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Microscopy Reference Library

Spinning Disk Confocal Microscopy

Spinning disk confocal microscopy is rapidly emerging as the technique of choice for investigation of dynamics in living cells. Modern commercial instruments and high-performance camera systems are capable of providing high acquisition speeds with acceptable contrast and minimal photobleaching at the low light levels available with this technique. The references listed in this section point to review articles that should provide the starting point for a thorough understanding of spinning disk confocal microscopy.

Gräf, R., Rietdorf, J. and Zimmermann, T.

Live Cell Spinning Disk Microscopy.  Advances in Biochemical Engineering/Biotechnology 95: 57-75 (2005).  An excellent review of spinning disk microscopy with emphasis on live-cell imaging. The authors discuss the benefits of spinning disk systems over laser scanning confocal microscopes, including image acquisition rates, photobleaching, multichannel imaging, and optical sectioning. Several examples of biological imaging applications are described.

Wang, E., Babbey, C. M. and Dunn, K. W.

Performance comparison between the high-speed Yokogawa spinning disk confocal system and single-point scanning confocal systems.  Journal of Microscopy 218: 148-159 (2005).  A comparative investigation of spinning disk and laser scanning confocal microscopy using signal-to-noise and photobleaching as an index of system efficiency. Included are detail discussions of instrument configuration, resolution, photobleaching sensitivity, and saturation effects.

Adams, M. C., Salmon, W. C., Gupton, S. L., Cohan, C. S., Wittmann, T., Prigozhina, N. and Waterman-Storer, C. M.

A high-speed multispectral spinning-disk confocal microscope system for fluorescence speckle microscopy of living cells.  Methods 29: 29-41 (2003).  The authors describe a multispectral spinning disk instrument designed to analyze rapid dynamic processes in living cells. Included are a description of the instrumental configuration parameters, detector sensitivity, laser illumination source, capture and analysis software, and examples of biological applications.

Nakano, A.

Spinning-disk confocal microscopy: a cutting-edge tool for imaging membrane traffic.  Cell Structure and Function 27: 349-355 (2002).  One of the first descriptions of the Yokogawa Nipkow spinning disk microscope configuration that incorporates a twin disk system, with one disk being equipped with microlenses over the pinholes. The author describes sensitivity, speed, resolution, and detector requirements, as well as providing several examples of biological applications.

Inoué, S. and Inoué, T.

Direct-view high-speed confocal scanner: the CSU-10.  Methods in Cell Biology 70: 87-127 (2002).  The authors describe the design characteristics of the Yokogawa CSU-10 spinning disk confocal system and provide an overview of confocal microscopes. Also discussed are image processing, mechanical and optical performance of the CSU-10, and several imaging examples of pollen, mitochondria, heart muscle cells, and yeast microtubules.

Kozubek, M., Matula, P., Matula, P. and Kozubek, S.

Automated acquisition and processing of multidimensional image data in confocal in vivo microscopy.  Microscopy Research and Technique 64: 164-175 (2004).  Extending spinning disk microscope to live animal investigations, the authors describe how to optimize image acquisition and object tracking analysis for in vivo applications. The paper also provides an excellent comparison between commercial microscopes that are useful for these investigations.

Coates, C. G., Denvir, D. J., McHale, N. G., Thornbury, K. D. and Hollywood, M. A.

Optimizing low-light microscopy with a back-illuminated electron multiplying charge-coupled device: enhanced sensitivity, speed, and resolution.  Journal of Biomedical Optics 9: 1244-1252 (2004).  The authors introduce the application of electron multiplying digital cameras (EMCCDs) to spinning disk microscopy with an extensive discussion of sensitivity and resolution. Also described are a number of examples of imaging living cells at very low light levels using an EMCCD coupled to a spinning disk microscope.

Watson, T. F., Juskaitis, R. and Wilson, T.

New imaging modes for lenslet-array tandem scanning microscopes.  Journal of Microscopy 205: 209-212 (2002).  The application of non-coherent light sources and reflected illumination to spinning disk microscopy is examined in this original research report. Included are a schematic diagram of the microscope configuration, a discussion of resolution, and several example applications.

Xiao, G. Q., Corle, T. R. and Kino, G. S.

Real-time confocal scanning optical microscope.  Applied Physics Letters 53: 716-718 (1988).  One of the first publications describing a real-time spinning disk confocal microscope designed for reflected light specimens. The authors describe the microscope configuration, disk parameters, resolution, and optical sectioning. Also included are images of an integrated circuit obtained at different focal planes.

Bewersdorf, J., Pick, R. and Hell, S. W.

Multifocal multiphoton microscopy  Optics Letters 23: 655-657 (1998).  A description of multiphoton imaging using a spinning disk confocal microscope coupled to a titanium:sapphire laser system. Included is a discussion of resolution and several examples demonstrating the imaging of biological specimens.