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Hello Jens,

we use poly-L-lysine (MW 300000) to get cells growing in suspension on
coverslips. After half an hour, they are usually attached enough to proceed. We
didn't realy test for the minimal time though.
Is it essential that the cells are in a cuvette? That's probably for
electroporation? Can't you use a transfection method where the cells are on the
coverslip from the beginning?
Agarose might give you trouble since only a minority of the cells will be very
close to the coverslip, reducing the resolution.

Good luck,
Steffen

--- Jens Rietdorf <[log in to unmask]> schrieb: > Search the CONFOCAL
archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear listers,
>
> a client of our facilities wants to perform the following experiment:
>
> -transfect fluorescently labeled dna using different electroporation
> methods.
> -observe distribution of the dna by microscopy and follow up dna entry
> into cell nuclei.
>
> The transfection is taking place in a cuvette, therefor the main
> technical problem is to get the cells as soon as possible after the
> transfection into a microscopy compatible form, i.e onto a coverslip.
>
> Any of you did a similar experiment? How to proceed after the
> transfection? Use Cytospin? Low melting agarose embedding? Other
> strategy?
>
> Thanks for your comments,
> Jens
>
>
> --
>
> Dr. Jens Rietdorf
> EMBL, Meyerhofstr.1,
> D-69117 Heidelberg,
> Cell Biology/Biophysics Program,
> Advanced Light Microscopy Facility
> Phone ++49(0)6221 387-467 FAX-306
> http://www.embl-heidelberg.de/~rietdorf/index.html
>
http://www.embl-heidelberg.de/ExternalInfo/almf/htdocs/almf_website/index.html

=====
Steffen Dietzel, Dr. rer. nat.
Department Bioligie II - Humangenetik
Ludwig-Maximilians-Universität München
Richard-Wagner-Str. 10, 80333 München, Germany
phone: +49/89/2180-6728
fax:   +49/89/2180-6719
e-mail: [log in to unmask]  Web:  http://www.dietzellab.de

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