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Quantitative Fluorescence Confocal Laser Scanning Microscopy (CLSM)

  • David R. Sandison
  • Rebecca M. Williams
  • K. Sam Wells
  • James Strickler
  • Watt W. Webb
Chapter

Abstract

The confocal imaging geometry provides a dramatic optical advantage for fluorescence microscopy by discriminating against out-of-focus background with minimal loss of image-forming signal. Significant enhancement of both axial and lateral imaging resolution is also available but only with substantial signal loss. Because of these optical advantages, the confocal laser scanning microscope (CLSM) can clearly image thin optical sections from within thick fluorescence-labeled living specimens. A stack of optical sections is easily combined to reveal three-dimensional (3D) fluorescent marker distributions with diffraction-limited spatial resolution. When bright stable fluorophores are available, cellular dynamics can be measured by recording a time series of CLSM images.

Keywords

Objective Lens Shot Noise Spherical Aberration Axial Resolution Chromatic Aberration 
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

  • David R. Sandison
    • 1
  • Rebecca M. Williams
    • 1
  • K. Sam Wells
    • 1
  • James Strickler
    • 1
  • Watt W. Webb
    • 1
  1. 1.Developmental Resource for Biophysical Imaging and Optoelectronics, Applied and Engineering PhysicsCornell UniversityIthacaUSA

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