migration assay - (Oct/19/2006 )

hey everyone,could somebody help me??????????
i'm developing a migration assay on PET membrane.
The hypotetic migrated cells, human dermal fybroblasts, have no stretch shape as normal
now....what's happend?it's a migration consequence?it's normal??

thanks everyone

greatings

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if they migrate they should be stretched; maybe its better to coat and try different coats; it depends on the aim of your study as the quality of adhesion triggers signaling processes;

as you try to test migration I suppose you analyze transgenic or pharmacological effects, maybe morphological changes are consequence of genetic or pharmacological manipulation...

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thanks a lot...my treatment doesn't cause morphologic changes on slides cultures, on the upper side of the pet membrane the cell are really nice but the problem is on the bottom...
I thought was a problem of fixation with metanol...
so the cell after migration have to be stretch????as in normal cultures???or could i use the results as good???

thanks in advances

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I mean when they migrate they are normally strechted; if they do not migrate can be stretched or can round up; I think it depends on culture condition, matrix and so on;

To make it more clear: "on the bottom" means you use a two chamber culture systems such as Transwell plates with wide pores (3/5/8 µm); and they migrate to the lower chamber ("bottom"); is upper and lower chamber of the same material (PET)? fixation with -20°C methanol is normally very rapid; I cannot exclude that it is the fixation method; I again recommend to check with coated plates (Collagen, polylysine etc)

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