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Hi,
	Have you tried to activate your coverslip by putting it in a 
plasma for 30 sec to 1 min? When we do this, we find that the 
polylysine coating works much better than when the coverslips are 
only acid cleaned. Your friendly neighborhood EM lab should be able 
to provide a plasma for you to try.

	Hope it helps,
		Tobias

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>
>Dear all,
>we are trying to image fixed cell spheroids (>100um diameter) on a 
>leica sp5 inverted confocal microscope. The high resolution 
>objectives we use have short but sufficient working distance to 
>analyse the spheres if they attached to the  coverslip.  However 
>standard coatings as polilysine are not efficient in immobilizing 
>them (even if they can initially adhere we loose them extensively 
>during staining). We tried  also particular gels as Q gel but we 
>were not able to find a reproducible protocol to have them clearly 
>attached to the substrate  instead of being simply embedded in the 
>gel.
>I know that new high NA water-immersion long working-distance 
>objectives could provide the simplest solution but unfortunately 
>their cost is quite limiting.
>thanks for all the answers
>Mario
>
>--
>---PLEASE Note the change in telephone number---
>
>--
>Mario Faretta
>Department of Experimental Oncology
>European Institute of Oncology
>c/o IFOM-IEO Campus for Oncogenomics
>via Adamello 16 20139 Milan Italy
>Phone: ++39-0294375027
>email: [log in to unmask]
>http://www.ifom-ieo-campus.it


-- 
       _      ____          __   ____  
      /  \   /          / \    /   \ \       Tobias I. Baskin
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    /_ /   __      /__ \   \       \__    Biology Department
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  /      /          /         \   \       \	      University of 
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