Inconsistant Matrigel Invasion Assay - (Jun/01/2010 )
I have been attempting to optimise a matrigel invasion assay looking for differences in the invasive qualities of un-irradiated breast epithelial cells and irradiated (and potentially transformed) breast epithelial cells. I have been using the Matrigel coated Inserts from BD Biosciences and have had some success with the assay however my results appear to be very inconsistent. Using the same cells, chemoattractants and staining techniques over different experiments I do not get a consistent number of cells invading. Also my cells sometimes appear to invade uniformally over the insert surface and sometimes cells are only present around the edges of the insert and hardly any in the middle. Has anybody else had this problem and if so how did they get around it? Also does anyone have any good tips for when using this assay kit in general?
I have had the same issues and refuse to use the assay. It's a shame because it has the potential to be a very valuable tool for investigating motility and invasion, but the results are ridiculously inconsistent. Other lab mates of mine will use the assay and will defend the results, but I don't trust the results at all. There are too many variables that can skew the results. I understand that if you do enough replicates it should average out, but I just don't see that and don't have confidence in the technique.