An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells

Paula Szalai; Nikolai Engedal

doi:10.21769/BioProtoc.2835

An Image-based Assay for High-throughput Analysis of Cell Proliferation and Cell Death of Adherent Cells

Bio Protoc. 2018 May 5;8(9):e2835. doi: 10.21769/BioProtoc.2835.

Authors

Paula Szalai¹, Nikolai Engedal¹

Affiliation

¹ The Autophagy Team, Centre for Molecular Medicine Norway (NCMM), Nordic EMBL Partnership, University of Oslo, N-0318 Oslo, Norway.

Abstract

In this protocol, we describe a method to monitor cell proliferation and death by live-cell imaging of propidium iodide (PI)-stained adherent mammalian cells. PI is widely used to assess cell death. However, it is usually used in end-point assays. Recently, we implemented the use of PI for real-time cell death assessment by automated imaging. Cells are seeded in a 96-well format, and after attachment, the treatments are added directly to the wells together with PI. Thereafter, cells are subjected to automated time-lapse imaging and quantification by computer software. Combined analyses of phase-contrast and fluorescence images allow assessment of treatment effects on cell proliferation as well as the extent and kinetics of cell death.

Keywords: Cell death; Cell proliferation; Live-cell imaging; Propidium iodide; Thapsigargin; Time-lapse imaging.