Measuring beads - apart from the bead size what else is important?
I understand the importance of the small size and the way Huygens calculates the psf from the bead size and the advantage of the brighter signal from the bigger beads.
The averaging procedure can increase the signal of the accumulated bead as much as needed. BUT it makes an alignment error which is worse when signal is poor. The alignment 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 if it is insufficient you're sunk. I've never seen enough signal from 50nm beads. On *very* sensitive microscopes 110nm beads are good enough, but I doubt whether they are safe to use on regular commercial microscopes.
For more information you can have a look at DeconvolvingBeads
Keywords: bead size averiaging
Categories: Faq Deconvolution, Huygens Faq, Imported Faqs
Platforms: Irix Linux
Related products: Hu Pro
