[Histonet] RE: gout crystals

HOOD MS. GLENDA HOOD <@t> tarleton.edu
Wed Jul 7 11:39:05 CDT 2010


It is true that gout crystals are water-soluble, and we are all taught that... but a few years ago there was a published article (JOH??) that showed that many crystals DO survive after water-based fixation. Since you already have the specimen in formalin, at least try processing and see if they can be demonstrated. Couldn't hurt, not nearly as much as the patient would for a re-biopsy!

Glenda F. Hood, M.Ed., HT(ASCP)
Instructor and Program Director
Histotechnology Program
Tarleton State University
817-926-1101 x6



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Wednesday, July 07, 2010 11:22 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 80, Issue 7

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Today's Topics:

   1. Rnase free slides? (Caroline Bass)
   2. Gout Crytals (Behnaz Sohrab)
   3. RE: Gout Crytals (Weems, Joyce)
   4. Re: Gout Crytals (Jennifer MacDonald)
   5. RE: Gout Crytals (sgoebel <@t> xbiotech.com)
   6. Schmitz, Sandy is out of the office.
      (Sandy.Schmitz <@t> leica-microsystems.com)
   7. RE: Gout Crytals (Joyce Cline)
   8. Re: bat wing histology (Mohit Chadha)
   9. RE: Gout Crytals (Douglas,Joseph)
  10. used histology equipment (dcojita <@t> tampabay.rr.com)
  11. Re: used histology equipment (Drew Meyer)
  12. Re: Rnase free slides? (Emily Sours)
  13. Re: bat wing histology (Amos Brooks)
  14. Has anyone used a biotin block in their antibody diluent?
      (Jennifer Campbell)
  15. Biotin block (Jim Reilly)
  16. RE: Has anyone used a biotin block in their antibody      diluent?
      (Mauger, Joanne)
  17. Re: used histology equipment (Kim.Donadio <@t> bhcpns.org)
  18. RE: used histology equipment (Douglas,Joseph)
  19. RE: used histology equipment (Douglas,Joseph)
  20. Reprocess (sgoebel <@t> xbiotech.com)
  21. Re: Reprocess (Catherine Simonson)
  22. RE: Reprocess (Cheri Miller)
  23. RE: Reprocess (Cheri Miller)
  24. RE: Reprocess (Mahoney,Janice A)


----------------------------------------------------------------------

Message: 1
Date: Tue, 06 Jul 2010 15:49:58 -0400
From: Caroline Bass <cbass <@t> wfubmc.edu>
Subject: [Histonet] Rnase free slides?
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <C8590126.2888%cbass <@t> wfubmc.edu>
Content-Type: text/plain;       charset="US-ASCII"

Hello,

I'm doing RNA work for the first time. My plan is to take a fresh rat brain,
block quickly, freeze by immersing in dry-ice cooled isopentane, storing at
-80, collecting tissue sections (thickness will be determined, somewhere
between 20 and 300 um), and punching the particular regions I need out of
the sections. I will then isolate RNA from the punches for qPCR analysis.

Questions:

1) does this sound like a viable plan?
2) and suggestions, what to be careful of?
3) where do I have to be careful of Rnase, should I use disposable blades,
cleaned with Rnase away?
4) where can I find Rnase free slides, or should I just make my own. I
usually use charged slides.

Any and all suggestions will be appreciated. I'm new to this and don't know
where I will have problems.

Thanks!

Caroline




------------------------------

Message: 2
Date: Tue, 06 Jul 2010 13:12:49 -0700
From: "Behnaz Sohrab" <SohrabB1 <@t> ah.org>
Subject: [Histonet] Gout Crytals
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4C332BD0.4347.0054.1 <@t> ah.org>
Content-Type: text/plain; charset=US-ASCII

Please refresh my memory!! Processing For Gout Crystal !1 Do I skip all alcohols? Tissue has been fixed in formalin.?
Thank you


------------------------------

Message: 3
Date: Tue, 6 Jul 2010 16:35:41 -0400
From: "Weems, Joyce" <JWeems <@t> sjha.org>
Subject: RE: [Histonet] Gout Crytals
To: Behnaz Sohrab <SohrabB1 <@t> ah.org>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <92AD9B20A6C38C4587A9FEBE3A30E164015E968425 <@t> CHEXCMS10.one.ads.che.org>
Content-Type: text/plain; charset="us-ascii"

Should skip the formalin!!! They are water soluable... :>(

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Behnaz Sohrab
Sent: Tuesday, July 06, 2010 16:13
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Gout Crytals

Please refresh my memory!! Processing For Gout Crystal !1 Do I skip all alcohols? Tissue has been fixed in formalin.?
Thank you
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------------------------------

Message: 4
Date: Tue, 6 Jul 2010 13:50:19 -0700
From: Jennifer MacDonald <JMacDonald <@t> mtsac.edu>
Subject: Re: [Histonet] Gout Crytals
To: "Behnaz Sohrab" <SohrabB1 <@t> ah.org>
Cc: histonet <@t> lists.utsouthwestern.edu,
        histonet-bounces <@t> lists.utsouthwestern.edu
Message-ID:
        <OF6246D4BB.19FA5577-ON88257758.0072608E-88257758.00727EDE <@t> mtsac.edu>
Content-Type: text/plain; charset="US-ASCII"

Uric acid crystals are water soluble.  Avoid formalin and fix in absolute
alcohol and start the processing of the tissue in absolute alcohol.

Jennifer




"Behnaz Sohrab" <SohrabB1 <@t> ah.org>
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
07/06/2010 01:28 PM

To
<histonet <@t> lists.utsouthwestern.edu>
cc

Subject
[Histonet] Gout Crytals






Please refresh my memory!! Processing For Gout Crystal !1 Do I skip all
alcohols? Tissue has been fixed in formalin.?
Thank you
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



------------------------------

Message: 5
Date: Tue, 06 Jul 2010 13:55:24 -0700
From: sgoebel <@t> xbiotech.com
Subject: RE: [Histonet] Gout Crytals
To: "Behnaz Sohrab" <SohrabB1 <@t> ah.org>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <20100706135524.9e2d9aa830e8449a2412eb1e4f2f067e.8ce03ce3da.wbe <@t> email04.secureserver.net>

Content-Type: text/plain; charset="utf-8"


   If  you  have  fixed  in formalin the gout crystals are all gone!!!!&   nbsp;  Have to start over with a new sample if possible.  The crystals


   Sarah Goebel, B.A.
   Histotechnician

   XBiotech USA Inc.
   8201 East Riverside Dr. Bldg 4 Suite 100

   Austin,
   (512)386-5107

   -------- Original Message --------
   Subject: [Histonet] Gout Crytals
   From: "Behnaz Sohrab" <[1]SohrabB1 <@t> ah.   Date: Tue, July 06, 2010 1:12 pm
   To: <[2]histonet <@t> lis   Please  refresh  my  memory!! Processing For Gout Crystal !1 Do I skip
   all alc   Thank you
   _______________________________________________
   Histonet mailing list
   [3]Histonet <@t> lists.utsou   [4]http:
References

   1. 3D"mailto://SohrabB1@ah.org"/
   2. 3D"mailto://histonet@lists.utsouthwestern.edu"/
   3. 3D"mailto://Histonet@lists.utsouthwestern.edu"/
   4. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet"


------------------------------

Message: 6
Date: Tue, 6 Jul 2010 16:00:56 -0500
From: Sandy.Schmitz <@t> leica-microsystems.com
Subject: [Histonet] Schmitz, Sandy is out of the office.
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <OF180DFE40.BFC56A43-ON86257758.00737178-86257758.00737178 <@t> leica-microsystems.com>

Content-Type: text/plain; charset=US-ASCII


I will be out of the office starting  07/05/2010 and will not return until
07/07/2010.



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For more information please visit http://www.messagelabs.com/email
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------------------------------

Message: 7
Date: Tue, 6 Jul 2010 16:56:52 -0400
From: Joyce Cline <Joyce.Cline <@t> wchsys.org>
Subject: RE: [Histonet] Gout Crytals
To: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <FE2197D76184F4408CFEBD95C28282685BF47AFF8A <@t> WCHSXCHCM.wchsys.org>
Content-Type: text/plain; charset="us-ascii"

This works for us.
Process normally, we usually fix in 100% alcohol.
Cut slide
Formula 83 or Xylene 20 seconds
Form 83 or Xylene/100% alcohol mixed 50/50 for 20 seconds
Eosin for 20 seconds
100% alcohol 20 seconds
100% alcohol 20 seconds
Form 83 or Xylene/100% alcohol mixed 50/50 for 20 seconds
Form 83 or Xylene for 20 seconds
coverslip normally

Joyce Cline, Technical Specialist
Hagerstown Medical Laboratory
301-665-4980
fax 301-665-4941
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Behnaz Sohrab [SohrabB1 <@t> ah.org]
Sent: Tuesday, July 06, 2010 4:12 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Gout Crytals

Please refresh my memory!! Processing For Gout Crystal !1 Do I skip all alcohols? Tissue has been fixed in formalin.?
Thank you
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

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------------------------------

Message: 8
Date: Tue, 6 Jul 2010 17:10:17 -0400
From: Mohit Chadha <mchadha1 <@t> gmail.com>
Subject: Re: [Histonet] bat wing histology
To: Amos Brooks <amosbrooks <@t> gmail.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <AANLkTin1AF9XdxXXXT0ikaHoHEnZsykcu10IZZPMBPfZ <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Thank you everyone for replying, much appreciated.

Having also talked to people in my dept, I have a rudimentary protocol
ready. Of course, I will have to tweak it to see what works. I will be using
anti PGP9.5 antibody for neuronal immunology.

I am still not sure how to section the wing. In most likelihood, I will be
using freezing sliding microtome. The "swiss roll" method sound good and I
will definitely try it. I am thinking that since the wing membrane is thin
(~30 um), I will also try to use the whole mount of small pieces.

Any other thoughts and advice would be appreciated.

Thank you,
Mohit Chadha,
Univ of Maryland.






On Fri, Jul 2, 2010 at 4:14 PM, Amos Brooks <amosbrooks <@t> gmail.com> wrote:

> Hi,
>     I do hope you are looking at cross sections of the wing and not the
> flat. That would be very difficult indeed. For good cross sections I would
> try a "Swiss Roll". This is a way of demonstrating a large amount of cross
> sectional area in small space. Take the membrane and fix it by submersion in
> the fixative of your choice. Prior to processing roll the whole membrane up
> then cut the membrane log into sections small enough to fit in a cassette.
> You can use foam biopsy pads to support this shape. Embed it and section it
> on edge to show a long coiled membrane. The hairs should be able to be
> displayed in this way as well. To show a lot of membrane at the same time
> you could place multiple rolls in one cassette. This should work well.
>
> Good Luck,
> Amos
>
>
> Message: 21
> Date: Thu, 1 Jul 2010 11:46:17 -0400
> From: Mohit Chadha <mchadha1 <@t> gmail.com>
> Subject: [Histonet] bat wing histology
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
>        <AANLkTikKGXt1_uY5qkdVhP04fTyIUOCWY9XNVBlEQgW4 <@t> mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> Hello everyone,
>
> This is my very first post and I am desperately looking for help. I am new
> to histology, so any help would be much appreciated.
>
> I am studying the peripheral sensory innervation of bat wings. As a first
> step, I would like to demonstrate the innervation pattern on the different
> parts of the wing membrane (a whole mount of the wing?). Second, I would
> like to demonstrate the mechanoreceptor make-up of the tiny hairs on the
> wing membrane.
>
> Bat wings are highly elastic, with numerous folds, a thickness of about
> 35-45 microns (in the species I study), a network of thin collagen bundles,
> and pigmented superficial epidermal layers.
> I could provide more information if required.
>
> Hoping to hear back from the members.
> Thank you.
>


------------------------------

Message: 9
Date: Tue, 6 Jul 2010 16:22:46 -0500
From: "Douglas,Joseph" <jdouglas <@t> mdanderson.org>
Subject: RE: [Histonet] Gout Crytals
To: 'Jennifer MacDonald' <JMacDonald <@t> mtsac.edu>, Behnaz Sohrab
        <SohrabB1 <@t> ah.org>
Cc: "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>,
        "histonet-bounces <@t> lists.utsouthwestern.edu"
        <histonet-bounces <@t> lists.utsouthwestern.edu>
Message-ID:
        <DACC27F195E63D44BDF7ADC5C52051AF1F02004ABE <@t> DCPWVMBXC0VS1.mdanderson.edu>

Content-Type: text/plain; charset="us-ascii"

REMOVE

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jennifer MacDonald
Sent: Tuesday, July 06, 2010 3:50 PM
To: Behnaz Sohrab
Cc: histonet <@t> lists.utsouthwestern.edu; histonet-bounces <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Gout Crytals

Uric acid crystals are water soluble.  Avoid formalin and fix in absolute
alcohol and start the processing of the tissue in absolute alcohol.

Jennifer




"Behnaz Sohrab" <SohrabB1 <@t> ah.org>
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
07/06/2010 01:28 PM

To
<histonet <@t> lists.utsouthwestern.edu>
cc

Subject
[Histonet] Gout Crytals






Please refresh my memory!! Processing For Gout Crystal !1 Do I skip all
alcohols? Tissue has been fixed in formalin.?
Thank you
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



------------------------------

Message: 10
Date: Tue, 6 Jul 2010 18:29:09 -0400
From: <dcojita <@t> tampabay.rr.com>
Subject: [Histonet] used histology equipment
To: "'histonet'" <Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <!~!UENERkVCMDkAAQACAAAAAAAAAAAAAAAAABgAAAAAAAAAdq7zShuC4ki+F530qNQCZMKAAAAQAAAAgjEUdn4MYUeuJgf6zEQgOAEAAAAA <@t> tampabay.rr.com>

Content-Type: text/plain;       charset="us-ascii"

Does anyone know of a reputable dealer for used equipment?



------------------------------

Message: 11
Date: Tue, 6 Jul 2010 18:55:53 -0400
From: Drew Meyer <41dmb41 <@t> gmail.com>
Subject: Re: [Histonet] used histology equipment
To: dcojita <@t> tampabay.rr.com
Cc: histonet <Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <AANLkTikUIgjmqaDqwBMgz-yJQJ4IymZoL01c83KRZ_DR <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Absolutely... Southeast Pathology Instrument Service out of Charleston, SC.
The owner's name is Michael Dietrich.  I've done business with him before
and they are great people, very honest and they stand behind their
instruments.  I would highly recommend them to anyone.  Contact Michael
directly and tell him Drew Meyer from Atlanta referred you.

http://southeastpathology.com/

Drew

On Tue, Jul 6, 2010 at 18:29, <dcojita <@t> tampabay.rr.com> wrote:

> Does anyone know of a reputable dealer for used equipment?
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


------------------------------

Message: 12
Date: Tue, 6 Jul 2010 19:24:44 -0400
From: Emily Sours <talulahgosh <@t> gmail.com>
Subject: Re: [Histonet] Rnase free slides?
To: Caroline Bass <cbass <@t> wfubmc.edu>,
        histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <AANLkTimZmVTAV6GDp_QN-QDzmY8a9DR8Bdm4aqBxoHO3 <@t> mail.gmail.com>
Content-Type: text/plain; charset=UTF-8

Honestly, I think RNases are a bunch of hooha.  If you're being
careful anyway because you're doing a PCR, that should be enough.
Wear gloves, be sterile.
When I worked with a Russian post-doc, she said she did RNA in situ
hybridization without gloves and it worked.  Of course, god only knows
what her protocol was, as she had some crazy stories about Russian
labs.

Emily

--
Dark Pictures, thrones and stones that pilgrims kiss
And poems that take a thousand years to die
But ape the immortality of this
Red label on a little butterfly.

-Vladimir Nabokov, concluding stanza of ???A Discovery??? 1941.



On Tue, Jul 6, 2010 at 3:49 PM, Caroline Bass <cbass <@t> wfubmc.edu> wrote:
> Hello,
>
> I'm doing RNA work for the first time. My plan is to take a fresh rat brain,
> block quickly, freeze by immersing in dry-ice cooled isopentane, storing at
> -80, collecting tissue sections (thickness will be determined, somewhere
> between 20 and 300 um), and punching the particular regions I need out of
> the sections. I will then isolate RNA from the punches for qPCR analysis.
>
> Questions:
>
> 1) does this sound like a viable plan?
> 2) and suggestions, what to be careful of?
> 3) where do I have to be careful of Rnase, should I use disposable blades,
> cleaned with Rnase away?
> 4) where can I find Rnase free slides, or should I just make my own. I
> usually use charged slides.
>
> Any and all suggestions will be appreciated. I'm new to this and don't know
> where I will have problems.
>
> Thanks!
>
> Caroline
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>



------------------------------

Message: 13
Date: Tue, 6 Jul 2010 22:10:44 -0400
From: Amos Brooks <amosbrooks <@t> gmail.com>
Subject: Re: [Histonet] bat wing histology
To: Mohit Chadha <mchadha1 <@t> gmail.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <AANLkTil_36VIlv0rQs551J8WkwwQjBZ3hYLwPbJ2FQxy <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Hi,
     We did this on derm punch biopsies for a while. We used PLP
(paraformaldehyde-lysine periodate) fixative. This was also done on a
sliding microtome at 50um with piles of dry ice to keep the sucrose frozen.
It was a bit of a pain in the butt, but we got decent results. I think the
sectioning process could have been easier, but the PI was obsessed with not
modifying the project at all, even if it meant improvements.
     We stained these as floating sections in 96 well plates, so in this
case I would be concerned that the wing rolls would un-roll if you were to
do the swiss roll thing I described.

Have fun :-)
Amos

On Tue, Jul 6, 2010 at 5:10 PM, Mohit Chadha <mchadha1 <@t> gmail.com> wrote:

> Thank you everyone for replying, much appreciated.
>
> Having also talked to people in my dept, I have a rudimentary protocol
> ready. Of course, I will have to tweak it to see what works. I will be using
> anti PGP9.5 antibody for neuronal immunology.
>
> I am still not sure how to section the wing. In most likelihood, I will be
> using freezing sliding microtome. The "swiss roll" method sound good and I
> will definitely try it. I am thinking that since the wing membrane is thin
> (~30 um), I will also try to use the whole mount of small pieces.
>
> Any other thoughts and advice would be appreciated.
>
> Thank you,
> Mohit Chadha,
> Univ of Maryland.
>
>
>
>
>
>
>
> On Fri, Jul 2, 2010 at 4:14 PM, Amos Brooks <amosbrooks <@t> gmail.com> wrote:
>
>> Hi,
>>     I do hope you are looking at cross sections of the wing and not the
>> flat. That would be very difficult indeed. For good cross sections I would
>> try a "Swiss Roll". This is a way of demonstrating a large amount of cross
>> sectional area in small space. Take the membrane and fix it by submersion in
>> the fixative of your choice. Prior to processing roll the whole membrane up
>> then cut the membrane log into sections small enough to fit in a cassette.
>> You can use foam biopsy pads to support this shape. Embed it and section it
>> on edge to show a long coiled membrane. The hairs should be able to be
>> displayed in this way as well. To show a lot of membrane at the same time
>> you could place multiple rolls in one cassette. This should work well.
>>
>> Good Luck,
>> Amos
>>
>>
>> Message: 21
>> Date: Thu, 1 Jul 2010 11:46:17 -0400
>> From: Mohit Chadha <mchadha1 <@t> gmail.com>
>> Subject: [Histonet] bat wing histology
>> To: histonet <@t> lists.utsouthwestern.edu
>> Message-ID:
>>        <AANLkTikKGXt1_uY5qkdVhP04fTyIUOCWY9XNVBlEQgW4 <@t> mail.gmail.com>
>> Content-Type: text/plain; charset=ISO-8859-1
>>
>> Hello everyone,
>>
>> This is my very first post and I am desperately looking for help. I am new
>> to histology, so any help would be much appreciated.
>>
>> I am studying the peripheral sensory innervation of bat wings. As a first
>> step, I would like to demonstrate the innervation pattern on the different
>> parts of the wing membrane (a whole mount of the wing?). Second, I would
>> like to demonstrate the mechanoreceptor make-up of the tiny hairs on the
>> wing membrane.
>>
>> Bat wings are highly elastic, with numerous folds, a thickness of about
>> 35-45 microns (in the species I study), a network of thin collagen
>> bundles,
>> and pigmented superficial epidermal layers.
>> I could provide more information if required.
>>
>> Hoping to hear back from the members.
>> Thank you.
>>
>
>


------------------------------

Message: 14
Date: Tue, 6 Jul 2010 19:11:14 -0700
From: "Jennifer Campbell" <jcampbell <@t> vdxpathology.com>
Subject: [Histonet] Has anyone used a biotin block in their antibody
        diluent?
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <5658CBDB9EAE6545ABE50D2563D81BF8181FE1 <@t> VDXSERVER01.vdxpathology.local>

Content-Type: text/plain;       charset="iso-8859-1"

Hi All,

 Has anyone ever used a Biotin block in their primary anitbody diluent?  I have been having problems with nonspecific staining, which I suspect is due to endogenous biotin.  I plan on decreasing my antigen retrieval time, as someone has told me that an antigen retrieval that is too vigorous may cause the unmasking of biotin, and its subsequent staining. I would like to know if anyone has had any luck using a biotin block in their diluent because I may try that as well.

Thanks,

Jennifer Campbell


------------------------------

Message: 15
Date: Wed, 7 Jul 2010 09:05:17 +0100
From: "Jim Reilly" <j.h.reilly <@t> clinmed.gla.ac.uk>
Subject: [Histonet] Biotin block
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <FCD2109433379347B248142CAE948EDFCC8D86 <@t> exchange-be6.centre.ad.gla.ac.uk>

Content-Type: text/plain;       charset="us-ascii"

Hello Jennifer

I use the Avidin/Biotin blocking kit from Vector SP-2001  I mix the
Avidin D with my normal blocking serum and the biotin I add to my
primary antibody diluent. This seems to work well for most tissue types.

Cheers

Jim



------------------------------

Message: 16
Date: Wed, 7 Jul 2010 07:32:52 -0400
From: "Mauger, Joanne" <MAUGER <@t> email.chop.edu>
Subject: [Histonet] RE: Has anyone used a biotin block in their
        antibody        diluent?
To: Jennifer Campbell <jcampbell <@t> vdxpathology.com>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <443F5B475A9BF647AB962E834884EBAD278D1277FE <@t> EX7CCRPW03V1.chop.edu>
Content-Type: text/plain; charset="us-ascii"

Jennifer,
Vector has an avidin-biotin blocking kit that works well by itself or mixed with reagents- very reasonable cost.
Jo

Joanne Mauger HT(ASCP)QIHC
Children's Hospital of Philadelphia
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jennifer Campbell [jcampbell <@t> vdxpathology.com]
Sent: Tuesday, July 06, 2010 10:11 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Has anyone used a biotin block in their antibody diluent?

Hi All,

 Has anyone ever used a Biotin block in their primary anitbody diluent?  I have been having problems with nonspecific staining, which I suspect is due to endogenous biotin.  I plan on decreasing my antigen retrieval time, as someone has told me that an antigen retrieval that is too vigorous may cause the unmasking of biotin, and its subsequent staining. I would like to know if anyone has had any luck using a biotin block in their diluent because I may try that as well.

Thanks,

Jennifer Campbell
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



------------------------------

Message: 17
Date: Wed, 7 Jul 2010 08:37:27 -0500
From: Kim.Donadio <@t> bhcpns.org
Subject: Re: [Histonet] used histology equipment
To: <dcojita <@t> tampabay.rr.com>
Cc: 'histonet' <Histonet <@t> lists.utsouthwestern.edu>,
        histonet-bounces <@t> lists.utsouthwestern.edu
Message-ID:
        <OF32BE6EED.DA4838B0-ON86257759.004AAB55-86257759.004AD74A <@t> bhcpns.org>
Content-Type: text/plain;       charset="US-ASCII"

Hi Diane,
                    Because of your location I would recommend
Micro-optics of Florida. Mike Jones or Tom Christy. The number is
954-791-0082. They are great people to work with and I have found them
very reasonable on their prices.

Hope this helps!



Kim Donadio
Pathology Supervisor
Baptist Hospital
1000 W Moreno St.
Pensacola FL 32501
Phone (850) 469-7718
Fax (850) 434-4996



<dcojita <@t> tampabay.rr.com>
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
07/06/2010 05:29 PM

To
"'histonet'" <Histonet <@t> lists.utsouthwestern.edu>
cc

Subject
[Histonet] used histology equipment






Does anyone know of a reputable dealer for used equipment?

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



-----------------------------------------
All electronic data transmissions originating from or sent to
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This message along with any attached data, are the confidential and
proprietary communications of BHC and are intended to be received
only by the individual or individuals to whom the message has been
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strictly prohibited and may violate State or Federal Law. If you
have received this transmission in error, please delete or destroy
all copies of this message.  For questions, contact the BHC Privacy
Officer at (850) 434-4472.  Rev.10/07.

------------------------------

Message: 18
Date: Wed, 7 Jul 2010 08:40:37 -0500
From: "Douglas,Joseph" <jdouglas <@t> mdanderson.org>
Subject: RE: [Histonet] used histology equipment
To: 'Drew Meyer' <41dmb41 <@t> gmail.com>, "dcojita <@t> tampabay.rr.com"
        <dcojita <@t> tampabay.rr.com>
Cc: histonet <Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <DACC27F195E63D44BDF7ADC5C52051AF1F02004ABF <@t> DCPWVMBXC0VS1.mdanderson.edu>

Content-Type: text/plain; charset="us-ascii"

REMOVE FROM DATABASE

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Drew Meyer
Sent: Tuesday, July 06, 2010 5:56 PM
To: dcojita <@t> tampabay.rr.com
Cc: histonet
Subject: Re: [Histonet] used histology equipment

Absolutely... Southeast Pathology Instrument Service out of Charleston, SC.
The owner's name is Michael Dietrich.  I've done business with him before
and they are great people, very honest and they stand behind their
instruments.  I would highly recommend them to anyone.  Contact Michael
directly and tell him Drew Meyer from Atlanta referred you.

http://southeastpathology.com/

Drew

On Tue, Jul 6, 2010 at 18:29, <dcojita <@t> tampabay.rr.com> wrote:

> Does anyone know of a reputable dealer for used equipment?
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



------------------------------

Message: 19
Date: Wed, 7 Jul 2010 08:41:59 -0500
From: "Douglas,Joseph" <jdouglas <@t> mdanderson.org>
Subject: RE: [Histonet] used histology equipment
To: "'Kim.Donadio <@t> bhcpns.org'" <Kim.Donadio <@t> bhcpns.org>,
        "dcojita <@t> tampabay.rr.com" <dcojita <@t> tampabay.rr.com>
Cc: 'histonet' <Histonet <@t> lists.utsouthwestern.edu>,
        "histonet-bounces <@t> lists.utsouthwestern.edu"
        <histonet-bounces <@t> lists.utsouthwestern.edu>
Message-ID:
        <DACC27F195E63D44BDF7ADC5C52051AF1F02004AC2 <@t> DCPWVMBXC0VS1.mdanderson.edu>

Content-Type: text/plain; charset="us-ascii"

REMOVE FROM DATABASE

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim.Donadio <@t> bhcpns.org
Sent: Wednesday, July 07, 2010 8:37 AM
To: dcojita <@t> tampabay.rr.com
Cc: 'histonet'; histonet-bounces <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] used histology equipment

Hi Diane,
                    Because of your location I would recommend
Micro-optics of Florida. Mike Jones or Tom Christy. The number is
954-791-0082. They are great people to work with and I have found them
very reasonable on their prices.

Hope this helps!



Kim Donadio
Pathology Supervisor
Baptist Hospital
1000 W Moreno St.
Pensacola FL 32501
Phone (850) 469-7718
Fax (850) 434-4996



<dcojita <@t> tampabay.rr.com>
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
07/06/2010 05:29 PM

To
"'histonet'" <Histonet <@t> lists.utsouthwestern.edu>
cc

Subject
[Histonet] used histology equipment






Does anyone know of a reputable dealer for used equipment?

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet



-----------------------------------------
All electronic data transmissions originating from or sent to
Baptist Health Care Corporation (BHC) are subject to monitoring.
This message along with any attached data, are the confidential and
proprietary communications of BHC and are intended to be received
only by the individual or individuals to whom the message has been
addressed. If the reader of this message is not the intended
recipient, please take notice that any use, copying, printing,
forwarding or distribution of this message, in any form, is
strictly prohibited and may violate State or Federal Law. If you
have received this transmission in error, please delete or destroy
all copies of this message.  For questions, contact the BHC Privacy
Officer at (850) 434-4472.  Rev.10/07.
_______________________________________________
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------------------------------

Message: 20
Date: Wed, 07 Jul 2010 08:36:47 -0700
From: sgoebel <@t> xbiotech.com
Subject: [Histonet] Reprocess
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <20100707083647.9e2d9aa830e8449a2412eb1e4f2f067e.ed952ac3ac.wbe <@t> email04.secureserver.net>

Content-Type: text/plain; charset="utf-8"


   Hello  all!!  Hope everyone had a happy 4th!!  Question    grossed  in  some  fat  that I need to routine process.  I ha   this  in  the  past  with  extra fixation and no problem?  This tim   however  it didn't fix all the way through and I have oily unfixed fat
   in     section  we   longer  and rep   then  what?    older =)

   Thanks in advance!!

   Sarah Goebel, B.A., HT (
   Histotechnician

   XBiotech USA Inc.


   8201 East Riverside Dr. Bldg 4 Suite 100

   Austin, Texas  78744

   (512)386-5107


------------------------------

Message: 21
Date: Wed, 7 Jul 2010 11:47:39 -0400
From: Catherine Simonson <catherinesimonson <@t> gmail.com>
Subject: Re: [Histonet] Reprocess
To: sgoebel <@t> xbiotech.com
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
        <AANLkTinikHppsDePiCYWIKcKmGQ2YOtS3Ig0LgOUDJgS <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Easy option is to melt down, and run cassettes through the clean cycle on
your processors to remove all the paraffin.  then process as usual.  I've
done this before with decent results.

Good luck!
Catherine

On Wed, Jul 7, 2010 at 11:36 AM, <sgoebel <@t> xbiotech.com> wrote:

>
>   Hello  all!!  Hope everyone had a happy 4th!!  Question  today is...I
>   grossed  in  some  fat  that I need to routine process.  I ha ve done
>   this  in  the  past  with  extra fixation and no problem?  This tim e
>   however  it didn't fix all the way through and I have oily unfixed fat
>   in    the  middle  of  my  block.  I fixed for 48 hours, but guess my
>   section  we  re  too big and needed more.  I want to fix for a little
>   longer  and rep rocess the block.  I know I need to melt it down, but
>   then  what?   I  have done this before, it's just my brain is getting
>   older =)
>
>   Thanks in advance!!
>
>   Sarah Goebel, B.A., HT ( ASCP)
>
>   Histotechnician
>
>   XBiotech USA Inc.
>
>
>   8201 East Riverside Dr. Bldg 4 Suite 100
>
>   Austin, Texas  78744
>
>   (512)386-5107
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>


------------------------------

Message: 22
Date: Wed, 7 Jul 2010 10:53:18 -0500
From: Cheri Miller <cmiller <@t> physlab.com>
Subject: RE: [Histonet] Reprocess
To: "sgoebel <@t> xbiotech.com" <sgoebel <@t> xbiotech.com>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <E3C81A010935EA41B379AC765103F3BF32C00E235B <@t> olsrv12>
Content-Type: text/plain; charset="us-ascii"

Hi Sarah, I don't melt them down I just drop them in with my dirty molds and lids and run them through the cleaning cycle on my processor. The Xylene and Alcohol will melt the oily fat. When the clean cycle is complete I just drop them back into formalin until I process that evening. It works very well.


Cheryl A. Miller HT(ASAP)cm
Histology/Cytology Prep Supervisor
Physicians Laboratory Services
Omaha, NE. 402 731 4145 ext. 554

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of sgoebel <@t> xbiotech.com
Sent: Wednesday, July 07, 2010 10:37 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Reprocess


   Hello  all!!  Hope everyone had a happy 4th!!  Question =day is...I
   grossed  in  some  fat  that I need to routine process.  I ha= done
   this  in  the  past  with  extra fixation and no problem?  This tim=
   however  it didn't fix all the way through and I have oily unfixed fat
   in  =e  middle  of  my  block.  I fixed for 48 hours, but guess my
   section  we=  too big and needed more.  I want to fix for a little
   longer  and rep=cess the block.  I know I need to melt it down, but
   then  what? =ave done this before, it's just my brain is getting
   older =)

   Thanks in advance!!

   Sarah Goebel, B.A., HT (=CP)

   Histotechnician

   XBiotech USA Inc.


   8201 East Riverside Dr. Bldg 4 Suite 100

   Austin, Texas  78744

   (512)386-5107
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message.  If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information.  Any dissemination, distribution, or copying of this e-mail is strictly prohibited.  If you have received this message in error, please notify the sender immediately and delete this email from your system.



------------------------------

Message: 23
Date: Wed, 7 Jul 2010 11:01:37 -0500
From: Cheri Miller <cmiller <@t> physlab.com>
Subject: RE: [Histonet] Reprocess
To: Cheri Miller <cmiller <@t> physlab.com>, "sgoebel <@t> xbiotech.com"
        <sgoebel <@t> xbiotech.com>, "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <E3C81A010935EA41B379AC765103F3BF32C00E235D <@t> olsrv12>
Content-Type: text/plain; charset="us-ascii"

I take that back I do melt them, recap the cassettes and then drop them in to clean.

Cheryl A. Miller HT(ASAP)cm
Histology/Cytology Prep Supervisor
Physicians Laboratory Services
Omaha, NE. 402 731 4145 ext. 554

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Cheri Miller
Sent: Wednesday, July 07, 2010 10:53 AM
To: sgoebel <@t> xbiotech.com; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Reprocess

Hi Sarah, I don't melt them down I just drop them in with my dirty molds and lids and run them through the cleaning cycle on my processor. The Xylene and Alcohol will melt the oily fat. When the clean cycle is complete I just drop them back into formalin until I process that evening. It works very well.


Cheryl A. Miller HT(ASAP)cm
Histology/Cytology Prep Supervisor
Physicians Laboratory Services
Omaha, NE. 402 731 4145 ext. 554

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of sgoebel <@t> xbiotech.com
Sent: Wednesday, July 07, 2010 10:37 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Reprocess


   Hello  all!!  Hope everyone had a happy 4th!!  Question =day is...I
   grossed  in  some  fat  that I need to routine process.  I ha= done
   this  in  the  past  with  extra fixation and no problem?  This tim=
   however  it didn't fix all the way through and I have oily unfixed fat
   in  =e  middle  of  my  block.  I fixed for 48 hours, but guess my
   section  we=  too big and needed more.  I want to fix for a little
   longer  and rep=cess the block.  I know I need to melt it down, but
   then  what? =ave done this before, it's just my brain is getting
   older =)

   Thanks in advance!!

   Sarah Goebel, B.A., HT (=CP)

   Histotechnician

   XBiotech USA Inc.


   8201 East Riverside Dr. Bldg 4 Suite 100

   Austin, Texas  78744

   (512)386-5107
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message.  If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information.  Any dissemination, distribution, or copying of this e-mail is strictly prohibited.  If you have received this message in error, please notify the sender immediately and delete this email from your system.

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message.  If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information.  Any dissemination, distribution, or copying of this e-mail is strictly prohibited.  If you have received this message in error, please notify the sender immediately and delete this email from your system.



------------------------------

Message: 24
Date: Wed, 7 Jul 2010 11:24:51 -0500
From: "Mahoney,Janice A" <Janice.Mahoney <@t> alegent.org>
Subject: RE: [Histonet] Reprocess
To: "'sgoebel <@t> xbiotech.com'" <sgoebel <@t> xbiotech.com>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <8F0EE4144E8E2F4CA1F6B051A2E5BFEEA7C4A2F0 <@t> EXCHMBC2.ad.ah.local>
Content-Type: text/plain; charset="utf-8"

Cheri's process is a good one, we use it too.  Works every time.
Jan Mahoney
Omaha, NE

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of sgoebel <@t> xbiotech.com
Sent: Wednesday, July 07, 2010 10:37 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Reprocess


   Hello  all!!  Hope everyone had a happy 4th!!  Question =day is...I
   grossed  in  some  fat  that I need to routine process.  I ha= done
   this  in  the  past  with  extra fixation and no problem?  This tim=
   however  it didn't fix all the way through and I have oily unfixed fat
   in  =e  middle  of  my  block.  I fixed for 48 hours, but guess my
   section  we=  too big and needed more.  I want to fix for a little
   longer  and rep=cess the block.  I know I need to melt it down, but
   then  what? =ave done this before, it's just my brain is getting
   older =)

   Thanks in advance!!

   Sarah Goebel, B.A., HT (=CP)

   Histotechnician

   XBiotech USA Inc.


   8201 East Riverside Dr. Bldg 4 Suite 100

   Austin, Texas  78744

   (512)386-5107
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet


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