title: Confocal microscopy for high-resolution and high-content analysis of the cell cycle.
abstract: Optical fluorescence microscopy offers a wide range of technological solutions to address many questions in biomedical research. Spatial resolution has been greatly improved by the use of confocal microscopes, providing a 3-D analysis of the intracellular space. Automation has contributed to make confocal analysis available for high-content image cytometry studies. However, the storage, browsing, and analysis of the amount of data generated can challenge the feasibility of such studies. Presented in this chapter is a multistep acquisition and analysis protocol that can bypass such difficulties by an analysis-driven data collection. Cell-cycle analysis of low-resolution data can be employed to select cell populations of interest that can then be imaged at extremely high resolution and subjected to high-content analysis.
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; D002453:Cell Cycle; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation* / Q000379:methods*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods
keywords: S phase; automated microscopy; cell cycle; confocal microscopy; high content; high resolution; image cytometry