title: High-resolution cytometry for high-content cell cycle analysis.
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.
authors.affiliation: Department of Experimental Oncology, European Institute of Oncology, Milan, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Furia
authors.forename: Piergiuseppe
authors.initials: P
authors.lastname: Pelicci
authors.forename: Mario
authors.initials: M
authors.lastname: Faretta
mesh_terms: D000818:Animals; D001331:Automation / Q000295:instrumentation / Q000379:methods; D002453:Cell Cycle; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods; D008853:Microscopy / Q000295:instrumentation / Q000379:methods
keywords: S phase; automated microscopy; cell cycle; high resolution; high-content analysis; image cytometry; wide-field microscopy