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

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Published
1998-03
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  • http://onlinelibrary.wiley.com/termsAndConditions#vor
DOI
  • 10.1046/j.1365-2818.1998.00336.x
Publisher
Wiley

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<jats:p>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, <jats:italic>I</jats:italic><jats:sub>c</jats:sub>, formed by the ‘in‐focus’ light, and a nonconjugate image, <jats:italic>I</jats:italic><jats:sub>nc</jats:sub>, 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, <jats:italic>I</jats:italic><jats:sub>c</jats:sub> is analogous to a confocal image and <jats:italic>I</jats:italic><jats:sub>nc</jats:sub> to a conventional image minus <jats:italic>I</jats:italic><jats:sub>c</jats:sub>. In the second case <jats:italic>I</jats:italic><jats:sub>c</jats:sub> and <jats:italic>I</jats:italic><jats:sub>nc</jats:sub> 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.</jats:p>

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