[Histonet] Remove OCT from frozen tissue block
Dear histoneters, We got mice brains that were initially sucrose-cryoprotected and then embedded in OCT - flash frozen and stored at -80°C. These tissues were initially prepared for subsequent cryostat sectioning but due to technical considerations we need to cut them using a sliding microtome to get thick (40µm) floating sections. We therefore would like to remove OCT before cutting. As OCT is water soluble it is expected that placing the OCT blocks in buffer would help removing the embedding media but thawing the tissue and then re-freezing it on the microtome stage might not be the best way to proceed (expected formation of ice crystals with known associated histological artefacts)... We would appreciate any alternative solutions. Thanks! Benoît ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Triton x-100 stability
Dear Histoneters, We have recently experienced some troubles in carrying out routine immuno against nuclear fos protein (unexpected weak signal). We have troubleshooted all what was possible to troubleshoot (use of 3 different primary antibodies batches, two secondary ab, new buffers, change the revelation kit etc.). Immunos directed against other epitopes (eg amyloid-beta) give satisfactory results but the IHC in trouble, that targets intranuclear epitopes, is still defective. I suspect now that the non-ionic surfactant we are using (Triton x-100) could have been degraded by high temperatures we had for several weeks in the lab (due to a central heating problem in our building). I made some temp. measurements in the reactives cupboard (around 35-40°C). Does anyone have any information on Triton x-100 stability? According to manufacturers' recommendations, the product should be stored at RT but I did not succeed to get more precise information. I thank you in advance for all comments, suggestions and alternatives. B. Delatour -- Laboratoire de Neurobiologie de l'Apprentissage, de la Mémoire de la Communication, NAMC, CNRS UMR 8620, Bât 446 Université Paris-Sud 91405 Orsay Cedex, FRANCE Email benoit.delat...@u-psud.fr Webhttp://www.namc.u-psud.fr If you think research is expensive, try disease. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: staining for manganese
Hi, There is a Timm stain - like protocol from Angenstein (Manganese-enhanced MRI reveals structural and functional changes in the cortex of Bassoon mutant mice. Angenstein F, Niessen HG, Goldschmidt J, Lison H, Altrock WD, Gundelfinger ED, Scheich H. Cereb Cortex. 2007 Jan;17(1):28-36) using Na2S that shows some interesting results with manganese autometallography. The problem, as mentionned by RJ Buesa, concerns the specificity of the staining. Using this method in the past I observed staining increase in the brain following MnCl injection. Regards, Benoît Delatour -- Message: 1 Date: Mon, 26 Jan 2009 18:58:34 +0900 From: Gerard Spoelstra g.spoels...@murdoch.edu.au Subject: [Histonet] staining for manganese To: histonet@lists.utsouthwestern.edu Message-ID: cf0145ebf1eb4c4e82768d82886a0c9b8fb...@pluto.ad.murdoch.edu.au Content-Type: text/plain; charset=iso-8859-1 Hi everybody, I have some fish specimens which have been exposed to toxic levels of manganese. The researcher is hoping to see manganese localised on the gill. Lillie mentions the use of benzidine to stain for manganese. I will try DAB, but my searching on the net has drawn a blank. Gerard Spoelstra veterinary histology Murdoch University Western Australia ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Epson v750 pro scanner
Dear Histoneters, Has anyone ever used the Epson v750 pro flatbed scanner to digitize microscopic slides? In the past we get used to a Nikon coolscan scanner and got nice results. We then bought the Epson scanner because of its high spatial resolution (6400 optical dpi) but did not obtain satisfactory images, principally because of bad focusing troubles. Does anyone has a good experience / expertise with this scanner and an efficient protocol to numerize slides with this device? Thanks to all, B. Delatour ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
