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Fixing for Cell Migraton

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#1 PatrickNg



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Posted 02 February 2010 - 05:13 PM

Hello everyone,

I cant seem to find any other relevant section for my question, so I thought I'd post it in the Cell Biology Section.

I am currently conducting a cell migration assay using Mouse Embryonic Fibroblasts (MEFs) migrating through BD Falcon Cell Culture Inserts.

Post migration, I have the migrated cells
- Fixed with 4% PFA
- Stained with toludine blue

To view the migration of cells I am then to cut out the membrane and to fix them onto microscope slides.
However, when I search for possible mounting medium I come back with a handful of possible recipes.

I attempted mounting the membrane with 70% Glycerol in PBS, however when the membrane makes contact with the mounting media, some of the toludine blue seems to have reacted and dissapted from the membrane to the mounting medium.
I am unsure as to whether these are excess toludine blue (that are not cell stained) or if they were cell stained.

Does anyone know what sort of mounting media I can use that does not react with toludine blue causing me to lose some dye?

Thanks in advance


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