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First published online March 4, 2009
doi: 10.1242/10.1242/jcs.033837

Journal of Cell Science 122, 753-767 (2009)
Published by The Company of Biologists 2009
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Commentary

Live-cell microscopy – tips and tools

Melanie M. Frigault1, Judith Lacoste2,3, Jody L. Swift4 and Claire M. Brown2,*

1 Molecular Oncology Group, McGill University Cancer Centre, Montreal, Canada
2 McGill University Life Sciences Complex Imaging Facility, Department of Biochemistry, Montreal, Canada
3 Cell Imaging and Analysis Network, Department of Biology, McGill University, Montreal, Canada
4 Department of Chemistry, McGill University, Montreal, Canada

* Author for correspondence (e-mail: claire.brown{at}mcgill.ca)

Imaging of living cells and tissue is now common in many fields of the life and physical sciences, and is instrumental in revealing a great deal about cellular dynamics and function. It is crucial when performing such experiments that cell viability is at the forefront of any measurement to ensure that the physiological and biological processes that are under investigation are not altered in any way. Many cells and tissues are not normally exposed to light during their life cycle, so it is important for microscopy applications to minimize light exposure, which can cause phototoxicity. To ensure minimal light exposure, it is crucial that microscope systems are optimized to collect as much light as possible. This can be achieved using superior-quality optical components and state-of-the-art detectors. This Commentary discusses how to set up a suitable environment on the microscope stage to maintain living cells. There is also a focus on general and imaging-platform-specific ways to optimize the efficiency of light throughput and detection. With an efficient optical microscope and a good detector, the light exposure can be minimized during live-cell imaging, thus minimizing phototoxicity and maintaining cell viability. Brief suggestions for useful microscope accessories as well as available fluorescence tools are also presented. Finally, a flow chart is provided to assist readers in choosing the appropriate imaging platform for their experimental systems.

Key words: CLSM, Multi-photon, TIRF, Live-cell imaging, Spinning disk


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© The Company of Biologists Ltd 2009