High‐Resolution Cytometry for High‐Content Cell Cycle Analysis. Issue 1 (1st October 2014)

• Record Type: Journal Article
• Authors: Furia, Laura; Pelicci, Piergiuseppe; Faretta, Mario
• Editors: Robinson, J. Paul; Darzynkiewicz, Zbigniew; Hoffman, Robert; Nolan, John P.; Rabinovitch, Peter S.; Watkins, Simon
• Is Part Of: Current protocols in cytometry. Volume 70:Issue 1(2014)
• Page Start: 7.41.1
• Page End: 7.41.15
• Publication Date: 2014-10-01
• Subjects: image cytometry -- cell cycle -- S phase -- high‐content analysis -- high resolution -- automated microscopy -- wide‐field microscopy; Cytology -- Laboratory manuals; Flow cytometry -- Laboratory manuals; Cell separation -- Laboratory manuals; Molecular biology -- Laboratory manuals; Flow Cytometry -- methods; Image Cytometry -- methods; Cell Separation -- methods; Cytological Techniques; Molecular Biology -- methods; Cell separation; Cytology; Flow cytometry; Molecular biology; Laboratory Manuals; Laboratory manuals; 571.6
• DOI: 10.1002/0471142956.cy0741s70
• Languages: English
• ISSNs: 1934-9297

Abstract

Abstract: One of the major limitations of flow cytometry (FCM) is the absence of an intracellular view. Automated microscopy and image analysis, together with technological developments, led to new approaches in cytometry that bypass the above limitation, introducing high resolution, high content, and large statistical sampling. However, few attempts have been made, until now, to translate the wide repertoire of FCM assays into high‐content image screening. This unit describes the implementation of an acquisition and analysis protocol for evaluation of the cell cycle by automated microscopy. The approach grants the possibility to perform simultaneous analysis of a high number of different parameters. A large part of this unit is devoted to the description of hardware features that can optimize the recorded information together with the acquisition and analysis procedures employed to produce good‐quality data. Curr. Protoc. Cytom . 70:7.41.1‐7.41.15. © 2014 by John Wiley & Sons, Inc.