Flow cytometry analysis of gap junction-mediated Cel-Cell communication: Advantages and pitfalls

Paula Candida Fonseca, Oscar Kenji Nihei, Wilson Savino, David C. Spray, Luiz Anastacio Alves

Research output: Contribution to journalReview article

29 Scopus citations

Abstract

Background: Since the first morphological description of the gap junctions use electron microscopy, a considerable number of techniques has been introduced to evaluate gap junction channel functionality, many of which use dye transfer techniques, such as dye injection and fluorescent dye transfer, analyzed by flow cytometry. Methods: To analyze dye transfer, generally one population of cells is incubated with calcein-AM (0.5 μM) for 30 min at 37°C, and the other population was incubated with the lipophilic dye DiIC18 (3) (10 μM) for 1h at 37°C; after incubation, these cells were washed five times with PBS and cocultured for different times, and then the dye transfer was analyzed by flow cytometry. Results: In this short overview, we focus on some advantages and disadvantages of flow cytometry as a technique to investigate gap junction-mediated intercellular communication (GJIC). In addition, we point out some technical pitfalls that we have encountered when applying this technique to study gap junctions in immune system cells. Conclusions: Analysis of fluorescent dye transfer by flow cytometry is a useful tool to investigate GJIC. However, some points must be taken into consideration before using this methodology, which are discussed herein.

Original languageEnglish (US)
Pages (from-to)487-493
Number of pages7
JournalCytometry Part A
Volume69
Issue number6
DOIs
StatePublished - Jan 1 2006

Keywords

  • Flow cytometry
  • Gap junctions
  • Intercellular communication
  • Lymphocytes

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

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