Can we use very small beads in combination with averaging to obtain a PSF?
The averaging procedure can increase the signal of the accumulated bead as much as needed BUT it makes an alignement error which gets worse when the signal becomes poor. The alignement error effectively increases the bead size, but not in a way which can be predicted easily.
As soon as the bead signal is sufficient for alignment the SNR can be increased by averaging, but be careful if the signal is insufficient. Beads on the range of the 50nm seldom deliver enough signal. On very sensitive microscopes 110nm beads are good enough, but this is probably not the case on regular commercial microscopes.
Keywords: PSF beads averaging
Categories: Faq Deconvolution, Faq Microscopy, Huygens Faq, Imported Faqs
Platforms: Irix Linux Windows Mac
Related products: Hu Pro
