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

Bio-protocol (2018-05-05)
Paula Szalai, Nikolai Engedal
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Thapsigargin, ≥98% (HPLC), solid film
Sigma-Aldrich
Dimethyl sulfoxide, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
Sigma-Aldrich
Propidium Iodide - CAS 25535-16-4 - Calbiochem, Membrane impermeable DNA intercalator.