Rapid assay for quantitative measurement of apoptosis in cultured cells and brain tissue

Bryce Patrick Portier, Diana Carolina Ferrari, Giulio Taglialatela

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Analysis of apoptosis in brain tissue following ischemia, hypoxia, or oxidative stress has technical limitations. The use of counting cells displaying apoptotic morphology is time intensive, vulnerable to sampling errors, and suffers from low numbers of total recorded events. Other cell death assays such as agarose gel analysis of DNA fragmentation, TUNEL, or ELISA are time intensive, limited to a single endpoint measure, and can be technically difficult to perform or reproduce. To overcome these limitations, we set out to develop a technique using flow cytometry to measure apoptosis based on the physical properties of light scatter produced from isolated nuclei. This dye/marker free approach would bypass many of the inherent encumbrances and reproducibility problems found in other apoptosis assays. Here we demonstrate that this new technique, using flow cytometry performed on isolated nuclei, allows rapid quantitation of apoptosis in a variety of brain tissues without the need for intercalating dyes or fluorescent markers. We conclude that this technique significantly improves currently available protocols to quantify apoptosis from tissue and offers the possibility to perform additional analysis on the same population of nuclei via downstream assays.

Original languageEnglish (US)
Pages (from-to)134-142
Number of pages9
JournalJournal of Neuroscience Methods
Volume155
Issue number1
DOIs
StatePublished - Jul 15 2006

Keywords

  • Apoptosis
  • Brain
  • Flow cytometry
  • Nuclei

ASJC Scopus subject areas

  • Neuroscience(all)

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