Improve Research Reproducibility A Bio-protocol resource

我的步骤是：肿瘤研磨和消化成单细胞（酶加的就是上述protocol推荐的），洗两遍（200g~400g离心力都用过，P...

Victor Chen Jul 15, 2019

Protocol: Isolation of Mouse Tumor-Infiltrating Leukocytes by Percoll Gradient Centrifugation

我的步骤是：肿瘤研磨和消化成单细胞（酶加的就是上述protocol推荐的），洗两遍（200g~400g离心力都用过，PBS和1640和DMEM培养基都用过），裂红后中和，离心后用40%的percoll重悬，用长的骨穿针在底部加80%percoll（40%与80%percoll的体积比例2:1和1:1都用过，50ml和15ml离心管都用过，觉得还是50ml的管效果比较好），离心20min以上（20min~35min时间都用过，刹车0~5都试过，转速从325g~800g，即转速从1400rpm~2000rpm都用过），以上条件结果全都是肿瘤细胞在最上层。中间一层是肿瘤细胞和免 Read more more

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冬刘 Answered Oct 11, 2021

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Yes, what I did before is the same as you. Some tumor cells are well separated in the lower layer, and some are in the upper layer. It is estimated that the concentration is inconsistent. But no matter what the form is, the middle layer is mixed and will not be completely separated. One thing is that when running streaming, different cell sizes can still be distinguished.

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