Posted by
Carol Heckman on
URL: http://confocal-microscopy-list.275.s1.nabble.com/New-building-options-feedback-request-from-facilities-NOT-located-on-ground-floor-basement-tp3889815p3893371.html
Ralf-
There is a reference to enhanced phase contrast in a recent paper. This might help, as the phase retardation introduced by water may be quite small. You can find it (open source) by searching Vasiliev and "enhanced phase contrast".
Carol
Center for Microscopy & Microanalysis
Bowling Green State University
________________________________________
From: Confocal Microscopy List [
[hidden email]] On Behalf Of Ralf Zenke [
[hidden email]]
Sent: Monday, October 26, 2009 6:12 AM
To:
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Subject: Quantification of phase contrast images
Dear list,
For pre-observation of samples for electron microscopy we need to measure the thickness of amorphous ice (~500nm) on thin (~10nm) carbon meshes. We would like to use phase contrast (I think DIC could not be quantified in that experiment because of the homogeneous thickness of the ice layer).
Is it possible to get reliable thickness values (accuracy of ca. 50nm) out of the gray values of the camera? Reference images without ice layers can easily be aquired. Are there any effects that might corrupt the measurements? I think it's at least neccessary to have a narrowband light source.
Thanks to anyone who can give input on this!
Nice regards,
Ralf
Ralf Zenke
Max Planck Institute of Biochemistry
Core Facility
Am Klopferspitz 18
DE-82152 Martinsried near Munich
GERMANY
Phone: (+49) (89) 8578 3798
Fax: (+49) (89) 8578 2847
www.biochem.mpg.de<
http://www.biochem.mpg.de>