Advertisement

Confocal Microscopy with Transmitted Light

  • A. E. Dixon
  • Carol Cogswell

Abstract

Although confocal reflection (brightfîeld) and fluorescence microscopy have found widespread use in biology, scanning transmission microscopy has been explored and implemented only infrequently, and a single-pass scanning confocal transmission microscope is still not available as a commercial product. This might indeed be considered surprising, since biologists have long known the advantages of observing an enormous variety of specimens (e.g., stained or unstained, preserved or living) using conventional light microscopes with their wide range of transmission modes such as brightfîeld, phase-contrast, or differential interference contrast (DIC). The fact that confocal transmission modes are not generally available is partly due to the fact that most biological preparations have a complex mixture of optical properties that create regions of varying refractive index (η) and absorption. These can introduce sufficient aberration and deflection into the transmitted wavefront to make it very difficult to keep the transmitted beam aligned with a standard pinhole detector as it scans over the specimen.

Keywords

Microscope Objective Transmission Image Transmitted Beam Reflection Image Wollaston Prism 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Allen, R.D., David, G.B., and Nomarski, G., 1969, The Zeiss-Nomarski differential interference equipment for transmitted-light microscopy, Z. Wiss. Mikrosk. 69:193–221.PubMedGoogle Scholar
  2. Art, J.J., Goodman, M.B., and Schwartz, E.A., 1991, Simultaneous fluorescent and transmission laser scanning confocal microscopy, Biophys. J. 59:155a.Google Scholar
  3. Brakenhoff, G.J., 1979, Imaging modes in confocal scanning light microscopy (CSLM), J. Microsc. 117:232–242.Google Scholar
  4. Cogswell, C.J., and O’Byrne, J.W., 1992, A high resolution confocal transmission microscope: I. System design, Proc. SPIE 1660:503–511.CrossRefGoogle Scholar
  5. Cogswell, C.J., and Sheppard, C.J.R., 1989, Imaging using confocal brightfield techniques, Inst. Phys. Conf. Ser. 98:633–638.Google Scholar
  6. 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. Microsc. 165:81–101.CrossRefGoogle Scholar
  7. Dixon, A.E., Damaskinos, S., and Atkinson, M.R., 1991a, Transmission and double-reflection scanning stage confocal microscope, Scanning 13:299–306.CrossRefGoogle Scholar
  8. Dixon, A.E., Damaskinos, S., and Atkinson, M.R., 1991b, A scanning confocal microscope for transmission and reflection imaging, Nature 351:551–553.CrossRefGoogle Scholar
  9. Goldstein, S., 1989, A no-moving-parts video rate laser beam scanning type 2 confocal reflected/transmission microscope, J. Microsc. 153:RP1-RP2.CrossRefGoogle Scholar
  10. Goldstein, S., Hubin, T., Rosenthal, S., and Washburn, C., 1990, A confocal video rate laser beam scanning reflected light microscope with no moving parts, J. Microsc. 157:29–38.PubMedCrossRefGoogle Scholar
  11. O’Byrne, J.W., and Cogswell, C.J., 1992, A high resolution confocal transmission microscope: II. Determining image position and correcting aberrations, Proc. SPIE 1660:12–520.Google Scholar
  12. Sheppard, C.J.R., and Wilson, T., 1980, Multiple traversing of the object in the scanning microscope, Opt. Acta 27:611–624.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • A. E. Dixon
    • 1
  • Carol Cogswell
    • 2
  1. 1.Department of PhysicsUniversity of WaterlooWaterlooCanada
  2. 2.Physical Optics Department, School of PhysicsUniversity of SydneySydneyAustralia

Personalised recommendations