Sticky cells - (Mar/09/2006 )
I’ve been having a problem counting me cells.
After trypsinising (0.25% trypsin, 0.02% EDTA) the cells I spin them, wash them and try to count then but they clomp up in groups of 3 or more.
Anyone have an idea?
here you not gives detailes of problems which you face. We are also using J774 cell line before counting we trypsiniz its .
I thinks in your cases you trypsini9ze its for a long time thats why such type of problem you facing.
according to my experience before trypsininzing remove all media in 15ml cetrifuge tube and add approx that much amount of trypsin and EDTA solution its just cover the surface of cells and watch cairefully in microscope when you see appearence of crack in the monolayer of cells , tapps flask and add quckly media in the flask , now you try to count these cells...............but keep in mind dont keeps cells alone in trypsin solution for long time.
all the best.
I usually count them before spinning (well I take an aliquot out to count and spin the rest). I'm assuming you're using a haemocytometer for counting.
how about the G-force you seted in centrifuge. With lower speed, you, perhaps, will get better result.