How to check the size of a bead as it appears in the image?
My beads appear to be much larger than their actual size, what is wrong?
Beads with a size close to the width of the PSF half intensity width (HIW, the width of the PSF at 50% of the peak intensity) show up in the image as spots which are slightly wider than the PSF. <br>
To compare a bead image to a theoretical PSF proceed as follows: <br>
-Open an operation window on the bead or averaged bead image <br>
-Choose a free destination image <br>
-Click the PSF button and set Dimensions to Parent <br>
-Click Run to generate a theoretical PSF of which the size matches the size of the bead image
You can now compare the two images by opening a Slicer window on the PSF destination image
If you find that the beads are much larger than the PSF you might be looking at bead clusters. In that case you might find be able to find weak single bead images in the background. Because the clusters are of unknown size you can't use them for PSF measurement. <br>
Alternatively there might be large beads in the sample. If these have an unknown size, or if their size is larger than the HIW, best not use them for PSF measurement.
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Keywords: PSF measurement bead HIW<br>
Categories: Faq Deconvolution, Faq Microscopy, Huygens Faq, Imported Faqs<br>
Platforms: Irix Linux Windows Mac AIX<br>
Related products: Hu Ess Hu Pro<br>
