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- W3080552305 abstract "552 Cytotoxicity assays using 96-well microplates offer several advantages over colony-forming or dye-exclusion assays, including automation, rapid sample handling, the ability to change the medium, and conditions similar to standard cell cultures. These assays employ colorimetrically measured dyes, radioactive isotopes, or fluorochromes to detect viable cells. The MTT assay, which is based on the colorimetric measurement of a formazan derivative of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide that is formed by live cells, has been the most frequently used assay for in vitro cytotoxicity testing. However, the dynamic range of this assay, which is 2 logs, might limit its use for testing drug combinations to identify syngergistic interactions. The DIMSCAN cytototoxicity assay employs fluorescein diacetate (FDA), a dye that selectively accumulates in viable cells and is quantified by a semi-automatic digital imaging fluorescence microscope (Cytometry, 24:204, 1996). DIMSCAN measures relative numbers of viable cells by evaluating the FDA fluorescence intensity in each well after the elimination of background fluorescence by digital thresholding and eosin Y staining and has a 93% sensitivity in detecting a single viable cell in a microplate well. DIMSCAN provides a linear relationship over a 4 log dynamic range between the number of deposited viable cells and the fluorescence intensity measured and scans a 96-well microplate in less than 7 min. We compared the ability to detect drug synergy between the DIMSCAN and MTT assays using combinations of anticancer drugs that are known to be active against neuroblastoma in vitro and in vivo: melphalan (L-PAM) + buthionine sulfoximine (BSO) (Bone Marrow Transplant 30:135, 2002), and fenretinide (4-HPR) + safingol (SAF) (J Natl Cancer Inst, 92:1897, 2000). After incubating various concentrations of the drugs in fixed ratios with the CHLA-90 multi-drug-resistant neuroblastoma cell line for five days, the microplates were analyzed with DIMSCAN or MTT. Results are summarized in the following table, with Combination Index (CI) values: 1.0 = antagonism. These data demonstrate that the larger dynamic range of the DIMSCAN assay allows identification of synergistic drug combinations that were not evident when employing the MTT assay." @default.
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- W3080552305 date "2004-04-01" @default.
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- W3080552305 title "The DIMSCAN cytotoxicity assay, unlike the MTT assay, identifies syngergistic combinations of anticancer agents" @default.
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