Yes this is a very common approach... we experience difficulty partly because we are cutting thin sections on large samples from wounded tissue... the wounded tissue is a bit more fragile; also, if the section is large, it is very hard to use the small brush to hold it during cutting...
in the sense of brushing some water or PB (very tiny amount), i would like to ask the question that how can sections stick to slides? Will this water membrane (after you have mounted sections, it should be absorbed by the section) causes sections to peel off during IHC or other staining procedures? Is the warm temperature of a slide or the post-mounting dry procedure necessary for a nice adherence of sections? If we really know that, we may have the chance to reverse some "mis-mounted" sections to be firmly adhered to slides....? I dont believe that a tiny amount of salt between sections and slides can be a problem. We normally mount 20-40 um brain sections (floating sections prepared from microtome) in PB onto slides and dry it up. This approach never loses a single sections...even if you boiled it under 95 degree for half an hour. Can anyone tell the difference between this kind of mounting and cryostat mounting? Is OCT causing the peeling off of cryostat sections? I dont know, I hope someone can comment on these questions. 2009-04-29 TF 发件人： ooi.ting.huay 发送时间： 2009-04-29 13:15:46 收件人： tifei 抄送： Dearolf, Jennifer; histonet@lists.utsouthwestern.edu; histonet-bounces 主题： Re: [Histonet] Rolling sections Normally, we will use brush to hold the section after we have sectioned one third of the sample. After that, we will continue section the sample untill the end. If the sample is rolled up, we use the brush to smoothen it. Then quickly flip the glass slide to fish the sample. As the glass slide is keep in room temperature (warm), when put inside the cryostat machine (inside is cold), the section will just stick to the glass slide. This works quite well in our lab. Hope this can help... Regards, Ooi "TF" <ti...@foxmail.com> Sent by: histonet-boun...@lists.utsouthwestern.edu 04/29/09 11:00 AM Please respond to ti...@foxmail.com To"Dearolf, Jennifer" <dear...@hendrix.edu>, "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> cc SubjectRe: [Histonet] Rolling sections I also have problems in mounting brain sections...10um-30um in Leica Cryostat....hope anyone can share their experience? sometimes we use a brush to paint a mini drop of PB/water on to the slide, and then mount the sections. it is very flat then. 2009-04-29 TF 发件人： Dearolf, Jennifer 发送时间： 2009-04-29 06:56:50 收件人： histonet@lists.utsouthwestern.edu 抄送： 主题： [Histonet] Rolling sections Greetings, Histonetters! First, I wanted to thank all of you that responded to my e-mail a few years back about freezing small pieces of muscle tissue. We have found a method that works for us, and if anyone is interested, I would be happy to share. It still involves the wonderfully explosive isopentane, but it allows us to freeze fetal guinea pig muscle without artifact. I am writing today to ask a question about cutting frozen sections with a cryostat. We are having problems with the sections rolling once they come off the knife and before we can get them on a slide. We have a Microm 505E cryostat, and we cut our OCT mounted specimens at around -25 degrees C. We use Accuedge high profile blades, cut sections between 8 and 12 microns thick, and use a brush to pull the sections off. But, when we remove the brush, the sections roll up. Sometimes, they just arc up and other times they completely roll into a jellyroll. I have tried putting 70% EtOH in a beaker in the cryostat. This method was suggested to us by a vendor, but it doesn't seem to work consistently. We can also flatten the sections with a brush, but unless we are really quick, the sections roll up before we can get them on the slide. It makes it difficult to get serial sections. Any advice would be appreciated. Thanks again for all your help so far. Sincerely, Jenn Jennifer Dearolf, Ph.D. Associate Professor Biology Department Hendrix College 1600 Washington Ave. Conway, AR 72032 (501) 450-4530 (office) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________ Confidential information. Unauthorized use or disclosure prohibited. Refer http://www.singhealth.com.sg/ContactUs/#Disclaimer
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