aria II分选问题

seawidesky

Aria II分选问题，分选的样本细胞如果有很多碎片，分选的过程中阈值设置最小，但是分选后的目的细胞群上机复检的时候发现还是有很多碎片，这个有什么方法可以避免吗？

niwanmao

参考一下文献中的方法，在分选前去除：
Large debris and cell clusters were removed from the cell suspension by serial filtration through pre-wetted 100 μm (Falcon 352360; BD Biosciences, San Jose, CA) and then 40 μm (Falcon 352340; BD Biosciences) cell strainers into 50 ml Falcon tubes on ice. Small cellular debris was reduced by density centrifugation through a three-density step gradient of Percoll (P1644; Sigma, St. Louis, MO). One milliliter of each solution (high density solution: 3.426 ml Hibernate A + 824.5 μl Percoll + 97.8 μl of 1M NaCl; medium density solution: 3.600 ml Hibernate A + 650.5 μl Percoll + 76.5 μl of 1M NaCl; low density solution: 3.770 ml Hibernate A + 480.3 μl Percoll + 59.5 μl of 1M NaCl) was carefully layered in a 15 ml Falcon tube, with the highest density solution on the bottom. The filtered cell suspension was applied to the top of this gradient and centrifuged at 430×g for 3 minutes; longer centrifugation times did not produce the desired separation. The cloudy top layer (~2 ml) containing debris was removed and discarded. Cells in the remaining layers were pelleted by centrifugation at 550×g for 5 min.
（来源：http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3221768/）