Protocol abstract


Nature Protocols 3, 1364 - 1369 (2008)
Published online: 31 July 2008 | doi:10.1038/nprot.2008.114

Subject Categories: Biochemistry and protein analysis | Pharmacology and toxicology

The fluorometric microculture cytotoxicity assay

Elin Lindhagen1, Peter Nygren2 & Rolf Larsson1


The fluorometric microculture cytotoxicity assay (FMCA) is a nonclonogenic microplate-based cell viability assay used for measurement of the cytotoxic and/or cytostatic effect of different compounds in vitro. The assay is based on hydrolysis of the probe, fluorescein diacetate (FDA) by esterases in cells with intact plasma membranes. The assay is available as both a semiautomated 96-well plate setup and a 384-well plate version fully adaptable to robotics. Experimental plates are prepared with a small amount of drug solution and can be stored frozen. Cells are seeded on the plates and cell viability is evaluated after 72 h. The protocol described here is applicable both for cell lines and freshly prepared tumor cells from patients and is suitable both for screening in drug development and as a basis for a predictive test for individualization of anticancer drug therapy.

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  1. Division of Clinical Pharmacology, Department of Medical Sciences, Uppsala University, Uppsala University Hospital, SE-751 85 Uppsala, Sweden.
  2. Division of Oncology, Department of Oncology, Radiology and Clinical Immunology, Uppsala University, Uppsala University Hospital, SE-751 85 Uppsala, Sweden.

Correspondence to: Elin Lindhagen1 e-mail: elin.lindhagen@medsci.uu.se