Re: Measuring PSFs using water dipping objective

Posted by mmodel on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Measuring-PSFs-using-water-dipping-objective-tp7578494p7578497.html

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Hi Stephan,

You can try polyvinyl acetate. It is soluble in methanol (1% w/v) but not in water, and if you mix its solution with beads and dry it on a glass slide, some beads may get trapped in PVA and stay there even after you add water.

Mike

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Stephan Junek
Sent: Wednesday, June 13, 2012 12:35 PM
To: [hidden email]
Subject: Measuring PSFs using water dipping objective

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Dear all,

I am looking for a good protocol to measure PSFs on a confocal microscope
using water dipping objectives (objectives used for physiology, i.e. no
cover glass!).
I tried using fluorescent beads embedded in 5 % low melting point agarose,
but it appears that the beads are moving, probably due to slight swelling of
the agarose which is immersed in the water.
I would prefer not to use a cover glass, as this probably introduces
aberrations. Using beads dried on a glass surface is not ideal either due to
the reflection at the water/glass interface.

Thanks in advance for any suggestions!

Best,
Stephan.


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Dr. Stephan Junek
Max Planck Institute for Brain Research
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60528 Frankfurt am Main
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