Posted by
Ian Dobbie on
URL: http://confocal-microscopy-list.275.s1.nabble.com/SIM-redux-tp7580938p7580946.html
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Peng Xi <
[hidden email]> writes:
> When we use SIM to image the particles, we found surprisingly that
> even with 200nm out of focus, the reconstructed image shows hollow
> structure, exactly as what you have seen. The hollow appears to be
> insensitive to the parameters of reconstruction process.
> Based on the image you provided, I guess it may be from the fact that
> your structure is slightly above the focal plane -- you may want to do a z
> series of SIM to verify it.
> You can download the original data from:
>
http://bme.pku.edu.cn/~xipeng/FND-SIM.rar Please pay special attention
> to the last two frames.
This data is pretty horrible to be honest. Scanning through the stack
there looks to be a squint PSF, or illumination. An orthogonal view
shows that the XZ PSF is bent. Further doing a FFT on the presented data
shows that there is no information beyond about 220 nm, so the image
isn't super resolution at all. I don't think that you can use this data
to show that a single molecule emitter produces a hollow above or below
focus.
Ian