h***a
发帖数: 145 | 1 可以用trypsin吗？我们用了0.25%trypsin处理6个月的小鼠的hypothalamus 20mins at
37C,怎么感觉完全没消化啊。
用pasteur pipets 吹打的时候完全吸不进去。。崩溃。
高手指点一下吧。多谢啦！ |
h*******o
发帖数: 4884 | 2 It is a lot harder to culture adult neurons than embryonic neurons.
Our lab has tried for quite a while with no success.
If you are interested, take a look at Greg Brewer's Nature Protocol paper.
Good Luck |
h***a
发帖数: 145 | 3 怪我没说清楚， 我们不是要culture neurons, 只是要看hypothalamus里面的
inflammation,所以要得到的是all live cells，然后直接染抗体，上flow cytometer.
刚刚勉强用2ml pipets 吹打下来了，但是感觉dirt好多哦，在hemacytomer看的时候全
是些奇形怪状的东西。 |
h*******o
发帖数: 4884 | 4 That is the problem for both culture and your study
For adult tissue, it is hard to dissociate without damaging the cells. Most
of the stuff you get is actually chunks of tissue debris.
cytometer.
【在 h***a 的大作中提到】
: 怪我没说清楚， 我们不是要culture neurons, 只是要看hypothalamus里面的
: inflammation,所以要得到的是all live cells，然后直接染抗体，上flow cytometer.
: 刚刚勉强用2ml pipets 吹打下来了，但是感觉dirt好多哦，在hemacytomer看的时候全
: 是些奇形怪状的东西。
|
h***a
发帖数: 145 | 5 I agree. What I see under microscope is actually dirt and a few live cells.
Most
【在 h*******o 的大作中提到】
: That is the problem for both culture and your study
: For adult tissue, it is hard to dissociate without damaging the cells. Most
: of the stuff you get is actually chunks of tissue debris.
:
: cytometer.
|
