I learned to freeze tissue by taking a beaker and filling it with 2-methylbutane and then placing that beaker in a liquid nitrogen bath. You then take place your cryomold, hold it using some long forceps, and place it on the top of the 2-methylbutane. It will freeze the entire thing in about 10 seconds. Once it's frozen, you dip the entire mold into the 2-methylbutane. Seems to work well for me.
Adam On Wed, Feb 24, 2010 at 4:13 PM, Andrea T. Hooper < [email protected]> wrote: > I have used the "Gayle Callis" method of freezing tissue for histological > applications for years with great success :) The tissues look fantastic. > > Essentially float out a metal pan (like the ones you sterilize surgical > tools in) on a liquid nitrogen bath (in a styrofoam container or large > dewar). Place fresh or cryoprotected tissue into cryomold (I like the Tissue > Tek ones mainly) and add sufficient OCT. Place mold onto metal tray and > close lid of dewar or styrofoam container. Let freeze. When frozen wrap in > foil and place into bitran bag, place into -80 deg C for long-term storage. > > If snap freezing tissue for RNA/protein extraction I use the classic > isopentane cooling method. > > Andrea T. Hooper > > > --- On Wed, 2/24/10, Laurie Colbert <[email protected]> > wrote: > > > From: Laurie Colbert <[email protected]> > Subject: [Histonet] Snap Freezing Tissue > To: [email protected] > Cc: "Steve Pike" <[email protected]> > Date: Wednesday, February 24, 2010, 9:54 PM > > > I was wondering how others are snap freezing tissue? We have been using > a histobath (a refrigerator device that cools isopentane for freezing > tissue), but it has died. I would love to have another Histobath, but > they are no longer carried by the vendor that I originally bought mine > from. Does anyone know where I can get either a new or refurbished > Histobath (vendor calls are welcome)? I'm also interested in other > freezing techniques. > > > > Laurie Colbert > > Huntington Hospital > > Pasadena, CA > > (626) 397-8620 > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > _______________________________________________ > Histonet mailing list > [email protected] > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
