How to Fix Thin Layers of Matrigel? - ~0.5 mm layers of Matrigel break apart with formalin! (Sep/18/2008 )
I've been mixing cardiac myocytes with liquid BD Matrigel (basement membrane matrix), and plating it on glass-bottomed dishes. The gel hardens into thin layers (~0.5 mm) and supports cell growth for weeks without any breakdown of the gel integrity.
However, when I try to fix cells within the gel for confocal imaging, the gel layers often flake off and break apart. I've been using 10% formalin (4% paraformaldehyde) for 15 minutes. I've dropped it down to 2.5% formalin for 10 minutes, but it doesn't help much. (I've been able to get by for a while just by seeding tons of replicates, and some gel layers do hold on - though just barely - during the staining process.)
So does anybody know of a better way to fix cells in Matrigel?
we coat our plates with matrigel before adding cells. And when we fix, we do not have a problem.
try diluting matrigel and then mixing cells and plating them.