Screening for apoptosis--classical and emerging techniques.
There has been a rapid development of cell-based assays and screening methods to identify promising apoptosis-inducing drug candidates for the treatment of cancer. Distinguishing between the complex processes involved in apoptosis and other forms of cell death requires information on both biochemical and morphological processes in the cell. Traditionally, many assays have been limited to measuring, for example, caspase activity using fluorogenic substrates. However, these screening assays provide only limited information on the complex processes involved in apoptosis. In this review we describe some of the available apoptosis assays amenable to high-throughput screening. In particular, image-based high-content screening assays to evaluate multiple biochemical and morphological parameters in apoptotic cells are described. Through combining the imaging of cells in microtiter plates with powerful image analysis algorithms, one can acquire deeper knowledge on multiple biochemical or morphological pathways at the single-cell level at an early stage in the development of novel anti-cancer drugs.
Lövborg
Henrik
H
Department of Medical Sciences, Division of Clinical Pharmacology, University Hospital, Uppsala University, Sweden. henrik.lovborg@medsci.uu.se
Gullbo
Joachim
J
Larsson
Rolf
R
D000818:Animals; D016588:Anticarcinogenic Agents; D017209:Apoptosis / Q000276:immunology; D020169:Caspases / Q000378:metabolism; D003592:Cytophotometry / Q000379:methods*; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D019044:Image Cytometry / Q000379:methods; D007091:Image Processing, Computer-Assisted; D007118:Immunoassay; D008856:Microscopy, Fluorescence
D016588:Anticarcinogenic Agents; D020169:Caspases