Point Spread Function calculations for fluorescence microscopy
Project description
Psf is a Python library to calculate Point Spread Functions (PSF) for fluorescence microscopy.
This library is no longer actively developed.
- Authors:
Christoph Gohlke, Oliver Holub
- Organization:
Laboratory for Fluorescence Dynamics. University of California, Irvine
- License:
3-clause BSD
- Version:
2019.4.22
Requirements
Matplotlib 2.2 (optional for plotting)
A Python distutils compatible C compiler (build)
Revisions
- 2019.4.22
Fix setup requirements. Fix compiler warning.
- 2019.1.1
Update copyright year.
References
Electromagnetic diffraction in optical systems. II. Structure of the image field in an aplanatic system. B Richards and E Wolf. Proc R Soc Lond A, 253 (1274), 358-379, 1959.
Focal volume optics and experimental artifacts in confocal fluorescence correlation spectroscopy. S T Hess, W W Webb. Biophys J (83) 2300-17, 2002.
Electromagnetic description of image formation in confocal fluorescence microscopy. T D Viser, S H Wiersma. J Opt Soc Am A (11) 599-608, 1994.
Photon counting histogram: one-photon excitation. B Huang, T D Perroud, R N Zare. Chem Phys Chem (5), 1523-31, 2004. Supporting information: Calculation of the observation volume profile.
Gaussian approximations of fluorescence microscope point-spread function models. B Zhang, J Zerubia, J C Olivo-Marin. Appl. Optics (46) 1819-29, 2007.
The SVI-wiki on 3D microscopy, deconvolution, visualization and analysis. https://svi.nl/NyquistRate
Theory of Confocal Microscopy: Resolution and Contrast in Confocal Microscopy. http://www.olympusfluoview.com/theory/resolutionintro.html
Examples
>>> import psf >>> args = dict(shape=(32, 32), dims=(4, 4), ex_wavelen=488, em_wavelen=520, ... num_aperture=1.2, refr_index=1.333, ... pinhole_radius=0.55, pinhole_shape='round') >>> obsvol = psf.PSF(psf.GAUSSIAN | psf.CONFOCAL, **args) >>> print('%.5f, %.5f' % obsvol.sigma.ou) 2.58832, 1.37059 >>> obsvol = psf.PSF(psf.ISOTROPIC | psf.CONFOCAL, **args) >>> obsvol[0, :3] array([ 1. , 0.51071, 0.04397]) >>> # save the image plane to file >>> obsvol.slice(0).tofile('_test_slice.bin') >>> # save a full 3D PSF volume to file >>> obsvol.volume().tofile('_test_volume.bin')
Refer to the psf_example.py file in the source distribution for more.
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