[Histonet] Bone processing help

Mon Mar 18 08:17:03 CDT 2013

Dr. De la Vega,

My name is Jack Ratliff and I Chair the Hard Tissue Committee for the
National Society for Histotechnology. I am not sure if anyone has provided
you with a response to your message, but maybe I can be of assistance. Just
so you know, the NSH prides itself on being an educational resource for
those working in the field of Histology. The Hard Tissue Committee
represents the educational arm of the society for those working with "hard
tissues" like bone, biomaterials and medical device implants and especially
those type of histological specimens embedded into resins or plastics!

I would like to first direct your attention to the NSH website at
www.nsh.org. If you go to the following link:
http://www.nsh.org/content/benefits-membership, you will find information
concerning membership benefits and how to join so that you can take
advantage of what the society can do for you and your laboratory! Please
visit this site first and then feel free to get back to me with any
questions.

As for your current issues, I can definitely help you with all that you are
experiencing. At the moment I am currently away from the US and in Germany
until the 26th of March working with a group (Rowiak GmbH - www.rowiak.de)
that has developed a non-contact laser microtome (Tissue
Surgeon<http://www.rowiak.de/index.php?id=19>)
that can do what you are doing now and produce stained slides @ 10-15
microns in 30 minutes from a resin/plastic embedded block! If you can give
me a few more hours, I will personally respond to your message privately.
In the meantime, please visit the NSH website (www.nsh.org), consider its
membership and also look to join the Hard Tissue
Committee<http://www.nsh.org/committee-detail/1044> so
that I can provide you with meeting updates.

With that said and seeing that you are in Massachusetts, on May the 4th,
2013, Polysciences, Inc. is hosting a full day educational event focusing
on the *Histological Applications & Techniques for Bone, Biomaterials and
Medical Device Implants*. I will also be a speaker at this event. Those
that attend can expect to further their knowledge, understanding and
training of specialized histology techniques associated with bone,
biomaterials and medical device implant specimen types and attendees will
learn of the applicational relevance of the techniques used in the
evaluation safety and efficacy of therapeutic treatments. Registration is
currently open with an *Early Bird registration set to end this Friday,
March 22nd*. The National Society for Histotechnology (NSH) will be
providing 6 CEUs and Polysciences, Inc. will be providing a discount to any
current NSH member as outlined below:

*WORKSHOP FEES*
*
*
*Before March 22, 2013:  $149.00 for NSH Members / $199.00 for Non-Members*
*
*
*After March 22, 2013:  $179.00 for NSH Members / $229.00 for Non-Members*

For the complete details of this full day *Histological Applications &
Techniques for Bone, Biomaterials and Medical Device Implants *event and to
register online, please visit the following link: *
http://www.polysciences.com/Interactive-Histology-Forum-About/185/* and
sign up today to participate in the discussion of these specialized
histology specimen applications and techniques that are rarely shared or
even discussed on Histonet! You will not want to miss out on the
information presented by 4 expert speakers in the field, all course
materials, meals and a complete program book also containing technique
specific protocols that you can repeat back in your lab!

Best Regards,

Jack

Jack L. Ratliff
Owner/Histologist, Ratliff Histology Consultants, LLC
Chairman, Hard Tissue Committee - National Society for Histotechnology

389 Nichol Mill Lane
Franklin, TN 37067
(317) 281-1975 (c)
(615) 236-4901 (o)
(615) 236-4962 (f)
jratliff <@t> ratliffhistology.com

On Sun, Mar 17, 2013 at 8:09 PM, Jack Ratliff <ratliffjack <@t> hotmail.com>wrote:

>
>
>
> Begin forwarded message:
>
> *From:* "De La Vega Amador, Rodolfo Enrique" <RDELAVEGAAMADOR <@t> PARTNERS.ORG
> >
> *Date:* March 15, 2013, 9:41:23 PM GMT+01:00
> *To:* "histonet <@t> lists.utsouthwestern.edu" <
> histonet <@t> lists.utsouthwestern.edu>
> *Subject:* *[Histonet] Bone processing help*
>
> Hi everybody,
>
> I am fairly new to all Histotechnology processes. We mainly work in our
> lab with mineralized bone with implants, so we fix them, dehydrate them,
> embed them in MMA, cut them, glued them to slides, ground them manually and
> then stain them. I need help in some parts of the process that need
> improvement:
>
>
>  1.  We use Loctite 4471 on the samples, put them under vacuum, apply them
> to the slide and more vacuum. There's good results but there are some
> bubbles that still appear. Suggestions?
>  2.  I can't seem to get the yellow/orange on bone with the Van Gieson
> stain. I've been doinga preheat at 55 °C, etching, rinse in DI water,
> Sanderson´s Rapid Bone Stain, running tap water, Van Gieson (commercial
> from DHM) for 30 seconds to 5 minutes, paper dry and quickly dehydrate with
> 100% EtOH. What am I doing wrong?
>  3.  The stain's been running when I add the glue to cover slip the
> slides. Especially the green color. Why does this happen?
>  4.  Cover slipping is done with cyano acrylate, under suction as before,
> but bubbles still appear. I've tried placing the cover slip on an angle and
> wait for the glue to evenly spread, but the bone has just too many pores
> and air bubbles appear. Please help.
>
> I greatly appreciate any help on the subject.
>
> Best,
>
> Rodolfo De la Vega, MD.
> Research Fellow
> Laboratory for Musculoskeletal Research and Innovation
> Massachusetts General Hospital - Jackson Building 1120
>
>
>
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