SIM redux

Posted by mcammer on
URL: http://confocal-microscopy-list.275.s1.nabble.com/SIM-redux-tp7580938.html

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We have a practical question about how we know whether a SIM result is showing real structure or not.

We know from EM that the structure we are looking at is densely packed solid containing the protein of interest throughout, not hollow.  However, the SIM result appears hollow.  There is a possibility that the staining doesn't penetrate to the core, but we really don't know.

Has anybody see this sort of problem with SIM where a structure known to be solid is reconstructed with a hollow core?

Illustration at http://www.flickr.com/photos/mcammer/9696746798/ or https://nyumc.box.com/s/gyq3c3cw6p5aofb7gw84

We are using OMX Blaze with a 568 (or is it 561) nm laser.

Thank you.

_________________________________________
Michael Cammer, Assistant Research Scientist
Skirball Institute of Biomolecular Medicine
Lab: [phone]  Cell: [phone]