Re: [Histonet] Butyl alcohol
As an additional note: paraffin oil = mineral oil. René J. --- On Tue, 3/9/10, Michael Folsom mwfol...@rgbio.com wrote: From: Michael Folsom mwfol...@rgbio.com Subject: Re: [Histonet] Butyl alcohol To: Krueger, Todd todd.krue...@bsci.com Cc: histonet@lists.utsouthwestern.edu Date: Tuesday, March 9, 2010, 11:17 AM Hi: I'm not sure if this helps but Botanists have long used tert-butanol (not N-butanol) to embed tissue in paraffin. This classic protocol was popularized by D. A. Johansen in the mid 1930's and involves transition from water to alcohol to t-butanol (I can supply more detailed info if you need it). Once in 100% t-butanol tissue is transitioned to light paraffin oil. The final stage involves filling some large shell vials 1/3 full with paraffin and letting them harden. Tissue plus paraffin oil is added to the vials (volume of paraffin oil + tissue should be be approximately equal to the amount of paraffin). Vials are now placed in an oven for paraffin to melt and infiltration to begin. Besides doing enough changes of paraffin to infiltrate the tissue and remove all the paraffin oil that's about it. From there its the old embed, section and enjoy! Hope this helps - Mike Folsom Rio Grande Biological mwfol...@rgbio.com On Mon, 2010-03-08 at 09:00 -0600, Krueger, Todd wrote: Does anyone have a procedure for processing with N-butyl Alcohol. We need to find a procedure w/o xylene. Thanks Todd Krueger HTL(ASCP)CM Boston Scientific 2 Scimed Place, P121 Osseo, MN 55311 Phone: 763-694-5709 Fax: 763-694-5505 e-mail: todd.krue...@bsci.com ___ 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Butyl alcohol
Use 2-propanol instead. It is better and much cheaper. René J.U --- On Mon, 3/8/10, Krueger, Todd todd.krue...@bsci.com wrote: From: Krueger, Todd todd.krue...@bsci.com Subject: [Histonet] Butyl alcohol To: histonet@lists.utsouthwestern.edu Date: Monday, March 8, 2010, 10:00 AM Does anyone have a procedure for processing with N-butyl Alcohol. We need to find a procedure w/o xylene. Thanks Todd Krueger HTL(ASCP)CM Boston Scientific 2 Scimed Place, P121 Osseo, MN 55311 Phone: 763-694-5709 Fax: 763-694-5505 e-mail: todd.krue...@bsci.com ___ 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
Re: [Histonet] Butyl alcohol
Dear Todd Krueger, There are three isomeric butyl alcohols (primary, secondary and tertiary) with different physical properties. and different uses in histology for primary and tertiary. I have used tertiary butyl alcohol (= t-butanol = 2-methylpropan-2-ol) a few times for processing into paraffin. It mixes with with both water and wax. It boils at 83C so it's slightly less of a fire hazard than ethanol. It doesn't form explosive peroxides with long storage, which makes it safer than dioxane and tetrahydrofuran, two other universal solvents that have been used for combined dehydration and clearing. t-Butanol doesn't have an offensive odour, but it is solid below 25C, which is inconvenient, and it's quite a bit more expensive than more commonly used solvents such as ethanol, isopropanol and xylene. t-Butanol was introduced as a combined dehydration and clearing agent by Larbaud (1921) Compt. Rend. Acad. Sci. 172:1317-1319. It is used more in plant than in animal histology. Primary butyl alcohol (n-butanol) is liquid at ordinary temperatures. It is only partly miscible with water, but miscible with ethanol-water mixtures and with paraffin. It has been recommended for transitioning to wax in procedures claimed to reduce hardening of wood (Zirkle 1930, Science 71:103-104) and insect specimens (Stiles 1934, Stain Technol. 9:97-100). Freeze-substitution into n-butanol can be followed by paraffin embedding because this alcohol is a liquid from -90 to +117C. I haven't tried any of these methods. Another use of n-butanol is in dehydration of sections stained with dyes that are easily extracted by water or water-ethanol mixtures. I have lots of experience in this area. A major disadvantage of n-butanol, for any application, is its vapour. It doesn't smell nasty but it makes you cough. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: Krueger, Todd todd.krue...@bsci.com Date: Wednesday, March 3, 2010 17:08 Subject: [Histonet] Butyl alcohol To: histonet@lists.utsouthwestern.edu Has anyone tried using butyl alcohol as a dehydrating and clearing agent on there tissues? What are some pros and cons people have experienced with using Butyl alcohol? Todd Krueger HTL(ASCP)CM Boston Scientific 2 Scimed Place, P121 Osseo, MN 55311 Phone: 763-694-5709 Fax: 763-694-5505 e-mail: todd.krue...@bsci.com ___ 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
