Just how many cells are lost when changing tubes? - (Sep/28/2007 )
Just how many cells are lost when changing tubes? This is what I was thinking when I poured at least 10ul of cells from one 50ml tube into a 15ml tube and lost all visable signs of any cells. I had nothing at the bottom of this tube after centrifuging it at 300g for 15 minutes! Please share any similar experiences and how to correct this problem. The 50ml tube was Sigmacoated. 
Are you sure you take the cells?? May be they were at the bottom and you just take medium in the 50 ml falcon.
I don’t think lot of cells are lost in the process of changing tubes.
ok when i use to establish cell lones, i resuspend 15 cm plate in 50ml and pick 10µl to seed 10cm plate.
About 25 to 50 clones shows up.
So i don't guess you'll gonna see any visible pllet. In same way, i think if you have let's say 100cells, i don't think you can do analysis on such few material
I don’t think lot of cells are lost in the process of changing tubes.
Hi and thank you for your responce. The mnc are clearly at the 1.075g/ml interface, and then with a Sigmacoated glass pipette, placed (suspended) at the bottom of a PP 15ml tube with about 10cc of total fluid (Hank's). Then this tube is spun down and THE CELLS ARE GONE.
About 25 to 50 clones shows up.
So i don't guess you'll gonna see any visible pllet. In same way, i think if you have let's say 100cells, i don't think you can do analysis on such few material
Hi Fred,
The 10ul of mnc I use are condensed and number approximately 3 - 5 million. You need at least 100 cells per ul for flow cytometry work. It just seem the more substrates the cells come in contact with the less of them you have.
Does anyone think that the problem is because cells stick to plastic when there isn't any serum or protein around? What about temperature? 