Confocal Microscopy: Principles and Practices

Phillip N. Dean1

1 null, Livermore, California
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 2.8
DOI:  10.1002/0471142956.cy0208s05
Online Posting Date:  May, 2001
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

The author presents a well rounded and complete overview of the topic, covering optics, detectors, image collection, and analysis, and designed to take the uninitiated person through an excellent understanding of this very actively growing technology.

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Design
  • Types of Confocal Microscopes
  • Methods of Scanning
  • Applications
  • Practical Considerations
  • Future Developments
  • Figures
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Amos, W.B., White, J.G., and Fordham, M. 1987. Use of confocal imaging in the study of biological structures. Appl. Opt. 26:3239‐3243.
   Boyde, A. 1985. Stereoscopic images in confocal (tandem scanning) microscopy. Science 230:1270‐1272.
   Brakenhoff, G.J. and Visscher, K. 1990. Novel confocal imaging and visualization techniques. In Transactions of the Royal Microscopical Society, Vol. 1. (H.Y. Elder, ed.) pp. 247‐250. Adam Hilger, Bristol.
   Cavanagh, H.D., Petroll, W.M., and Jester, J.V. 1993. The application of confocal microscopy to the study of living systems. Neurosci.Biobehav. Rev. 17.4:483‐498.
  D Cogswell, C.J. and Sheppard, C.J.R. 1990. Confocal brightfield imaging techniques using an on‐axis scanning optical microscope. In Confocal Microscopy (T. Wilson, ed.) pp. 213‐243 Academic Press, London.
   Cogswell, C.J. and Sheppard, C.J.R. 1992. Confocal differential interference contrast (DIC) microscopy: Including a theoretical analysis of conventional and confocal DIC imaging. J. Microscopy 165:81‐101.
   Denk, W., Strickler, J.H., and Webb, W.W. 1990. Two‐photon laser scanning fluorescence microscopy. Science 248:73‐76.
   Draaijer, A. and Houpt, P.M. 1987. A real‐time confocal laser scanning microscope (CLSM). Proc. SPIE 809:85‐88.
   Florijn, R.J., Slats, J., Tanke, H.J., and Rapp, A.K. 1995. Analysis of antifading reagents for fluorescence microscopy. Cytometry 19:177‐182.
   Kino, G.S. 1989. Efficiency in Nipkow disk microscopes. In The Handbook of Biological Confocal Microscopy (J. Pawley, ed.) pp.93‐97. IMR Press, Madison, Wis.
   Matsumoto, B. 1993. Cell Biological Applications of Confocal Microscopy. Methods in Cell Biology,Volume 38. Academic Press, New York.
   Minsky, M. 1957. Microscopy Apparatus. U.S. Patent no. 3013467 (awarded 1961).
   Minsky, M. 1988. Memoir on inventing the confocal scanning microscope. Scanning 10:128‐138.
   Nipkow, P. 1984. German Patent no. 30105.
   Petran, M., Hadravsky, M., Egger, D., and Galambos, R. 1968. Tandem‐scanning reflected light microscope. J. Opt. Soc. Amer. 58:661‐664.
   Wilson, T. (ed.) 1990. Confocal Microscopy. Academic Press, London.
Key References
   Elder, H.Y. (ed.) 1990. Transactions of the Royal Microscopical Society, Vol. 1. Adam Hilger, Bristol.
  Contains papers presented at Micro 90, a conference of the Society held in 1990 at Shortlands, Hammersmith, London. The 21 chapters cover virtually all aspects of microscopy. In particular, it contains excellent papers on the physics and configuration of confocal microscopes, as well as on methods of staining and preparation of samples and on three‐dimensional image processing and applications.
   Matsumoto, B. 1993. See
  Contains 13 chapters with extensive discussions of a variety of applications that utilize confocal microscopy.
   Pawley, J. (ed.) 1989. The Handbook of Biological Confocal Microscopy. IMR Press, Madison, Wis.
  Consists of 19 papers covering the practical aspects of confocal microscopy; an excellent general reference to the topic.
   Wilson, T. 1990. See
  Contains 15 chapters covering all aspects of confocal microscopy, primarily from the theoretical point of view.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library