A Morphological identification cell cytotoxicity assay using cytoplasm-localized fluorescent probe (CLFP) to distinguish living and dead cells

Biochem Biophys Res Commun. 2017 Jan 8;482(2):257-263. doi: 10.1016/j.bbrc.2016.09.169. Epub 2016 Nov 11.

Abstract

Cell cytotoxicity assays include cell activity assays and morphological identification assays. Currently, all frequently used cytotoxicity assays belong to cell activity assays but suffer from detection limitations. Morphological identification of cell death remains as the gold standard, although the method is difficult to scale up. At present there is no generally accepted morphological identification based cell cytotoxicity assay. In this study, we applied previous developed cell cytoplasm-localized fluorescent probe (CLFP) to display cell morphologies. Under fluorescence microscopy, the fluorescence morphology and intensity of living cells are distinct from dead cells. Based on these characters we extracted the images of living cells from series of samples via computational analysis. Thus, a novel cell morphological identification cytotoxicity assay (CLFP assay) is developed. The performance of the CLFP assay was similar to cell activity assay (MTT assay), but the accuracy of the CLFP assay was superior when measuring the cytotoxicity of active compounds.

Keywords: Cell death; Cell morphological identification; Computational analysis; Cytoplasm-localized fluorescent probe; Cytotoxicity assay.

MeSH terms

  • Apoptosis / drug effects*
  • Apoptosis / physiology
  • Biological Assay / methods
  • Cell Size / drug effects
  • Cell Survival / drug effects*
  • Cell Survival / physiology
  • Cytoplasm / chemistry
  • Cytotoxins / administration & dosage*
  • Fluorescent Dyes / chemistry*
  • Microscopy, Fluorescence / methods*
  • Toxicity Tests / methods*

Substances

  • Cytotoxins
  • Fluorescent Dyes