I have done just what you are talking about using Aclar. It is a plastic embedding film (purchased from EMS). It works great for us.
Margaret E. Bisher Electron Microscopy & Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 [email protected] On 1/27/10 3:39 PM, "Sherwood, Margaret" <[email protected]> wrote: > Nick, > > I am assuming that your 3D cells only grow on plastic. They make plastic > cover > slips which, if your cells attach to them and grow, might make the embedding > much easier. You would follow the same method as stated, but then you could > invert the coverslips on a beem capsule and separate the coverslip from > capsule > with liquid nitrogen. However, I have never done it with plastic coverslips > (only glass), so not sure if they would easily separate from capsule with > liquid > nitrogen. If anyone else has done so, please weigh in. > > Peggy > > -----Original Message----- > From: [email protected] > [mailto:[email protected]] On Behalf Of Geoff > McAuliffe > Sent: Wednesday, January 27, 2010 3:20 PM > To: Nicholas David Evans > Cc: [email protected] > Subject: Re: [HISTONET] embedding cell cultures > > Hi Nick: > > You can use the Epon substitutes such as EmBed 812. Fix, osmicate, > dehydrate as usual, but omit the proplylene oxide as it will react with > the plastic dish. Epon substitutes will mix with ethanol. I used 2:1 > then 1:1 then 1:2 ratios of absolute ethanol to "Epon" with catalyst > added with agitation. Then several changes of pure Epon and polymerize. > Yes, you will have to saw away the plastic, if you try to section the > Epon-plastic combo the two will separate. > How much easier do you want it to be? > > Geoff > > Nicholas David Evans wrote: >> Dear all, >> >> I was hoping someone might be able to offer me some advice on embedding and > sectioning cell cultures. >> >> In short we are growing cells which form 3D dome-like structures on tissue > culture plastic. Does anyone have any experience or advice to offer on > embedding > the cultures in situ before sectioning? I have seen various methods in the > literature, which often use Epon to embed the material followed by sawing away > the plastic, but if anyone can offer some tips on other possible (easier) ways > of doing it, or can refer me to some useful literature, I'd be very grateful. >> >> I would like to have simple 10um sections at 90 degrees to the substrate, > which I can use for IHC. >> >> Best wishes >> Nick >> >> _______________________________________________ >> Histonet mailing list >> [email protected] >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
