Lesson plan

PSF Lab

PSF Lab is a software program that allows the calculation of the illumination point spread function (PSF) of a confocal microscope under various imaging conditions. The calculation of the electric field vectors is based on a rigorous, vectorial model that takes polarization effects in the near-focus region and high numerical aperture microscope objectives into account. The polarization of the input beam (assumed to be collimated and monochromatic) can be chosen freely (linear, circular, or elliptic). Furthermore, a constant or Gaussian shaped input beam intensity profile can be assumed. On its way from the objective to the focus, the illumination light passes through up to three stratified optical layers, which allows the simulation of an immersion oil/air (layer 1) objective that focusses light through a glass cover slip (layer 2) into the sample medium (layer 3). Each layer is characterized by its (constant) refractive index and thickness. PSF Lab can also simulate microscope objectives that are corrected for certain refractive indices and cover slip thicknesses (design parameters). Thus, any deviations from the ideal imaging conditions for which the objective was designed for are properly taken into account.…

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Article summary

PSF Lab is a software program that allows the calculation of the illumination point spread function (PSF) of a confocal microscope under various imaging conditions. The calculation of the electric field vectors is based on a rigorous, vectorial model that takes polarization effects in the near-focus region and high numerical aperture microscope objectives into account. The polarization of the input beam (assumed to be collimated and monochromatic) can be chosen freely (linear, circular, or elliptic). Furthermore, a constant or Gaussian shaped input beam intensity profile can be assumed. On its way from the objective to the focus, the illumination light passes through up to three stratified optical layers, which allows the simulation of an immersion oil/air (layer 1) objective that focusses light through a glass cover slip (layer 2) into the sample medium (layer 3). Each layer is characterized by its (constant) refractive index and thickness. PSF Lab can also simulate microscope objectives that are corrected for certain refractive indices and cover slip thicknesses (design parameters). Thus, any deviations from the ideal imaging conditions for which the objective was designed for are properly taken into account.…

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Discussion questions

  1. Summarise the article in your own words. Which single claim would you verify first, and why?
  2. Which claim in this article is most likely to be revised as the sources update? How would you check its current status?
  3. If two sources disagreed about one of these facts, what would you need in order to trust one over the other?

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Key claims and bibliography are generated from the same tracked claims and sources as the article page.