e.g. beta-amyloid-, or tubulin-) specific fluorescent dye, bis-ANS (4,4'-bis-1-anilinonaphtalene-8-sulfonate), was successfully used for labeling mechanically damaged but still viable neuron bodies, neurites and neurite cross sections in acute brain slices.Acute hippocampal brain slices of rats were co-stained with bis-ANS and the cell-impermeant, DNA-specific dye propidium iodide (PI) and were then analyzed using fluorescence and confocal microscopes. Both the neuron bodies and the neurites were found to exhibit increased fluorescence intensities, suggesting that using this method they can be detected more easily. In addition, bis-ANS showed good region - but not cell specific co-localization with the neuron-specific fluorescent dye Dil (1,1'-Dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate). These two dyes label different neuronal structures: Dil binds specifically to intact cell membranes while bis-ANS can enter cells with compromised cell membranes and then stain the microtubules in the cytoplasm. For a quick (10min) staining of acute brain slices with bis-ANS both HEPES and NaHCO 3 were needed in order to achieve high signal intensity.Labeling with bis-ANS fluorescent dye is an easy method for imaging the neuronal structures on the surface of acute brain slices.

Original languageEnglish
Pages (from-to)217-221
Number of pages5
JournalBrain Research Bulletin
Volume86
Issue number3-4
DOIs
Publication statusPublished - Oct 10 2011

Keywords

  • Acute slice
  • Bis-ANS
  • Fluorescence
  • Hippocampus
  • Neuron
  • SH-SY5Y

ASJC Scopus subject areas

  • Neuroscience(all)

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