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closeisotropic 3D morphometry of H&E is a waste of time
Posted by gmcnamara on 17 Dec 2012 at 18:51 GMT
The Nandakumar et al 2012 isotropic 3D morphometry of H&E is a waste of time. H&E is not - and never will be - quantitative.
They are mis-using the term "optical density". Compare the definition of O.D. in freshman chemistry to this paper.
The paper has (as is now usual) inadequate methods. I had to go to the ref 28 to discover they are claiming 350 nm resolution, and to the supplement (of ref 28) to diiscover they are using a 100x/1.3NA objective lens. which should provide 230 nm resolution (Airy equation at 500 nm). No mention of condenser NA, total magnification, and using a color CCD camera, so "resolution" might just be pixel size. For comparison, confocal microscopy would be 230x230x~700 nm and a 3D-SIM fluorescence nanoscope would be 115x115x~350 nm -- and with the right fluorophores would be multiparameter and quantitative.
The end of the discussion mentions the possibility of "fluorescence Cell-CT" (essentially free marketing for some future potential VisionGate product).