Re: [Histonet] Prussian Blue Reaction
Thank you Tony. -Original Message- From: Tony Henwood (SCHN) Sent: Monday, June 7, 2021 4:25 PM To: Mac Donald, Jennifer ; Jay Lundgren Cc: John Kiernan ; 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. The following might be useful: Iron Histochemistry - A Review It is convenient to divide iron-containing complexes in human tissues into two categories: those in which the iron is loosely bound to proteins and easily released by mild acid treatment (eg hemosiderin) and those in which the iron is more strongly bound (masked iron) and cannot be released by mild acid hydrolysis (eg haemoglobin) (1). Iron in the body is stored in the forms of hemosiderin (ferric hydroxide polymer) or ferritin (a ferrous iron-protein complex) (1). Iron in tissues occurs mainly in the ferric state (2,3). The reactions that have been used for the detection of iron in tissues include (2-5): 1. The Quincke reaction using ammonium sulphide 2. The Perls reaction, using ferrocyanide, for ferric and the Turnbull Blue reaction, using ferricyanide for ferrous iron. 3. Coloured lakes, eg haematoxylin (Mallory's Method) 4. Coloured precipitates with organic chemicals not classified as dyes (eg bathophenanthroline). Ferric iron may be converted into ferrous iron by ammonium sulphide (Quinke's reaction) and the ferrous sulphide thus formed can then be demonstrated using the Turnbull blue reaction (3,5). Some iron-containing compounds (hemoglobin, malaria pigment, formalin pigment) do not react with the Perl's method because the iron is present in bound form. These compounds can be unmasked using hydrogen peroxide and then demonstrated using the Perl's reaction (1). Interestingly, it is possible to remove excess iron pigment from tissue sections. Iron can be removed by (5): . 15 min in 1% sodium dithionite in 0.1M acetate-HCl buffer (pH 4.5) . 3 hours in 2.4N HCl . 30 min in 3.7N H2SO4 . 15 min in 5% Oxalic acid Heavily pigmented tissues may need to have these times extended (5). References 1. Barka, T., Anderson, P.J., (1963) "Histochemistry: Theory, practice and bibliography" Harper & Row Publishers Inc, New York, p172-174. 2. Davenport, H.A., (1961) "Histological and Histochemical Technics" W.B. Saunders Co., Philadelphia, 280-284. 3. Gabe, M., (1976) "Histological Techniques" Masson, Paris, p311-317. 4. Lynch, M.J., Raphael, S., etal "Medical Laboratory Technology and Clinical Pathology" 2nd Ed, W.B Saunder Co, Philadelphia, p1135-1136. 5. Morton, D., (1978) "A comparison of iron histochemical methods for use on glycol methacrylate embedded tissues" Stain Tech 53(4):217-223. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: Mac Donald, Jennifer via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Tuesday, 8 June 2021 6:41 AM To: Jay Lundgren Cc: histonet@lists.utsouthwestern.edu; John Kiernan Subject: Re: [Histonet] Prussian Blue Reaction Thank you. From: Jay Lundgren Sent: Monday, June 7, 2021 1:35 PM To: Mac Donald, Jennifer Cc: John Kiernan ; Gudrun Lang ; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Supposed to be insoluble. Try potassium permanganate followed by oxalaic acid. But book says insoluble. On Monday, June 7, 2021, Mac Donald, Jennifer via Histonet mailto:histonet@lists.utsouthwestern.edu>> wrote: The instrument malfunction and it was overstained. From: John Kiernan mailto:jkier...@uwo.ca>> Sent: Monday, June 7, 2021 9:50 AM To: Gudrun Lang mailto:gu.l...@gmx.at>>; Mac Donald, Jennifer mailto:jmacdon...@mtsac.edu>> Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Overstained? Doesn't that mean the tissue contains a lot of iron and you are seeing where it is - which was the reason for doing Prussian blue histochemistry. Gudrun Lang correctly says that mineral acids won't remove it. Oxalic acid is said to dissolve Prussian blue (? by chelation); I've never tried this. If it works, you will no longer see where the iron is. To see features other than the distribution of iron, why not just stain another section from
Re: [Histonet] Prussian Blue Reaction
The following might be useful: Iron Histochemistry - A Review It is convenient to divide iron-containing complexes in human tissues into two categories: those in which the iron is loosely bound to proteins and easily released by mild acid treatment (eg hemosiderin) and those in which the iron is more strongly bound (masked iron) and cannot be released by mild acid hydrolysis (eg haemoglobin) (1). Iron in the body is stored in the forms of hemosiderin (ferric hydroxide polymer) or ferritin (a ferrous iron-protein complex) (1). Iron in tissues occurs mainly in the ferric state (2,3). The reactions that have been used for the detection of iron in tissues include (2-5): 1. The Quincke reaction using ammonium sulphide 2. The Perls reaction, using ferrocyanide, for ferric and the Turnbull Blue reaction, using ferricyanide for ferrous iron. 3. Coloured lakes, eg haematoxylin (Mallory's Method) 4. Coloured precipitates with organic chemicals not classified as dyes (eg bathophenanthroline). Ferric iron may be converted into ferrous iron by ammonium sulphide (Quinke's reaction) and the ferrous sulphide thus formed can then be demonstrated using the Turnbull blue reaction (3,5). Some iron-containing compounds (hemoglobin, malaria pigment, formalin pigment) do not react with the Perl's method because the iron is present in bound form. These compounds can be unmasked using hydrogen peroxide and then demonstrated using the Perl's reaction (1). Interestingly, it is possible to remove excess iron pigment from tissue sections. Iron can be removed by (5): . 15 min in 1% sodium dithionite in 0.1M acetate-HCl buffer (pH 4.5) . 3 hours in 2.4N HCl . 30 min in 3.7N H2SO4 . 15 min in 5% Oxalic acid Heavily pigmented tissues may need to have these times extended (5). References 1. Barka, T., Anderson, P.J., (1963) "Histochemistry: Theory, practice and bibliography" Harper & Row Publishers Inc, New York, p172-174. 2. Davenport, H.A., (1961) "Histological and Histochemical Technics" W.B. Saunders Co., Philadelphia, 280-284. 3. Gabe, M., (1976) "Histological Techniques" Masson, Paris, p311-317. 4. Lynch, M.J., Raphael, S., etal "Medical Laboratory Technology and Clinical Pathology" 2nd Ed, W.B Saunder Co, Philadelphia, p1135-1136. 5. Morton, D., (1978) "A comparison of iron histochemical methods for use on glycol methacrylate embedded tissues" Stain Tech 53(4):217-223. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message- From: Mac Donald, Jennifer via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Tuesday, 8 June 2021 6:41 AM To: Jay Lundgren Cc: histonet@lists.utsouthwestern.edu; John Kiernan Subject: Re: [Histonet] Prussian Blue Reaction Thank you. From: Jay Lundgren Sent: Monday, June 7, 2021 1:35 PM To: Mac Donald, Jennifer Cc: John Kiernan ; Gudrun Lang ; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Supposed to be insoluble. Try potassium permanganate followed by oxalaic acid. But book says insoluble. On Monday, June 7, 2021, Mac Donald, Jennifer via Histonet mailto:histonet@lists.utsouthwestern.edu>> wrote: The instrument malfunction and it was overstained. From: John Kiernan mailto:jkier...@uwo.ca>> Sent: Monday, June 7, 2021 9:50 AM To: Gudrun Lang mailto:gu.l...@gmx.at>>; Mac Donald, Jennifer mailto:jmacdon...@mtsac.edu>> Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Overstained? Doesn't that mean the tissue contains a lot of iron and you are seeing where it is - which was the reason for doing Prussian blue histochemistry. Gudrun Lang correctly says that mineral acids won't remove it. Oxalic acid is said to dissolve Prussian blue (? by chelation); I've never tried this. If it works, you will no longer see where the iron is. To see features other than the distribution of iron, why not just stain another section from the block with a general-purpose stain like Giemsa or H? John Kiernan London, Canada = = = From: Mac Donald, Jennifer via Histonet mailto:histonet@lists.utsouthwestern.edu><mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>>> Sent: June 7, 2021 12:47 AM To: Gudrun Lang mailto
Re: [Histonet] Prussian Blue Reaction
Thank you. From: Jay Lundgren Sent: Monday, June 7, 2021 1:35 PM To: Mac Donald, Jennifer Cc: John Kiernan ; Gudrun Lang ; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Supposed to be insoluble. Try potassium permanganate followed by oxalaic acid. But book says insoluble. On Monday, June 7, 2021, Mac Donald, Jennifer via Histonet mailto:histonet@lists.utsouthwestern.edu>> wrote: The instrument malfunction and it was overstained. From: John Kiernan mailto:jkier...@uwo.ca>> Sent: Monday, June 7, 2021 9:50 AM To: Gudrun Lang mailto:gu.l...@gmx.at>>; Mac Donald, Jennifer mailto:jmacdon...@mtsac.edu>> Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Overstained? Doesn't that mean the tissue contains a lot of iron and you are seeing where it is - which was the reason for doing Prussian blue histochemistry. Gudrun Lang correctly says that mineral acids won't remove it. Oxalic acid is said to dissolve Prussian blue (? by chelation); I've never tried this. If it works, you will no longer see where the iron is. To see features other than the distribution of iron, why not just stain another section from the block with a general-purpose stain like Giemsa or H? John Kiernan London, Canada = = = From: Mac Donald, Jennifer via Histonet mailto:histonet@lists.utsouthwestern.edu><mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>>> Sent: June 7, 2021 12:47 AM To: Gudrun Lang mailto:gu.l...@gmx.at><mailto:gu.l...@gmx.at<mailto:gu.l...@gmx.at>>> Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu><mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>> mailto:histonet@lists.utsouthwestern.edu><mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>>> Subject: Re: [Histonet] Prussian Blue Reaction The tissue was overstained and the blue was interfering with interpretation -Original Message- From: Gudrun Lang mailto:gu.l...@gmx.at><mailto:gu.l...@gmx.at<mailto:gu.l...@gmx.at>>> Sent: Sunday, June 6, 2021 2:18 AM To: Mac Donald, Jennifer mailto:jmacdon...@mtsac.edu><mailto:jmacdon...@mtsac.edu<mailto:jmacdon...@mtsac.edu>>> Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu><mailto:histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>> Subject: AW: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Hi Jennifer, Why do you want to reduce the staining? I ask, because the impact of hydrochloric acid on the tissue may influence the following results anyway. I think, that the prussian blue pigment cannot be removed in an easy way. It is resistent to solvents and mineral acids. https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian-Bluedata=04%7C01%7Cjmacdonald%40mtsac.edu%7C0fbc82a2b13749a4222608d928cbfe52%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637585679205067185%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000sdata=KjvijcfrVPGZKGsAn6qX5rMKtulHpmsAzqHEkwz%2B96Y%3Dreserved=0<https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian-Blue=04%7C01%7Cjmacdonald%40mtsac.edu%7C684bae0e8884489b44cb08d929f3cae3%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637586952354386054%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000=d22NsJ2m3eoU6bFZ3mvyb6rIaFr%2BcMu4tuS40b1hcsk%3D=0><https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian-Blue=04%7C01%7Cjmacdonald%40mtsac.edu%7C2eeb915058594b1d14b508d929d455c4%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637586814671423758%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000=xKy3v4qvO4spk0zrRxXWZfcsHWxbQC%2FIQ1FbpMB390Y%3D=0<https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian-Blue=04%7C01%7Cjmacdonald%40mtsac.edu%7C684bae0e8884489b44cb08d929f3cae3%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637586952354386054%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000=d22NsJ2m3eoU6bFZ3mvyb6rIaFr%2BcMu4tuS40b1hcsk%3D=0>> -for-Solvent-Based-Inks.html On the other hand, if th
Re: [Histonet] Prussian Blue Reaction
Supposed to be insoluble. Try potassium permanganate followed by oxalaic acid. But book says insoluble. On Monday, June 7, 2021, Mac Donald, Jennifer via Histonet < histonet@lists.utsouthwestern.edu> wrote: > The instrument malfunction and it was overstained. > > From: John Kiernan > Sent: Monday, June 7, 2021 9:50 AM > To: Gudrun Lang ; Mac Donald, Jennifer < > jmacdon...@mtsac.edu> > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Prussian Blue Reaction > > EXTERNAL SENDER- Exercise caution with requests, links, and attachments. > Overstained? Doesn't that mean the tissue contains a lot of iron and you > are seeing where it is - which was the reason for doing Prussian blue > histochemistry. Gudrun Lang correctly says that mineral acids won't remove > it. Oxalic acid is said to dissolve Prussian blue (? by chelation); I've > never tried this. If it works, you will no longer see where the iron is. To > see features other than the distribution of iron, why not just stain > another section from the block with a general-purpose stain like Giemsa or > H? > John Kiernan > London, Canada > = = = > > From: Mac Donald, Jennifer via Histonet <mailto:histonet@lists.utsouthwestern.edu>> > Sent: June 7, 2021 12:47 AM > To: Gudrun Lang mailto:gu.l...@gmx.at>> > Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists. > utsouthwestern.edu> mailto:hist > o...@lists.utsouthwestern.edu>> > Subject: Re: [Histonet] Prussian Blue Reaction > > The tissue was overstained and the blue was interfering with interpretation > > -Original Message- > From: Gudrun Lang mailto:gu.l...@gmx.at>> > Sent: Sunday, June 6, 2021 2:18 AM > To: Mac Donald, Jennifer mailto:jmacdon...@mtsac.edu > >> > Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists. > utsouthwestern.edu> > Subject: AW: [Histonet] Prussian Blue Reaction > > EXTERNAL SENDER- Exercise caution with requests, links, and attachments. > > Hi Jennifer, > Why do you want to reduce the staining? > > I ask, because the impact of hydrochloric acid on the tissue may influence > the following results anyway. > I think, that the prussian blue pigment cannot be removed in an easy way. > It is resistent to solvents and mineral acids. > https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww. > epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian- > Bluedata=04%7C01%7Cjmacdonald%40mtsac.edu% > 7C0fbc82a2b13749a4222608d928cbfe52%7Ccc4d4bf20a9e4240aedea7d1d688 > f935%7C0%7C0%7C637585679205067185%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAw > MDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000sdata= > KjvijcfrVPGZKGsAn6qX5rMKtulHpmsAzqHEkwz%2B96Y%3Dreserved=0< > https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww. > epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian- > Blue=04%7C01%7Cjmacdonald%40mtsac.edu%7C2eeb915058594b1d14b508d929d4 > 55c4%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637586814671423758% > 7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik > 1haWwiLCJXVCI6Mn0%3D%7C1000=xKy3v4qvO4spk0zrRxXWZfcsHWxbQC > %2FIQ1FbpMB390Y%3D=0> > -for-Solvent-Based-Inks.html > > On the other hand, if the blue colour doesn't interfere with your > following staining, you can try to simple make a "double stain". > > Regards > Gudrun > > -Ursprüngliche Nachricht- > Von: Mac Donald, Jennifer via Histonet > [mailto:histonet@lists.utsouthwestern.edu] > Gesendet: Sonntag, 6. Juni 2021 06:34 > An: histonet@lists.utsouthwestern.edu<mailto:histonet@lists. > utsouthwestern.edu> > Betreff: [Histonet] Prussian Blue Reaction > > > Does anyone know of a way to remove/reduce the Prussian blue reaction? > Thanks, > Jennifer > > > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu<mailto:Histonet@lists.utsouthwestern.edu > > > https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists. > utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonet > data=04%7C01%7Cjmacdonald%40mtsac.edu%7C0fbc82a2b13749a4222608d928cbfe52% > 7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637585679205067185%7CUnknown% > 7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik > 1haWwiLCJXVCI6Mn0%3D%7C2000sdata=gRrUmRDEU3BfcA0rEgQOgBvPHIQ05I > RM6WozVZiiR1g%3Dreserved=0<https://nam12.safelinks. > protection.outlook.com/?url=http%3A%2F%2Flists. > utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonet=04% > 7C01%7Cjmacdonald%40mtsac.edu%7C2eeb915058594b1d14b508d929d455c4% > 7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C6375
Re: [Histonet] Prussian Blue Reaction
The instrument malfunction and it was overstained. From: John Kiernan Sent: Monday, June 7, 2021 9:50 AM To: Gudrun Lang ; Mac Donald, Jennifer Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Overstained? Doesn't that mean the tissue contains a lot of iron and you are seeing where it is - which was the reason for doing Prussian blue histochemistry. Gudrun Lang correctly says that mineral acids won't remove it. Oxalic acid is said to dissolve Prussian blue (? by chelation); I've never tried this. If it works, you will no longer see where the iron is. To see features other than the distribution of iron, why not just stain another section from the block with a general-purpose stain like Giemsa or H? John Kiernan London, Canada = = = From: Mac Donald, Jennifer via Histonet mailto:histonet@lists.utsouthwestern.edu>> Sent: June 7, 2021 12:47 AM To: Gudrun Lang mailto:gu.l...@gmx.at>> Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> mailto:histonet@lists.utsouthwestern.edu>> Subject: Re: [Histonet] Prussian Blue Reaction The tissue was overstained and the blue was interfering with interpretation -Original Message- From: Gudrun Lang mailto:gu.l...@gmx.at>> Sent: Sunday, June 6, 2021 2:18 AM To: Mac Donald, Jennifer mailto:jmacdon...@mtsac.edu>> Cc: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Subject: AW: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Hi Jennifer, Why do you want to reduce the staining? I ask, because the impact of hydrochloric acid on the tissue may influence the following results anyway. I think, that the prussian blue pigment cannot be removed in an easy way. It is resistent to solvents and mineral acids. https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian-Bluedata=04%7C01%7Cjmacdonald%40mtsac.edu%7C0fbc82a2b13749a4222608d928cbfe52%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637585679205067185%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000sdata=KjvijcfrVPGZKGsAn6qX5rMKtulHpmsAzqHEkwz%2B96Y%3Dreserved=0<https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian-Blue=04%7C01%7Cjmacdonald%40mtsac.edu%7C2eeb915058594b1d14b508d929d455c4%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637586814671423758%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000=xKy3v4qvO4spk0zrRxXWZfcsHWxbQC%2FIQ1FbpMB390Y%3D=0> -for-Solvent-Based-Inks.html On the other hand, if the blue colour doesn't interfere with your following staining, you can try to simple make a "double stain". Regards Gudrun -Ursprüngliche Nachricht- Von: Mac Donald, Jennifer via Histonet [mailto:histonet@lists.utsouthwestern.edu] Gesendet: Sonntag, 6. Juni 2021 06:34 An: histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu> Betreff: [Histonet] Prussian Blue Reaction Does anyone know of a way to remove/reduce the Prussian blue reaction? Thanks, Jennifer ___ Histonet mailing list Histonet@lists.utsouthwestern.edu<mailto:Histonet@lists.utsouthwestern.edu> https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonetdata=04%7C01%7Cjmacdonald%40mtsac.edu%7C0fbc82a2b13749a4222608d928cbfe52%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637585679205067185%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000sdata=gRrUmRDEU3BfcA0rEgQOgBvPHIQ05IRM6WozVZiiR1g%3Dreserved=0<https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonet=04%7C01%7Cjmacdonald%40mtsac.edu%7C2eeb915058594b1d14b508d929d455c4%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637586814671433754%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000=1p0zF%2BbxdarMOeh8R%2B2HN3JrGroGYcbuplEXFxO6y1s%3D=0> ___ Histonet mailing list Histonet@lists.utsouthwestern.edu<mailto:Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet<https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonet=04%7C01%7Cjmacdonald%40mtsac.edu%7C2eeb915058594b1d14b508d929d455c4%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637586814671443749%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000=%2FoXTqTKQY18uiQ%2BldVcTbiT
Re: [Histonet] Prussian Blue Reaction
Overstained? Doesn't that mean the tissue contains a lot of iron and you are seeing where it is - which was the reason for doing Prussian blue histochemistry. Gudrun Lang correctly says that mineral acids won't remove it. Oxalic acid is said to dissolve Prussian blue (? by chelation); I've never tried this. If it works, you will no longer see where the iron is. To see features other than the distribution of iron, why not just stain another section from the block with a general-purpose stain like Giemsa or H? John Kiernan London, Canada = = = From: Mac Donald, Jennifer via Histonet Sent: June 7, 2021 12:47 AM To: Gudrun Lang Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Prussian Blue Reaction The tissue was overstained and the blue was interfering with interpretation -Original Message- From: Gudrun Lang Sent: Sunday, June 6, 2021 2:18 AM To: Mac Donald, Jennifer Cc: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Hi Jennifer, Why do you want to reduce the staining? I ask, because the impact of hydrochloric acid on the tissue may influence the following results anyway. I think, that the prussian blue pigment cannot be removed in an easy way. It is resistent to solvents and mineral acids. https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian-Bluedata=04%7C01%7Cjmacdonald%40mtsac.edu%7C0fbc82a2b13749a4222608d928cbfe52%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637585679205067185%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000sdata=KjvijcfrVPGZKGsAn6qX5rMKtulHpmsAzqHEkwz%2B96Y%3Dreserved=0 -for-Solvent-Based-Inks.html On the other hand, if the blue colour doesn't interfere with your following staining, you can try to simple make a "double stain". Regards Gudrun -Ursprüngliche Nachricht- Von: Mac Donald, Jennifer via Histonet [mailto:histonet@lists.utsouthwestern.edu] Gesendet: Sonntag, 6. Juni 2021 06:34 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Prussian Blue Reaction Does anyone know of a way to remove/reduce the Prussian blue reaction? Thanks, Jennifer ___ Histonet mailing list Histonet@lists.utsouthwestern.edu https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonetdata=04%7C01%7Cjmacdonald%40mtsac.edu%7C0fbc82a2b13749a4222608d928cbfe52%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637585679205067185%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000sdata=gRrUmRDEU3BfcA0rEgQOgBvPHIQ05IRM6WozVZiiR1g%3Dreserved=0 ___ 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] Prussian Blue Reaction
The tissue was overstained and the blue was interfering with interpretation -Original Message- From: Gudrun Lang Sent: Sunday, June 6, 2021 2:18 AM To: Mac Donald, Jennifer Cc: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] Prussian Blue Reaction EXTERNAL SENDER- Exercise caution with requests, links, and attachments. Hi Jennifer, Why do you want to reduce the staining? I ask, because the impact of hydrochloric acid on the tissue may influence the following results anyway. I think, that the prussian blue pigment cannot be removed in an easy way. It is resistent to solvents and mineral acids. https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.epsilonpigments.com%2Finorganic-pigment%2Fprussian-blue%2FPrussian-Bluedata=04%7C01%7Cjmacdonald%40mtsac.edu%7C0fbc82a2b13749a4222608d928cbfe52%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637585679205067185%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000sdata=KjvijcfrVPGZKGsAn6qX5rMKtulHpmsAzqHEkwz%2B96Y%3Dreserved=0 -for-Solvent-Based-Inks.html On the other hand, if the blue colour doesn't interfere with your following staining, you can try to simple make a "double stain". Regards Gudrun -Ursprüngliche Nachricht- Von: Mac Donald, Jennifer via Histonet [mailto:histonet@lists.utsouthwestern.edu] Gesendet: Sonntag, 6. Juni 2021 06:34 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Prussian Blue Reaction Does anyone know of a way to remove/reduce the Prussian blue reaction? Thanks, Jennifer ___ Histonet mailing list Histonet@lists.utsouthwestern.edu https://nam12.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.utsouthwestern.edu%2Fmailman%2Flistinfo%2Fhistonetdata=04%7C01%7Cjmacdonald%40mtsac.edu%7C0fbc82a2b13749a4222608d928cbfe52%7Ccc4d4bf20a9e4240aedea7d1d688f935%7C0%7C0%7C637585679205067185%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C2000sdata=gRrUmRDEU3BfcA0rEgQOgBvPHIQ05IRM6WozVZiiR1g%3Dreserved=0 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Prussian Blue Reaction
Hi Jennifer, Why do you want to reduce the staining? I ask, because the impact of hydrochloric acid on the tissue may influence the following results anyway. I think, that the prussian blue pigment cannot be removed in an easy way. It is resistent to solvents and mineral acids. http://www.epsilonpigments.com/inorganic-pigment/prussian-blue/Prussian-Blue -for-Solvent-Based-Inks.html On the other hand, if the blue colour doesn't interfere with your following staining, you can try to simple make a "double stain". Regards Gudrun -Ursprüngliche Nachricht- Von: Mac Donald, Jennifer via Histonet [mailto:histonet@lists.utsouthwestern.edu] Gesendet: Sonntag, 6. Juni 2021 06:34 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Prussian Blue Reaction Does anyone know of a way to remove/reduce the Prussian blue reaction? Thanks, Jennifer ___ 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] Prussian Blue Reaction
Does anyone know of a way to remove/reduce the Prussian blue reaction? Thanks, Jennifer ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
