SpringerProtocols: Abstract: Automated Live Microscopy to Study Mitotic Gene Function in Fluorescent Reporter Cell Lines

Automated Live Microscopy to Study Mitotic Gene Function in Fluorescent Reporter Cell Lines

By: Michael H.A. Schmitz¹, Daniel W. Gerlich¹

Abstract

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Fluorescence live microscopy is a powerful technique to study complex cellular dynamics such as cell division. The availability of fluorescent markers based on GFP fusion proteins for virtually any cellular structure allows efficient visualization of specific processes, and the combination of different fluorophores can be used to study their coordination. In this chapter, we present methods for automated live cell microscopy to study mitotic gene function systematically and in high throughput. In particular, we provide protocols for efficient generation of fluorescent reporter cell lines stably expressing combinations of cellular markers, and provide detailed guidelines for optimizing imaging protocols for automated long-term live microscopy.

Affiliation(s): (1) Institute of Biochemistry, ETH Zurich, Zurich, Switzerland

Book Title: Mitosis: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 545 | Pub. Date: Aug-01-2009 | Page Range: 113-134 | DOI: 10.1007/978-1-60327-993-2_7

Subject: Cell Biology

Key Words: Automated live-cell imaging - multidimensional microscopy - laser scanning confocal microscopy - widefield epifluorescence microscopy - mitosis - Green Fluorescent Protein (GFP) - stably expressing reporter cell lines

References

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