Scrape Loading/Dye Transfer Assay

Methods Mol Biol. 2016:1437:133-44. doi: 10.1007/978-1-4939-3664-9_9.

Abstract

The scrape loading/dye transfer (SL/DT) technique is a simple functional assay for the simultaneous assessment of gap junctional intercellular communication (GJIC) in a large population of cells. The equipment needs are minimal and are typically met in standard cell biology labs, and SL/DT is the simplest and quickest of all the assays that measure GJIC. This assay has also been adapted for in vivo studies. The SL/DT assay is also conducive to a high-throughput setup with automated fluorescence microscopy imaging and analysis to elucidate more samples in shorter time, and hence can serve a broad range of in vitro pharmacological and toxicological needs.

Keywords: Dye coupling; Dye transfer; Ex vivo assessment; Gap junctional intercellular communication assessment; High throughput; In vitro assay; Incision loading; Lucifer Yellow; Scalpel loading; Scrape loading; Tracers.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Communication*
  • Cells, Cultured
  • Dextrans / metabolism
  • Fluorescent Dyes / metabolism*
  • Gap Junctions / metabolism*
  • Gap Junctions / ultrastructure
  • Isoquinolines / metabolism
  • Male
  • Mice
  • Microscopy, Fluorescence / methods
  • Rhodamines / metabolism
  • Sertoli Cells

Substances

  • Dextrans
  • Fluorescent Dyes
  • Isoquinolines
  • Rhodamines
  • rhodamine dextran
  • lucifer yellow