Hi Tim, In our lab, we use positive charged slides for all special stains. We clean our glassware using a "low-sudsing" soap with bleach (~1 cup) added and then rinsed 3 times using distilled water. When we make up the formaldehyde solution, we consider the 40% to be 100% in making the dilution.
hope this helped, Lynette Lynette Pavelich, HT(ASCP) Histology Supervisor Hurley Medical Center One Hurley Plaza Flint, MI 48503 ph: 810.262.9948 mobile: 810.444.7966 ________________________________________ From: [email protected] [[email protected]] on behalf of Tim Wheelock [[email protected]] Sent: Tuesday, July 24, 2012 3:36 PM To: [email protected] Subject: [Histonet] Bielschowsky questions, Luxol Fast Blue answers, and Protocol Drift. Hi Everyone: I had a few questions regarding Bielschowsky silver stains. (1) What adhesive (if any) or type of slide do you use for the stain? (2) How do you clean the glassware? (3) When diluting the 40% formaldehyde when making up the developer, do you consider the 40% formaldehyde as 100% and then dilute it down by using 1 part formaldehyde and 9 parts distilled water? Or do you assume the 40% formaldehyde is 40% and then dilute it down using 1 part formaldehyde and 3 parts distilled water? (My protocol may have inadvertently changed from the first method to the second; I am not sure.) By the way, I want to thank everyone for helping me solve the problem of my Luxol Fast Blue staining the myelin too lightly. I discovered that somehow, I had started adding twice the amount of acetic acid to the Luxol staining solution as I should of. (This protocol "drift", where a mistake can actually find its way into a written protocol, can be a real problem in a lab, especially when working for years by oneself, as I have) . But I also found that even reducing the acetic acid, while helping a lot, did not completely fix the problem. I needed to switch from staining the tissue for 2 hours at 60C to a full over-night (why I never needed to switch times before is a mystery). That did the trick beautifully. The myelin is staining perfectly again. Thanks again, Tim Wheelock Harvard Brain Bank Belmont, MA _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
