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This analysis only concerns microscopes equipped with a motorized Z stage.
The point spread function (PSF) determines the spreading behavior of light by a point-like object through an imaging system. The full width at half maximum (FWHM) of the PSF along X, Y and Z is a parameter commonly used to describe the system resolution. Fitting the PSF with a mathematical function (Gaussian, Lorentzian, Sech²) allows the determination of the FWHM.
The “point spread function” analysis provides information, in 3D, on how light spreads from a point-like object, as well as quantitative parameters such as the lateral and axial FWHM of the PSF and the signal-to-noise ratio (SNR) in the image.
The “point spread function” analysis is associated with any “point-like object” (see Figure 1).
Figure 1: Example image of a “point-like object”, fulfilling the acquisition recommendations