We cryoprotect our livers in 20% sucrose overnight at 4C after fixation in 4% paraformaldehyde, then freeze in thawing isopentane (freeze it over liquid nitrogen, lift clear and use it as it melts, but before it has melted completely). Store if necessary at -80C. Section onto charged (Plus) slides and stain in the normal way. I cannot see why yours are coming off, but suggest you try plus slides, then they won't.
I can't think that the cryoprotection makes any difference to whether they fall off or not, but I think the sections are easier to cut after cryoprotection than without it. I have been given livers treated both ways. Plus slides: Superfrost plus, available from all good histology suppliers; ours come from VWR. Good luck. Miss Margaret Blount Histology Manager Metabolic Research Laboratories Level 4 Institute of Metabolic Science Box 289, Addenbrooke's Hospital Hills Road, Cambridge, CB2 0QQ Tel 01223 769061/336079 -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of kyoung LEE Sent: 14 April 2011 15:07 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] [Help] oil red O stain in fattty change liver Dear sirs. I don't know whether I send this question to you. For several weeks, I'm setting oil red O stain in fattty change liver (mouse). Could you review my protocol? 1. Murine tissue,livers, is fixed with 10% NBF(10% neutral-buffered formalin). 2. Make frozen block 3. Blocks were cut 5um in crystat and dried for 1~2H at RT. 4. store at -20 5. let it dry for 30mins at RT 6. immerse it in cold 10% NBF for 10mins 7. staining After staining, my section were fallen off and seperated. I search many protocols. Some suggest additional fixation is necessary as soon as cutting. Others suggest prefixed tissue is not necessary additional fixation. Plz send me your comments. _______________________________________________ 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