Posted by
Zdenek Svindrych-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/darkfield-references-tp7583371p7583373.html
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Hi Jeff,
I bet you can see 50 nm particles, especially when 'dry mounted'. The best
way is to try it yourself. Most today's (fluorescent) microscopes do not
have immersion condensers, but you may get a pretty good idea if you use e.
g. Olympus IX70 / IX80 microscope 'PH3' aperture for illumination and 10x-20
x objective (NA < 0.4).
To detect weakly scattering/refracting objects you need 1. clean sample, 2.
clean condenser. Quantitative treatment is always challenging. You may try
to read on Mie theory, Raighley scattering and the like...
Good luck!
zdenek
--
Zdenek Svindrych, Ph.D.
W.M. Keck Center for Cellular Imaging (PLSB 003)
University of Virginia, Charlottesville, USA
http://www.kcci.virginia.edu/workshop/index.php---------- Původní zpráva ----------
Od: Jeff Spector <
[hidden email]>
Komu:
[hidden email]
Datum: 30. 1. 2015 18:06:36
Předmět: darkfield references
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Greetings,
Can someone please point me to some references involving the theory
behind darkfield microscopy? I understand the basic idea, but all I can
find are different iterations of the basic idea that you block most of the
light and only image scattered light. I'd like to learn a bit more about
technical aspects of darkfield, i.e. what is the smallest object you can
observe? What role do illumination power and camera exposure play in the
quality of the final image. What role does specimen thickness/size play in
the final image and can you discern objects of different size
etc...
Any help would be greatly appreciated. Perhaps I simply need to read up on
scattering theory?
thanks..
-jeff"