Point Spread Function Software for Confocal Microscopy
- 詳細技術說明
- The point spread function (PSF) of an optical imaging system such as a microscope describes the optical characteristics of the system, including inherent diffraction effects, possible additional blurring imperfections, or aberrations. The PSF can be understood as the fundamental building block of the imaging process.
- *Abstract
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The resulting blurred image is the convolution, point by point, of the real object with the PSF of the system. PSF Lab is a software program that allows the calculation of the illumination PSF of a confocal microscope under various imaging conditions. The calculation of the electric field vectors and the associated light intensity distribution 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 to simulate an immersion oil/air (layer 1) objective that focuses 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.
While it is possible to measure the PSF of a microscope experimentally, it is difficult, and might have to be repeated for different experimental conditions. A good theoretical calculation of a PSF is very important as the quality of the PSF impacts directly on the quality of the deconvolved image. The quality of PSFs currently used in commercial imaging varies widely due to the complex underlying theory. The PSF Lab software represents the most complete software package for calculating PSFs available today and should greatly improve the quality of deconvolved images.
- *IP Issue Date
- None
- *IP Type
- Download
- *Principal Investigation
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Name: Michael Nasse, Assistant Scientist
Department:
- *Publications
- Nasse M. J., Woehl J. C. (2010). Realistic modeling of the illumination point spread function in confocalscanning optical microscopy. J. Opt. Soc. Am. A 27 (2): 295–302. Nasse, M.J., Woehl, J.C., and Huant S. (2007). High-resolution mapping of the three-dimensional pointspread function in the near-focus region of a confocal microscope. Applied Physics Letters 90 (3). Software Download Available At: http://onemolecule.chem.uwm.edu/software
- 申請號碼
- Non-Confidential Summary
- 國家/地區
- 美國
