Posted by
Kyle Michael Douglass on
URL: http://confocal-microscopy-list.275.s1.nabble.com/PSF-asymetry-tp7583681p7583687.html
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Hi Christophe and Professor Xi,
The images are of fluorescent beads, which should have many different
fluorescent labels attached to their surfaces with dipoles pointing in
many different directions. I don't think that any single dipolar
emission pattern should be obvious since the patterns from all the other
labels on the same sphere should average out, but I could be wrong about
this.
I've also seen this same aberration in images of 100 nm fluorescent
beads on a custom microscope with multiple NA >= 1.45, 100x oil
immersion objectives. No degree of alignment of the imaging pathway
seems to fully correct it, at least in my hands. In the end, I also
suspected coverslip tilt based on the Arimoto paper and this
dissertation from Stefan Hell's lab:
https://ediss.uni-goettingen.de/bitstream/handle/11858/00-1735-0000-000D-F0B1-E/berning.pdf?sequence=1Best,
Kyle
On 04/21/2015 03:01 PM, Peng Xi wrote:
--
Kyle M. Douglass, PhD
Post-doctoral researcher
The Laboratory of Experimental Biophysics
EPFL, Lausanne, Switzerland
http://kmdouglass.github.iohttp://leb.epfl.ch