Measurement of point-spread function (PSF) for confocal fluorescence microscopy

In this paper, we describe size-effect of fluorescent microsphere for measuring point-spread function (PSF) in confocal fluorescence microscopy. We present the numerical results for the practically available microsphere size range for measuring PSF, and demonstrate with experiment. Also, the effective PSF is restored with deconvolution technique within an acceptable error. Also the size-effect for measuring phase modulated PSF, which has two vicinal peaks, is described. The numerical and experimental result is also presented.