Bad morphology of frozen cell
I was asked to prepare frozen cell blocks. The procedure is shown as below.
Could anybody give me suggestion to improve the morphology? Thanks. Wendy
1. Remove cells from culture flask. If antigen is internal, trypsin can
be used. If antigen is a cell surface epitope, remove cells from the
culture with a rubber policeman.
2. Using 50 ml polypropylene centrifuge tubes, gently centrifuge the cells
and pour off the culture medium. Wash cells with tissue culture medium,
centrifuge again, and pour off medium.
3. Add 0.5 ml of Tissue-Tek O.C.T. Compound to the cells in the tube
(Product No. 4583, Miles, Inc., Diagnostics Division, Elkhart, IN 46515).
4. Gently mix the cells with the O.C.T. Compound. Snap freeze if cryomolds
are not available. If cryomolds are available, continue as in item 5.
5. Pour O.C.T.-suspended cells into plastic 10 X 10 X 5 mm Cryomold. Snap
freeze in liquid nitrogen or on dry ice.
6. Place the Cryomolds containing the suspended cells into Ziploc plastic
bags. Ship on dry ice.
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