Theory of confocal fluorescence imaging in the programmable array microscope (PAM)

  • Verveer P
  • Hanley Q
  • Verbeek P
 et al. 
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Abstract

The programmable array microscope (PAM) uses a spatial light modulator (SLM) to generate an arbitrary pattern of conjugate illumination and detection elements. The SLM dissects the fluorescent light imaged by the objective into a focal conjugate image, Ic, formed by the ‘in-focus’ light, and a nonconjugate image, Inc, formed by the ‘out-of-focus’ light.We discuss two different schemes for confocal imaging using the PAM. In the first, a grid of points is shifted to scan the complete image. The second, faster approach, uses a short tiled pseudorandom sequence of two-dimensional patterns. In the first case, Ic is analogous to a confocal image and Inc to a conventional image minus Ic. In the second case Ic and Inc are the sum and the difference, respectively, of a conventional and a confocal image. The pseudorandom sequence approach requires post-processing to retrieve the confocal part, but generates significantly higher signal levels for an equivalent integration time.

Author-supplied keywords

  • Confocal microscope
  • Digital micromirror device (DMD)
  • Optical sectioning
  • Point-spread- function
  • Pseudorandom sequences
  • Spatial light modulator
  • Tandem scanning microscopy

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Authors

  • P. J. Verveer

  • Q. S. Hanley

  • P. W. Verbeek

  • L. J. Van Vliet

  • T. M. Jovin

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