小中大
看到许多朋友的贴子,我心里真的感觉很沉重。我不多说了,只把我过去发表在中国生物论坛的贴子转过来,希望对需要的朋友有些帮助。当然,许多人不需要看。
ylcui 发表于 2012-07-05.02:48:21
DMSO的沸点很高,应该说摄氏121度不会沸腾。但是考虑到容器问题,和安全性问题,大家都不这么做。
Corning公司在他们的Protocol中推荐过滤除菌,但是注意,一定不能用普通的醋酸纤维素膜或PET膜的无菌滤器,不能耐受DMSO的腐蚀。应该用RC膜或聚四氟乙烯膜无菌滤器过滤除菌。Corning公司有专门的产品出售,14元一个。
ylcui发表于 2012-09-14.23:33:00
怎见得您买来的DMSO就是无菌的呢?请看看国外的一些Protocols吧。
科学实验不能够想当然,一定要参考文献,除非您做的是独创性的工作,需要建立新的试验方法。否则不按常规方法操作,试验的结果,很难为国外同行认可。
Coring公司的参考资料:(有关内容在第4页)
cuturl('http://www.corning.com/LifeSciences/pdf/t_cryoanimalcc.pdf')
以下论述摘自NUNC公司的一个小册子,Cryopreservation Manual,在cuturl('http://www.nuncbrand.com/')可以找到。
Glycerol and DMSO should be of reagent grade or better,
sterilized prior to use, and examined for undesirable properties.
Each lot should be examined for toxic properties by exposing
sensitive cells to concentrations previously used with success.
Glycerol may be sterilized by autoclaving for 15 minutes at
121°C and 15 psig. Glycerol should be protected from light
during storage. DMSO must be sterilized by filtration using a
0.2-micron nylon syringe filter or a Teflon PTFE* syringe filter
which has been pre-washed with alcohol and rinsed with
DMSO. Cryoprotective agents should be prepared in singleuse
volumes to minimize the risk of contamination and moisture
introduction with repeated use from one container.