Tetracycline to bone augmentation

[Javier Adur]

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Dear list,
We want to evaluate the bone augmentation in rabbit tibia using Tetracycline or similar.
Does anyone know a protocol of inmunohistochemistry o reference literature about it.
Thanks in advance.
Javier





                     
         

                  Dr. Javier F. Adur

     Docente/Investigador
- Facultad de Ingeniería - UNER - Argentina

Laboratorio de
Microscopia Aplicada a Estudios Celulares y Moleculares

                            Cátedra Radiaciones
No Ionizantes        
 
                 

[Chris Tully]

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Javier,

I am NOT a Pathologist, but for the last three years have been providing
demonstrations of digital pathology equipment to pathologists across the
world.  I have left that position and opened a consulting company, Image
Incyte, LLC.

I only saw bone samples a few times in my multiple demos.  What I do recall
though is: Errors in animal treatments have been identified by
characteristic tetracycline rings on bones - meaning that you do not have
to specifically stain for tetracycline, it shows up on it's own.  I believe
that the drug is actually incorporated whole in the bone and shows up with
distinctive fluorescence.

I found this note:
http://www.histosearch.com/histonet/Dec02/TetracyclinelabelingofbonA.html
on the Wikipedia page for tetracycline:
http://en.wikipedia.org/wiki/Tetracycline#cite_note-mayton-15

I hope that helps you get started on your search.



Chris Tully
Microscopy and Image Analysis Expert
Image Incyte, LLC
www.ImageIncyte.com
[hidden email]
[hidden email]
240-475-9753 (c)

[image: View my profile on LinkedIn]<http://www.linkedin.com/in/christully/>


On Wed, May 15, 2013 at 10:16 AM, Javier Adur <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear list,
> We want to evaluate the bone augmentation in rabbit tibia using
> Tetracycline or similar.
> Does anyone know a protocol of inmunohistochemistry o reference literature
> about it.
> Thanks in advance.
> Javier
>
>
>
>
>
>
>
>
>                   Dr. Javier F. Adur
>
>      Docente/Investigador
> - Facultad de Ingeniería - UNER - Argentina
>
> Laboratorio de
> Microscopia Aplicada a Estudios Celulares y Moleculares
>
>                             Cátedra Radiaciones
> No Ionizantes
>
>

[Jeremy Adler-4]

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tetracycline fluoresces


Quoting Javier Adur <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear list,
> We want to evaluate the bone augmentation in rabbit tibia using  
> Tetracycline or similar.
> Does anyone know a protocol of inmunohistochemistry o reference  
> literature about it.
> Thanks in advance.
> Javier
>
>
>
>
>
>
>
>
>                   Dr. Javier F. Adur
>
>      Docente/Investigador
> - Facultad de Ingeniería - UNER - Argentina
>
> Laboratorio de
> Microscopia Aplicada a Estudios Celulares y Moleculares
>
>                             Cátedra Radiaciones
> No Ionizantes
>
>

Jeremy Adler
IGP
Rudbeckslaboratoriet
Daghammersköljdsväg 20
751 85 Uppsala
Sweden

0046 (0)18 471 4607

[Doube, Michael]

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Dear Javier,

> We want to evaluate the bone augmentation in rabbit tibia using
> Tetracycline or similar.

I think you are referring to the very old technique of in-vivo labelling
of mineral apposition in calcified tissues, right? In which case you
might consider other labels such as alizarin and calcein (green and
blue). Calcein green in particular gives a stable bright green
fluorescence and works well with fluorescein filters and excitation with
a 488nm laser. Main issue is the yellowish autofluorescence from the
bone matrix and cells themselves. But, that, and the backscattered
reflected light can be used to your advantage to give tissue contrast
without further processing. See e.g. (shameless self-promotion warning):

Doube, M., Firth, E. C. & Boyde, A. Variations in articular calcified
cartilage by site and exercise in the 18-month-old equine distal
metacarpal condyle. Osteoarth Cart 15, 1283–1292 (2007).
http://dx.doi.org/10.1016/j.joca.2007.04.003

And the originals:
1.FROST, H. M. Measurement of human bone formation by means of
tetracycline labelling. Can J Biochem Physiol 41, 31–42 (1963).
2.FROST, H. M. Tetracycline labelling of bone and the zone of
demarcation of osteoid seams. Can J Biochem Physiol 40, 485–489 (1962).

There is lots more literature on labelling protocols, see e.g.
Erben, R. G. & Glösmann, M. Histomorphometry in rodents. Methods Mol.
Biol. 816, 279–303 (2012). http://dx.doi.org/10.1007/978-1-61779-415-5_19


Michael








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[mcammer]

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In the early 1990s I imaged what sounds like this for a renal MD.  He had bone biopsies from patients given tetracycline as an indicator of bone growth which was a diagnostic tool for success/failure or kidney transplant.  The question was whether the confocal could avoid the prep of the standard technique of getting a neat cross section.  But we could not image depth of tetracycline ring accurately by looking at the surface and trying to do a Z series.  And for imaging cross sections, we found the confocal was no improvement over the then clinical practice.  Back then we were using a 60X NA 1.4 Nikon fixed tube length objective; maybe newer microscopes could do better.
Regards,
Michael C.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Jeremy Adler
Sent: Thursday, May 16, 2013 3:28 AM
To: [hidden email]
Subject: Re: Tetracycline to bone augmentation

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tetracycline fluoresces


Quoting Javier Adur <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear list,
> We want to evaluate the bone augmentation in rabbit tibia using
> Tetracycline or similar.
> Does anyone know a protocol of inmunohistochemistry o reference
> literature about it.
> Thanks in advance.
> Javier
>
>
>
>
>
>
>
>
>                   Dr. Javier F. Adur
>
>      Docente/Investigador
> - Facultad de Ingeniería - UNER - Argentina
>
> Laboratorio de
> Microscopia Aplicada a Estudios Celulares y Moleculares
>
>                             Cátedra Radiaciones
> No Ionizantes
>
>

Jeremy Adler
IGP
Rudbeckslaboratoriet
Daghammersköljdsväg 20
751 85 Uppsala
Sweden

0046 (0)18 471 4607

[Tim Feinstein-2]

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Without a doubt you still need sectioning to measure bone deposition by tetracycline fluorescence.  Bone is only 10-30% tissue and at least 70% hydroxyapatite, a mineral that does not transmit light.  This property has given my current research group, which very much would like to see bone cells in their natural environment, more than a little bit of grief.  

cheers,


TF

Timothy Feinstein, PhD
Visiting Research Associate
Laboratory for GPCR Biology
Dept. of Pharmacology & Chemical Biology
University of Pittsburgh, School of Medicine
BST W1301, 200 Lothrop St.
Pittsburgh, PA  15261

On May 16, 2013, at 11:43 AM, Cammer, Michael wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> In the early 1990s I imaged what sounds like this for a renal MD.  He had bone biopsies from patients given tetracycline as an indicator of bone growth which was a diagnostic tool for success/failure or kidney transplant.  The question was whether the confocal could avoid the prep of the standard technique of getting a neat cross section.  But we could not image depth of tetracycline ring accurately by looking at the surface and trying to do a Z series.  And for imaging cross sections, we found the confocal was no improvement over the then clinical practice.  Back then we were using a 60X NA 1.4 Nikon fixed tube length objective; maybe newer microscopes could do better.
> Regards,
> Michael C.
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Jeremy Adler
> Sent: Thursday, May 16, 2013 3:28 AM
> To: [hidden email]
> Subject: Re: Tetracycline to bone augmentation
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> tetracycline fluoresces
>
>
> Quoting Javier Adur <[hidden email]>:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Dear list,
>> We want to evaluate the bone augmentation in rabbit tibia using
>> Tetracycline or similar.
>> Does anyone know a protocol of inmunohistochemistry o reference
>> literature about it.
>> Thanks in advance.
>> Javier
>>
>>
>>
>>
>>
>>
>>
>>
>>                  Dr. Javier F. Adur
>>
>>     Docente/Investigador
>> - Facultad de Ingeniería - UNER - Argentina
>>
>> Laboratorio de
>> Microscopia Aplicada a Estudios Celulares y Moleculares
>>
>>                            Cátedra Radiaciones
>> No Ionizantes
>>
>>
>
>
>
> Jeremy Adler
> IGP
> Rudbeckslaboratoriet
> Daghammersköljdsväg 20
> 751 85 Uppsala
> Sweden
>
> 0046 (0)18 471 4607

[Doube, Michael]

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> Without a doubt you still need sectioning to measure bone deposition
> by tetracycline fluorescence.

Sectioning, or imaging a polished or diamond ultramilled surface of an
arbitrarily thick block of embedded tissue - which is where confocal
becomes especially useful.

Michael


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This message, together with any attachments, is intended for the stated addressee(s) only and may contain privileged or confidential information. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Royal Veterinary College (RVC). If you are not the intended recipient, please notify the sender and be advised that you have received this message in error and that any use, dissemination, forwarding, printing, or copying is strictly prohibited. Unless stated expressly in this email, this email does not create, form part of, or vary any contractual or unilateral obligation. Email communication cannot be guaranteed to be secure or error free as information could be intercepted, corrupted, amended, lost, destroyed, incomplete or contain viruses. Therefore, we do not accept liability for any such matters or their consequences. Communication with us by email will be taken as acceptance of the risks inherent in doing so.