This Article Free Full Text (Free PDF) Free All Versions of this Article: 1087057106289406v1 11/6/586    most recent References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (6) Citing Articles via Google Scholar Google Scholar Articles by Gasparri, F. Articles by Galvani, A. Search for Related Content PubMed PubMed Citation Articles by Gasparri, F. Articles by Galvani, A. Social Bookmarking                   What's this?

Multiparametric Cell Cycle Analysis by Automated Microscopy

Biology Department, Oncology, Nerviano Medical Sciences, Nerviano, Italy

Paolo Cappella

Biology Department, Oncology, Nerviano Medical Sciences, Nerviano, Italy

Arturo Galvani

Biology Department, Oncology, Nerviano Medical Sciences, Nerviano, Italy

Cell cycle analysis using flow cytometry (FC) to measure cellular DNA content is a common procedure in drug mechanism of action studies. Although this technique lends itself readily to cell lines that grow in suspension, adherent cell cultures must be resuspended in a cumbersome and potentially invasive procedure that normally involves trypsinization and mechanical agitation of monolayer cultures. High-content analysis (HCA), an automated microscopy-based technology, is well suited to analysis of monolayer cell cultures but provides intrinsically less accurate determination of cellular DNA content than does FC and thus is not the method of choice for cell cycle analysis. Using Cellomics’s ArrayScan^TM reader, the authors have developed a 4-color multiparametric HCA approach for cell cycle analysis of adherent cells based on detection of DNA content (4,6-diamidino-2-phenylindole [DAPI] fluorescence), together with the known cell cycle markers bromo-2-deoxyuridine (BrdU) incorporation, cyclin B1 expression, and histone H3 (Ser28) phosphorylation within a single cell population. Considering all 4 markers together, a reliable and accurate quantification of cell cycle phases was possible, as compared with flow cytometric analysis. Using this assay, specific cell cycle blocks induced by treatment with thymidine, paclitaxel, or nocodazole as test drugs were easily monitored in adherent cultures of U-2 OS osteosarcoma cells.

Key Words: Cell-based assays • ArrayScan^TM • high-content analysis • cell-cycle • U-2 OS

CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C. O'Brien, G. Cavet, A. Pandita, X. Hu, L. Haydu, S. Mohan, K. Toy, C. S. Rivers, Z. Modrusan, L. C. Amler, et al. Functional Genomics Identifies ABCC3 as a Mediator of Taxane Resistance in HER2-Amplified Breast Cancer Cancer Res., July 1, 2008; 68(13): 5380 - 5389. [Abstract] [Full Text] [PDF]

				L. Barbe, E. Lundberg, P. Oksvold, A. Stenius, E. Lewin, E. Bjorling, A. Asplund, F. Ponten, H. Brismar, M. Uhlen, et al. Toward a Confocal Subcellular Atlas of the Human Proteome Mol. Cell. Proteomics, March 1, 2008; 7(3): 499 - 508. [Abstract] [Full Text] [PDF]

				A. Sartore-Bianchi, F. Gasparri, A. Galvani, L. Nici, J. W. Darnowski, D. Barbone, D. A. Fennell, G. Gaudino, C. Porta, and L. Mutti Bortezomib Inhibits Nuclear Factor-{kappa}B Dependent Survival and Has Potent In vivo Activity in Mesothelioma Clin. Cancer Res., October 1, 2007; 13(19): 5942 - 5951. [Abstract] [Full Text] [PDF]