Re: [Histonet] New Bone Stain

[Dr. Michael Gudo (Morphisto GmbH) via Histonet]

Hello Jessica,
Hello Natalia,

we have discussed the question of bone staining methods in our lab and we would 
recommend the following stainings, which will work on resin embedded material:

(1) For an overview and differentiation of various types of bone the 
traditional MASSON-GOLDNER or AZAN staining is quite fine, and should be the 
first try because these stains are more or less easy and fast and the staining 
solutions are not too expensive.

(2) MOVAT-Pentachrome in the original and in the modification after VERHOEFF 
also gives quite good differentiations of resin embedded bones and shows many 
details even in different types of bone. We use these staining here in many 
research projects we do for pharma companies that develop bone replacement 
materials.

(3) van KOSSA is also a very good silver impregnation to show mineralized bone, 
AND there is a possible combination with the MOVAT Pentachrom-Staining.

(4) Alizarinred can also be used to show different gradients of mineralisation; 
the staining kit contains solutions with several pH-values.

(5) Picro-Siriusred and other combinations of picric acid and other stains 
(picro fuchsin, picro polychrom, picro indigocarmin, and 
orcein-picroindicocramin can show good differentiations on tissues and 
cartilage and bone structures.
Remark: Sometimes its a bit difficult to establish picric acid based stainings 
on resin embedded tissues, if the resin has not been completely removed (which 
is possible with technovit 9100 (and some other MMA’s), but not with technovit 
7200).

(6) LEVAI-LACZKO's stain provides a good differentiation between mineralized 
bone matrix, Osteoid and synthetic bone substitutes.

(7) A special „Bone Stain“, published by Villanueva in 2009 
(https://www.tandfonline.com/doi/abs/10.3109/10520297409116928 
<https://www.tandfonline.com/doi/abs/10.3109/10520297409116928>) works best on 
fresh material. We have not yet tested it completely on resin embedded tissues, 
but some of our customers used it successfully.
For this stain, a „staining powder" has to be synthesized from various single 
stains (basic fuchsin, orange G, fastgreen and azure II) and finally mixed into 
a ready to use staining solution; we produce the stain powder and ready to use 
solution here in our lab.

(8) Last but not least HEROVICI’s stain might help beacuse it differentiates 
between new and old collagen structures. Though it was originally developed for 
diagnostics in wound repair, some of our customers use it to differentiate 
between mineralized bone, new bone (Osteoid) and synthetic bone substitutes.

For all staining procedures on resin embedded material its necessary to edge 
the surface with citric acid or hydrogen peroxide, depending on the kind of 
resin you are using. If it is technovit 7200 citric acid should work fine, for 
technovit 9100 you can also try to remove the resin with methoxyethylacetate 
and acetone. Please note, that there can be a significant difference in the 
quality of the stain between Technovit 9100 and Technovit 7200, depending on 
what you want to see. In general, the stain on Technovit 9100 sections will be 
more powerful and brilliant than that on Technovit 7200 sections. 

As mentioned in my previous mail, we have a local dealer in Oklahoma, and if 
you or somebody else is interested in our ready-to-use stainings kits, just let 
me know, I will then make the direct contact.

Kind regards
Michael



> Am 10.07.2018 um 22:15 schrieb Jessica Riggleman via Histonet 
> :
> 
> Hey Jennifer,
> Thanks for your help. I have two questions:
> 
> 1. What is the point of decalcifying if they are in plastic?
> 2. Did you see the counterstain? Right now only the bone is staining. How 
> long did you leave these in the counterstain for?
> Thanks!
> Jessica
> 
> From: Mac Donald, Jennifer [mailto:jmacdon...@mtsac.edu]
> Sent: Tuesday, July 10, 2018 4:07 PM
> To: Jessica Riggleman ; 
> histonet@lists.utsouthwestern.edu
> Subject: Re: [Histonet] New Bone Stain
> 
> I used to do it on plastic sections of in-de-calcified bone biopsies. It’s a 
> silver reaction.
> 
> Get Outlook for iOS
> 
> 
> 
> _
> 
> Jessica Riggleman | Research Associate
> 
> Globus Medical, Inc.
> Valley Forge Business Center
> 2560 General Armistead Avenue | Audubon, PA 19403
> Ph: (610) 930-1800 ext. 2583 | Fax:
> 
> Confidentiality Note:  This email is confidential and intended solely for the 
> use of the individual to whom it is addressed. If you are not the intended 
> recipient, be advised that you have received this email in error and that any 
> use, dissemination, forwarding, printing, or copying of this email is 
> strictly prohibited. If you have received this email in error please contact 
> the sender. Any views or opinions presented are solely those of t

Re: [Histonet] New Bone Stain

[Mac Donald, Jennifer via Histonet]

Auto corrected on my phone. They were not decalcified. The counterstain was a 
brilliant red/fuschia in the osteoid.

Get Outlook for iOS<https://aka.ms/o0ukef>


From: Jessica Riggleman 
Sent: Tuesday, July 10, 2018 1:16 PM
To: Mac Donald, Jennifer; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] New Bone Stain

Hey Jennifer,
Thanks for your help. I have two questions:

1. What is the point of decalcifying if they are in plastic?
2. Did you see the counterstain? Right now only the bone is staining. How long 
did you leave these in the counterstain for?
Thanks!
Jessica

From: Mac Donald, Jennifer [mailto:jmacdon...@mtsac.edu]
Sent: Tuesday, July 10, 2018 4:07 PM
To: Jessica Riggleman ; 
histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] New Bone Stain

I used to do it on plastic sections of in-de-calcified bone biopsies. It’s a 
silver reaction.

Get Outlook for iOS



_

Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:

Confidentiality Note: This email is confidential and intended solely for the 
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From: Jessica Riggleman 
Sent: Tuesday, July 10, 2018 1:01 PM
To: Mac Donald, Jennifer; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] New Bone Stain

Thank You,
Have you ever tried this on thick histology sections? I have a titanium implant 
in this section, so thin is impossible.

Thank You,
Jessica



Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:
An ISO 13485 Registered Company







Confidentiality Note: This email is confidential and intended solely for the 
use of the individual to whom it is addressed. If you are not the intended 
recipient, be advised that you have received this email in error and that any 
use, dissemination, forwarding, printing, or copying of this email is strictly 
prohibited. If you have received this email in error please contact the sender. 
Any views or opinions presented are solely those of the author and do not 
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arising in any way from the receipt or use thereof.
From: Mac Donald, Jennifer [mailto:jmacdon...@mtsac.edu]
Sent: Monday, July 09, 2018 6:09 PM
To: Jessica Riggleman ; 
histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] New Bone Stain

von Kossa will stain the mineralized bone black and a basic fuschin counter 
stain the osteoid will stain deep pink.

Get Outlook for iOS


From: Jessica Riggleman via Histonet 
Sent: Monday, July 9, 2018 1:17 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] New Bone Stain

Hello,
I am trying to assess new bone vs. old bone in an animal model. Any suggestions 
on thick section stains? I have heard Van Gieson's?

Thank You,
Jessica


_

Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:

Confidentiality Note: This email is confidential and intended solely for the 
use of the individual to whom it is addressed. If you are not the intended 
recipient, be advised that you have received this email in error and that any 
use, dissemination, forwarding, printing, or copying of this email is strictly 
prohibited. If you have received this email in error please contact the sender. 
Any views or opinions presented are solely those of the author and do not 
necessarily represent those of Globus Medical, Inc. Although this email and any 
attachments are believed to be free of any virus or other defects which

Re: [Histonet] New Bone Stain

[Jessica Riggleman via Histonet]

Hey Jennifer,
Thanks for your help. I have two questions:

1. What is the point of decalcifying if they are in plastic?
2. Did you see the counterstain? Right now only the bone is staining. How long 
did you leave these in the counterstain for?
Thanks!
Jessica

From: Mac Donald, Jennifer [mailto:jmacdon...@mtsac.edu]
Sent: Tuesday, July 10, 2018 4:07 PM
To: Jessica Riggleman ; 
histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] New Bone Stain

I used to do it on plastic sections of in-de-calcified bone biopsies. It’s a 
silver reaction.

Get Outlook for iOS



_

Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:

Confidentiality Note:  This email is confidential and intended solely for the 
use of the individual to whom it is addressed. If you are not the intended 
recipient, be advised that you have received this email in error and that any 
use, dissemination, forwarding, printing, or copying of this email is strictly 
prohibited. If you have received this email in error please contact the sender. 
Any views or opinions presented are solely those of the author and do not 
necessarily represent those of Globus Medical, Inc. Although this email and any 
attachments are believed to be free of any virus or other defects which might 
affect any computer or IT system into which they are received, no 
responsibility is accepted by Globus Medical, Inc. for any loss or damage 
arising in any way from the receipt or use thereof.



From: Jessica Riggleman 
Sent: Tuesday, July 10, 2018 1:01 PM
To: Mac Donald, Jennifer; histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] New Bone Stain

Thank You,
Have you ever tried this on thick histology sections? I have a titanium implant 
in this section, so thin is impossible.

Thank You,
Jessica



Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:
An ISO 13485 Registered Company







Confidentiality Note:  This email is confidential and intended solely for the 
use of the individual to whom it is addressed. If you are not the intended 
recipient, be advised that you have received this email in error and that any 
use, dissemination, forwarding, printing, or copying of this email is strictly 
prohibited. If you have received this email in error please contact the sender. 
Any views or opinions presented are solely those of the author and do not 
necessarily represent those of Globus Medical, Inc. Although this email and any 
attachments are believed to be free of any virus or other defects which might 
affect any computer or IT system into which they are received, no 
responsibility is accepted by Globus Medical, Inc. for any loss or damage 
arising in any way from the receipt or use thereof.
From: Mac Donald, Jennifer [mailto:jmacdon...@mtsac.edu]
Sent: Monday, July 09, 2018 6:09 PM
To: Jessica Riggleman ; 
histonet@lists.utsouthwestern.edu
Subject: Re: [Histonet] New Bone Stain

von Kossa will stain the mineralized bone black and a basic fuschin counter 
stain the osteoid will stain deep pink.

Get Outlook for iOS


From: Jessica Riggleman via Histonet 
Sent: Monday, July 9, 2018 1:17 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] New Bone Stain

Hello,
I am trying to assess new bone vs. old bone in an animal model. Any suggestions 
on thick section stains? I have heard Van Gieson's?

Thank You,
Jessica


_

Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:

Confidentiality Note: This email is confidential and intended solely for the 
use of the individual to whom it is addressed. If you are not the intended 
recipient, be advised that you have received this email in error and that any 
use, dissemination, forwarding, printing, or copying of this email is strictly 
prohibited. If you have received this email in error please contact the sender. 
Any views or opinions presented are solely those of the author and do not 
necessarily represent those of Globus Medical, Inc. Although this email and any 
attachments are believed to be free of any virus or other defects which might 
affect any computer or IT system into which they are received, no 
responsibility is accepted by Globus Medical, Inc. for any loss or damage 
arising in any way from the receipt or use thereof.


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Re: [Histonet] New Bone Stain

[Dr. Michael Gudo (Morphisto GmbH) via Histonet]

Hello Jessica,

we have a special „bone stain“ staining solution in our portfolio which is 
exactly for the purpose you described. Osteoid will be stained green or red to 
dark red, incomplete mineralized bone bone light red or orange yellow and the 
demarcation zone light green. Its also possible to differentiate lacunae, 
canaliculae, feathered bone, etc. This special stain was developed for 
diagnosis of bone diseases and I think it should be suitable for your purpose.

If you are interested in this staining kit, please let me know, we have a local 
dealer in Oklahoma and so it would be easy to deliver the ready-to-use staining 
solution to you.

Kind regards
Michael



> Am 09.07.2018 um 22:16 schrieb Jessica Riggleman via Histonet 
> :
> 
> Hello,
> I am trying to assess new bone vs. old bone in an animal model. Any 
> suggestions on thick section stains? I have heard Van Gieson's?
> 
> Thank You,
> Jessica
> 
> 
> _
> 
> Jessica Riggleman | Research Associate
> 
> Globus Medical, Inc.
> Valley Forge Business Center
> 2560 General Armistead Avenue | Audubon, PA 19403
> Ph: (610) 930-1800 ext. 2583 | Fax:
> 
> Confidentiality Note:  This email is confidential and intended solely for the 
> use of the individual to whom it is addressed. If you are not the intended 
> recipient, be advised that you have received this email in error and that any 
> use, dissemination, forwarding, printing, or copying of this email is 
> strictly prohibited. If you have received this email in error please contact 
> the sender. Any views or opinions presented are solely those of the author 
> and do not necessarily represent those of Globus Medical, Inc. Although this 
> email and any attachments are believed to be free of any virus or other 
> defects which might affect any computer or IT system into which they are 
> received, no responsibility is accepted by Globus Medical, Inc. for any loss 
> or damage arising in any way from the receipt or use thereof.
> 
> 
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Re: [Histonet] New Bone Stain

[Mac Donald, Jennifer via Histonet]

von Kossa will stain the mineralized bone black and a basic fuschin counter 
stain the osteoid will stain deep pink.

Get Outlook for iOS<https://aka.ms/o0ukef>


From: Jessica Riggleman via Histonet 
Sent: Monday, July 9, 2018 1:17 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] New Bone Stain

Hello,
I am trying to assess new bone vs. old bone in an animal model. Any suggestions 
on thick section stains? I have heard Van Gieson's?

Thank You,
Jessica


_

Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:

Confidentiality Note: This email is confidential and intended solely for the 
use of the individual to whom it is addressed. If you are not the intended 
recipient, be advised that you have received this email in error and that any 
use, dissemination, forwarding, printing, or copying of this email is strictly 
prohibited. If you have received this email in error please contact the sender. 
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attachments are believed to be free of any virus or other defects which might 
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responsibility is accepted by Globus Medical, Inc. for any loss or damage 
arising in any way from the receipt or use thereof.


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[Histonet] New Bone Stain

[Jessica Riggleman via Histonet]

Hello,
I am trying to assess new bone vs. old bone in an animal model. Any suggestions 
on thick section stains? I have heard Van Gieson's?

Thank You,
Jessica


_

Jessica Riggleman | Research Associate

Globus Medical, Inc.
Valley Forge Business Center
2560 General Armistead Avenue | Audubon, PA 19403
Ph: (610) 930-1800 ext. 2583 | Fax:

Confidentiality Note:  This email is confidential and intended solely for the 
use of the individual to whom it is addressed. If you are not the intended 
recipient, be advised that you have received this email in error and that any 
use, dissemination, forwarding, printing, or copying of this email is strictly 
prohibited. If you have received this email in error please contact the sender. 
Any views or opinions presented are solely those of the author and do not 
necessarily represent those of Globus Medical, Inc. Although this email and any 
attachments are believed to be free of any virus or other defects which might 
affect any computer or IT system into which they are received, no 
responsibility is accepted by Globus Medical, Inc. for any loss or damage 
arising in any way from the receipt or use thereof.


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