Real-Time Stereo (3D) Confocal Microscopy

  • G. J. Brakenhoff
  • K. Visscher


Three-dimensional (3D) microscopy requires the collection of data over a certain volume in the specimen, followed by a suitable two-dimensional (2D) visualization procedure in which the desired image is produced. In holography, nature offers us a potential pathway by which the 3D structure of an object can be observed directly through wavefront reconstruction techniques. However, the slow time response of holographic recording media prevents real-time applications of this technique. In addition, while in principle this approach might work in reflection, holographic fluorescence images are impossible because emitted fluorescence radiation is incoherent.


Confocal Image Emission Flux Emission Capability Readout Noise Camera Plane 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Agard, D.A., Hiraoka, Y., Shaw, P., and Sedat, J.W., 1989, Microscopy in three dimensions, Methods Cell Biol. 30:353–377.PubMedCrossRefGoogle Scholar
  2. Brakenhoff, G.J., 1979, Imaging modes in confocal scanning light microscopy (CSLM), J.Microsc. 117(2):233–242.CrossRefGoogle Scholar
  3. Brakenhoff, G.J., and van der Voort, H.T.M., 1989, Illumination and detection strategies for confocal microscopy, Proc. SPIE 1139:117–120.CrossRefGoogle Scholar
  4. Brakenhoff, G.J., and Visscher, K., 1992, Confocal imaging with bilateral scanning and array detectors, J. Microsc. 165:139–146.CrossRefGoogle Scholar
  5. Brakenhoff, G.J., and Visscher, K., 1993, Imaging modes for bilateral confocal scanning microscopy, J. Microsc. 171:17–26.CrossRefGoogle Scholar
  6. Denk, W., Strickler, J.H., and Webb, W.W., 1990, Two-photon laser scanning fluorescence microscopy, Science 248:73–76.PubMedCrossRefGoogle Scholar
  7. Pawley, J.B., 1994, The sources of noise in three-dimensional microscopical data sets. In: Three-Dimensional Confocal Microscopy: Volume Investigation of Biological Specimens (J. Stevens, ed.), University of Toronto, Academic Press. San Diego.Google Scholar
  8. Petráň, M., and Hadravsky, M., 1968, Tandem-scanning reflected light microscope, J. Opt. Soc, 58:661–664.CrossRefGoogle Scholar
  9. Sheppard, C.J.R., 1992, Signal-to-noise in confocal microscope systems, J. Microsc. 168:209–218.CrossRefGoogle Scholar
  10. Tsien, R.Y., and Harvotrinian, A.T., 1990, Practical design criteria for a dynamic ratio imaging system, Cell Calcium 11:93–109.PubMedCrossRefGoogle Scholar
  11. Tsien, R.Y., and Waggoner, A., 1989, Fluorophores for confocal microscopy: Photophysics and photochemistry. In: Handbook of Biological Confocal Microscopy (J.B. Pawley, ed.), Plenum Press, New York.Google Scholar
  12. van den Engh, G., and Farmer, C., 1992, Photo-bleaching and photon saturation in flow cytometry, Cytometry 13:669–677.PubMedCrossRefGoogle Scholar
  13. van der Voort, H.T.M., Brakenhoff, G.J., and Baarslag, M.W., 1989, Three-dimensional visualization methods for confocal microscopy, J. Microsc. 153:123–132.PubMedCrossRefGoogle Scholar
  14. Wells, K.S., Sandison, D.R., Strickler, J., and Webb, W.W., 1989, Quantitative fluorescence imaging with laser scanning confocal microscopy. In: Handbook of Biological Confocal Microscopy (J. Pawley, ed.), Plenum Press, London, pp. 27–39.Google Scholar
  15. White J.G., 1991, Confocal imaging system, U.S. Patent No. 5.032,720.Google Scholar
  16. Xiao, G.Q., and Kino, G.S., 1987, A real-time confocal scanning optical microscope, Proc. SPIE 809:107–113.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • G. J. Brakenhoff
    • 1
  • K. Visscher
    • 1
  1. 1.Section of Molecular Cytology, Institute for Molecular Cell Biology, BioCentrum AmsterdamUniversity of AmsterdamAmsterdamThe Netherlands

Personalised recommendations