1. Academic Validation
  2. Detection of cell aggregation and altered cell viability by automated label-free video microscopy: a promising alternative to endpoint viability assays in high-throughput screening

Detection of cell aggregation and altered cell viability by automated label-free video microscopy: a promising alternative to endpoint viability assays in high-throughput screening

  • J Biomol Screen. 2015 Mar;20(3):372-81. doi: 10.1177/1087057114562158.
Obaid Aftab 1 Mårten Fryknäs 1 Ulf Hammerling 1 Rolf Larsson 1 Mats G Gustafsson 2
Affiliations

Affiliations

  • 1 Department of Medical Sciences, Cancer Pharmacology and Computational Medicine, Uppsala University, Uppsala Academic Hospital, Uppsala, Sweden.
  • 2 Department of Medical Sciences, Cancer Pharmacology and Computational Medicine, Uppsala University, Uppsala Academic Hospital, Uppsala, Sweden Mats.Gustafsson@medsci.uu.se.
Abstract

Automated phase-contrast video microscopy now makes it feasible to monitor a high-throughput (HT) screening experiment in a 384-well microtiter plate format by collecting one time-lapse video per well. Being a very cost-effective and label-free monitoring method, its potential as an alternative to cell viability assays was evaluated. Three simple morphology feature extraction and comparison algorithms were developed and implemented for analysis of differentially time-evolving morphologies (DTEMs) monitored in phase-contrast microscopy videos. The most promising layout, pixel histogram hierarchy comparison (PHHC), was able to detect several compounds that did not induce any significant change in cell viability, but made the cell population appear as spheroidal cell aggregates. According to recent reports, all these compounds seem to be involved in inhibition of platelet-derived growth factor receptor (PDGFR) signaling. Thus, automated quantification of DTEM (AQDTEM) holds strong promise as an alternative or complement to viability assays in HT in vitro screening of chemical compounds.

Keywords

PDGFR signaling; cell aggregation; differentially time evolving morphologies; high-throughput screening (HTS); phase contrast microscopy; time-lapse microscopy; video microscopy.

Figures
Products