thanks to the reply. some antigens will be damaged by PFA fixation and can not be retreieval. the shock frozen section with acetone is of quite bad tissue quality. therefore I am trying alcohol!
2009-05-16 TF 发件人： Geoff McAuliffe 发送时间： 2009-05-15 23:55:15 收件人： tifei 抄送： Histonet@lists.utsouthwestern.edu 主题： Re: [Histonet] 70% alcohol fixation of brain Greetings TF: Good luck freezing a brain that is full of alcohol! Have you checked the freezing point of alcohol? Why are you doing this? Immersing a whole brain in 70% alcohol, why?? 30% sucrose is a cryoprotectant so the brain is not full of holes from ice crystals.Alcohol defeats this purpose. Geoff TF wrote: > Hi, i dont want to use PFA for the brain fixation (rat). > Now I tried to perfuse the rat with saline, followed with 70% alcohol. Then I > do post-fixation at room temperature for 24 hours. > I also tried saline perfusion, then I directly put the whole brain into 70% > alcohol. > Is this fine? > > Also, for dehydration before cutting frozen sections on a cryostat/microtome, > should I use 95% alcohol? I am now using 30% sucrose~ > > 2009-05-15 > > > > TF > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcaul...@umdnj.edu **********************************************
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