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Coated plates for fluorescence imaging of baculovirus-infectedinsect cells?

Coated plates for fluorescence imaging of baculovirus-infectedinsect cells? - Protocols and Methods Forum

Coated plates for fluorescence imaging of baculovirus-infectedinsect cells? - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 04-01-2008, 11:37 PM
Mike Autry
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Default Coated plates for fluorescence imaging of baculovirus-infectedinsect cells?



Methods:

Which coating is best to keep baculovirus-infected insect cells adhered to microscope plates: poly-lysine, integrin, laminin, etc? Any recommendationson plate type and vendor availability?

I have a major problem doing FRET microscopy on Sf21 insect cells infected with recombinant AcNPV baculoviruses: following infection, the Sf21 cells detach from the glass microscope plate and roll around, especially when I change filter cubes (donor, acceptor, FRET dichroics) during photobleaching experiments.

I am expressing ER membrane proteins tagged with cyan and yellow fluorescent proteins (CFP, YFP).

Thanks,

J. Michael Autry, PhD
Research Associate, University of Minnesota
Minnesota Muscle Laboratory, David D. Thomas, PI
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Old 04-02-2008, 05:22 AM
DK
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Default Coated plates for fluorescence imaging of baculovirus-infectedinsect cells?

In article <[Only registered users see links. ].net >, Mike Autry <[Only registered users see links. ].edu> wrote:

No experience with Sf21 cells in terms of trying to keep them attached but
in all difficult cases CellTak (available from BD Bioscience) was far superior
to everything you list. You can coat yourself - involves adding solution, waiting
and rinsing.

DK
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