Re: [Histonet] Von Kossa staining
Charles, I am going to date myself and tell you that we placed the glass coplin jar on the back of the embedder (or in the water bath) added a bankers light and wrapped it with foil for 1 hr. Sincerely, Toysha N. Mayer, DHSc, MBA, HT (ASCP) Asst Professor/ Assoc Program Director HTL Program UTMDACC tnma...@mdanderson.edu off cell: 832-710-1837 off: 713-563-3481 Today's Topics: 1. Von Kossa staining (Charles Riley) 2. Re: Von Kossa staining (Debra Siena) 3. Re: Von Kossa staining (John Kiernan) 4. Re: Von Kossa staining (Bernice Frederick) 5. Re: Von Kossa staining (Mac Donald, Jennifer) -- Message: 1 Date: Wed, 26 Jul 2023 14:23:16 -0400 From: Charles Riley To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Von Kossa staining Message-ID: Content-Type: text/plain; charset="UTF-8" Can anyone out there who performs Von Kossa staining provide me with any guidelines or suggestions for the light source to use for the Silver nitrate activation? Is a standard handheld black light strong enough or does it need to be a UV sanitizing strength light if using UV versus incandescent bulb exposure? -- The information contained in this e-mail message may be privileged, confidential, and/or protected from disclosure. This e-mail message may contain protected health information (PHI); dissemination of PHI should comply with applicable federal and state laws. If you are not the intended recipient, or an authorized representative of the intended recipient, any further review, disclosure, use, dissemination, distribution, or copying of this message or any attachment (or the information contained therein) is strictly prohibited. If you think that you have received this e-mail message in error, please notify the sender by return e-mail and delete all references to it and its contents from your systems. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Von Kossa staining
I have used a UV light like the ones for indoor plants and one that is in the fume hood for disinfection. Both proved successful. Jennifer -Original Message- From: Charles Riley via Histonet Sent: Wednesday, July 26, 2023 11:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Von Kossa staining EXTERNAL SENDER - Exercise caution with requests, links, and attachments. Can anyone out there who performs Von Kossa staining provide me with any guidelines or suggestions for the light source to use for the Silver nitrate activation? Is a standard handheld black light strong enough or does it need to be a UV sanitizing strength light if using UV versus incandescent bulb exposure? ___ 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] Von Kossa staining
I use a 60 watt bulb and it works fine. Bernice -Original Message- From: John Kiernan via Histonet Sent: Wednesday, July 26, 2023 11:51 PM To: Charles Riley ; histonet@lists.utsouthwestern.edu; dsiena Subject: Re: [Histonet] Von Kossa staining Charles, A handheld light of any kind isn't really suitable because you would have to hold it over the slides for 15 to 60 minutes, according to which variant of the von Kossa method you plan to use (see Lillie & Fullmer 1976 Histopathologic Technic ... 4th ed. pp 539-541). An anglepoise lamp with an old-fashioned 100W bulb is OK, and so is a sunny windowsill. Silver salts absorb at the blue end of the spectrum, so a fluorescent light should be more efficient than an incandescent bulb. If no bright light source is available, it's possible to chemically reduce the silver phosphate and/or carbonate to black colloidal silver, with a traditional photographic developer. The method of Rungby et al.1993 may be better than other post-reduction methods (https://urldefense.com/v3/__https://scholar.google.ca/scholar?hl=en_sdt=0*2C5=rungby*1993*calcium*deposits=rungby*1993__;JSsrKys!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfChgoE5Zi$ ). I never tried it, but Rungby's paper has collected 104 citations, which is very good for a paper in our field. The von Kossa technique is simply explained in my Histological and Histochemical Methods textbook, 5th edn (2015). The book costs less than 1ml of any antibody. Enough said! John John A. Kiernan Emeritus, Anatomy & Cell Biology, University of Western Ontario London, Canada https://urldefense.com/v3/__https://www.schulich.uwo.ca/anatomy/people/bios/emeriti/kiernan_john.html__;!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfCiLlysL2$ Also Secretary, Biological Stain Commission, Inc. https://urldefense.com/v3/__https://biologicalstaincommission.org__;!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfCtBsy2U8$ = = = -Original Message- From: Charles Riley via Histonet Sent: Wednesday, July 26, 2023 1:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Von Kossa staining *** Externally sourced email message *** Can anyone out there who performs Von Kossa staining provide me with any guidelines or suggestions for the light source to use for the Silver nitrate activation? Is a standard handheld black light strong enough or does it need to be a UV sanitizing strength light if using UV versus incandescent bulb exposure? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu https://urldefense.com/v3/__http://lists.utsouthwestern.edu/mailman/listinfo/histonet__;!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfCgGN6_75$ CAUTION: This email was originated from outside StatLab and contains links. Do not click links or open attachments unless you recognize the sender and you know the content is safe. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu https://urldefense.com/v3/__http://lists.utsouthwestern.edu/mailman/listinfo/histonet__;!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfCgGN6_75$ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu https://urldefense.com/v3/__http://lists.utsouthwestern.edu/mailman/listinfo/histonet__;!!Dq0X2DkFhyF93HkjWTBQKhk!SGP1gx0IglGW5nnid6Tbv6aRuFAWHsIiz6kn3jDzSw7njXRiBLURbNbbv4YCJKl-Jbcpm60EOukGFQIdqh2rLhqFIgrWGAWfCgGN6_75$ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Von Kossa staining
Charles, A handheld light of any kind isn't really suitable because you would have to hold it over the slides for 15 to 60 minutes, according to which variant of the von Kossa method you plan to use (see Lillie & Fullmer 1976 Histopathologic Technic ... 4th ed. pp 539-541). An anglepoise lamp with an old-fashioned 100W bulb is OK, and so is a sunny windowsill. Silver salts absorb at the blue end of the spectrum, so a fluorescent light should be more efficient than an incandescent bulb. If no bright light source is available, it's possible to chemically reduce the silver phosphate and/or carbonate to black colloidal silver, with a traditional photographic developer. The method of Rungby et al.1993 may be better than other post-reduction methods (https://scholar.google.ca/scholar?hl=en_sdt=0%2C5=rungby+1993+calcium+deposits=rungby+1993). I never tried it, but Rungby's paper has collected 104 citations, which is very good for a paper in our field. The von Kossa technique is simply explained in my Histological and Histochemical Methods textbook, 5th edn (2015). The book costs less than 1ml of any antibody. Enough said! John John A. Kiernan Emeritus, Anatomy & Cell Biology, University of Western Ontario London, Canada https://www.schulich.uwo.ca/anatomy/people/bios/emeriti/kiernan_john.html Also Secretary, Biological Stain Commission, Inc. https://biologicalstaincommission.org = = = -Original Message- From: Charles Riley via Histonet Sent: Wednesday, July 26, 2023 1:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Von Kossa staining *** Externally sourced email message *** Can anyone out there who performs Von Kossa staining provide me with any guidelines or suggestions for the light source to use for the Silver nitrate activation? Is a standard handheld black light strong enough or does it need to be a UV sanitizing strength light if using UV versus incandescent bulb exposure? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CAUTION: This email was originated from outside StatLab and contains links. Do not click links or open attachments unless you recognize the sender and you know the content is safe. ___ 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] Von Kossa staining
Hi Charles, I can't answer your question about UV light as I've never tried it except with incandescent lights. However, I do know that you could use natural sunlight as well to develop the silver color change. I hope that helps, Thanks Debbie Siena, HT(ASCP)QIHC Director of Scientific Affairs Mobile:817-994-9407 Dallas, TX | Baltimore, MD | Mt. Vernon, WA -Original Message- From: Charles Riley via Histonet Sent: Wednesday, July 26, 2023 1:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Von Kossa staining *** Externally sourced email message *** Can anyone out there who performs Von Kossa staining provide me with any guidelines or suggestions for the light source to use for the Silver nitrate activation? Is a standard handheld black light strong enough or does it need to be a UV sanitizing strength light if using UV versus incandescent bulb exposure? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CAUTION: This email was originated from outside StatLab and contains links. Do not click links or open attachments unless you recognize the sender and you know the content is safe. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Von Kossa staining
Can anyone out there who performs Von Kossa staining provide me with any guidelines or suggestions for the light source to use for the Silver nitrate activation? Is a standard handheld black light strong enough or does it need to be a UV sanitizing strength light if using UV versus incandescent bulb exposure? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Von Kossa staining on PMMA sections
The standard Von Kossa silver stain for calcium calls for 20 minutes in the silver nitrate solution under UV light. There is a modified Von Kossa for plastic embedded bone sections, which is identical except it calls for a minimum of 6 hours in the silver nitrate solution under UV. Does anyone know why such a long staining time is recommended? Visually the calcium in the bone sections turns black within 20 minutes, so why is so much additional time needed? Thanks. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Von Kossa staining on PMMA sections
Paul, For MMA embedded specimens (MMA +DBP), I first deplastify my sections, hydrate to water, stain in 5% silver nitrate solution for 5 minutes (in the dark), wash times three changes in DI H2O (in the dark), develop in sodium-carbonate formaldehyde solution for 2 minutes (in the dark), wash times two in DI H2O (back under normal lighting conditions), then stop the reaction in sodium thiosulfate + potassium ferricyanide solution for 30 seconds, and immediately rinse in running tap water for 15 minutes. The Von Kossa reaction results from process above then yields black mineralized bone. After the tap water rinse, I generally counterstain with 2% MacNeal's tetrachrome for 5 minutes, rinse in DI H2O, and dehydrate to xylenes to coverslip. This then reveals immature bone formation or osteoid = grayish or jaded green, growth plate cartilage = purple, osteoblasts = blue, osteoclasts = blue-green, bloods cells = greenish, etc. Feel free to contact me if you have any additional questions. Jack Date: Thu, 12 Mar 2009 11:19:49 -0400 From: pmonf...@lifespan.org To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Von Kossa staining on PMMA sections The standard Von Kossa silver stain for calcium calls for 20 minutes in the silver nitrate solution under UV light. There is a modified Von Kossa for plastic embedded bone sections, which is identical except it calls for a minimum of 6 hours in the silver nitrate solution under UV. Does anyone know why such a long staining time is recommended? Visually the calcium in the bone sections turns black within 20 minutes, so why is so much additional time needed? Thanks. ___ 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
