Video-Rate Confocal Microscopy

  • Roger Y. Tsien
  • Brian J. Bacskai
Chapter

Abstract

Most current confocal laser-scanning microscopes require <0.1 sec, more typically ~1 sec, to complete a single full scan of the field of view. Such speeds are considerably slower than conventional video displays or the psychophysical flicker-fusion frequency, so they give the visual impression of discontinuous image acquisition, rather than a smoothly evolving scene. This speed limitation is imposed by the scanning rates of conventional scanners based on linear servo galvanometers, not from any inherent feature of the confocal principle. Scanning at video rate has many advantages. The most obvious but not necessarily the most important is the psychophysical appearance of smooth motion or evolution, which greatly aids the user to locate and focus on regions of interest. Many biochemical signals inside living cells, particularly membrane potential and cytosolic free Ca2+and Na+, undergo changes in the time scale of seconds down to tenths of milliseconds. Such signals are especially important in neurobiology because neurons rely on such biochemistry to perform their special function of fast information processing.

Keywords

Image Processor Optical Layout Color Insert Neonatal Cardiac Myocytes Polarize Beamsplitter 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Roger Y. Tsien
    • 1
  • Brian J. Bacskai
    • 1
  1. 1.Howard Hughes Medical Institute and Department of Pharmacology 0647University of California, San DiegoLa JollaUSA

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