[Histonet] Help with immunohistochemistry in human embryos

megan twomey megantwomey <@t> hotmail.com
Fri Nov 10 10:56:35 CST 2006


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       ______________________________________________________________

     From:  "Amy Porter" <portera <@t> msu.edu>
     To:  "Hernan Aldana"
     <haldana <@t> unimoron.edu.ar>,<histonet <@t> lists.utsouthwestern.edu>
     Subject:  Re:  [Histonet]  Help  with immunohistochemistry in human
     embryos
     Date:  Fri, 10 Nov 2006 10:24:11 -0500
     >You might want to try using a Heat Epitope Induced Retrieval
     >protocol.  Most of Santa Cruz's antibodies will work with heat
     >retrieval.
     >Amy S. Porter, HT (ASCP) QIHC
     >Investigative HistoPathology Laboratory - Supervisor
     >2201 Biomedical Physical Sciences Bldg.  Rm #2133
     >East Lansing, MI  48824-3320
     >Phone:  (517) 355-6475 ext 1480
     >Fax:  (517) 432-1368
     >Email:  portera <@t> msu.edu
     >Web:  www.humanpathology.msu.edu
     >----- Original Message ----- From: "Hernan Aldana"
     ><haldana <@t> unimoron.edu.ar>
     >To: <histonet <@t> lists.utsouthwestern.edu>
     >Sent: Thursday, November 09, 2006 10:45 AM
     >Subject:   [Histonet]  Help  with  immunohistochemistry  in  human
     embryos
     >
     >
     >Dear Histoneters
     >I  need some advices. I'm working with human embryos between 7 and
     14
     >weeks.
     >we used Sonic Hedgehog Shh (C-18) and sc-1195 antibodies of Santa
     >Cruz
     >Biotechnology in paraffin sections of embryos fixed in
     >paraformadehyde. I
     >don't have good results.
     >Could  anybody  help me with some advice, concerning to every step
     of
     >my
     >technique?
     >
     >Thanks in advance
     >
     >Dr. Hernán Javier Aldana Marcos
     >
     >Prof. Titular Reg. Histología, Embriología y Biología Celular
     >
     >
     >Facultad de Medicina
     >Universidad de Morón
     >Argentina
     >
     >email altenrativo: hernanjavier <@t> yahoo.com
     >http://www.ht.org.ar/educacion.htm
     >
     >
     >
     >
     >
     >-----Mensaje original-----
     >De: histonet-bounces <@t> lists.utsouthwestern.edu
     >[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] En nombre de
     >histonet-request <@t> lists.utsouthwestern.edu
     >Enviado el: Domingo, 01 de Octubre de 2006 02:04 p.m.
     >Para: histonet <@t> lists.utsouthwestern.edu
     >Asunto: Histonet Digest, Vol 35, Issue 1
     >
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     >When  replying,  please  edit  your  Subject  line  so  it is more
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     >
     >
     >Today's Topics:
     >
     >   1. Job Opening (Luck, Greg D.)
     >   2. RE: Problem of Double immunoflouresence staining ..
     >      (Melissa Gonzalez)
     >   3. The procedure that substitues xylene with mineral oil.
     >      (PhiHo Hoang)
     >   4. Re: The procedure that substitues xylene with mineral oil.
     >      (Rene J Buesa)
     >
     >
     >------------------------------------------------------------------
     ----
     >
     >Message: 1
     >Date: Sat, 30 Sep 2006 12:01:02 -0700
     >From: "Luck, Greg D." <LuckG <@t> empirehealth.org>
     >Subject: [Histonet] Job Opening
     >To: <histonet <@t> lists.utsouthwestern.edu>
     >Cc:      "Ringwood,     James  F."     <RingwoJ <@t> empirehealth.org>,
     "Wakabayashi,
     >Marlene T." <WakabaM <@t> empirehealth.org>, "Browning, Rachel M."
     ><BrowniR <@t> EmpireHealth.org>
     >Message-ID:
     ><6BB8BC4519AAB844B174FC739A679BBC50900C <@t> IRMEXCH01.irm.inhs.org>
     >Content-Type: text/plain; charset="us-ascii"
     >
     >Hello All,
     >
     >Deaconess  is  a  300 bed acute care community medical center.  We
     have
     >a
     >full   time,  M-F,  day  time  position  open.  Must  be  HT(ASCP)
     certified
     >or
     >eligible.  Duties include all routine histology functions in a
     >clinical
     >setting (i.e. embedding , cutting, special stains and frozen
     >sections).
     >IHC  experience  preferred.  Due  to a recent market adjustment by
     our
     >organization the salary range for this position has just increased
     >25-33%  depending on years of experience.  This is a golden
     >opportunity
     >for  someone  to  improve  on  their current situation or to begin
     anew.
     >Anyone new to this field is encouraged to apply as well, for the
     >door is
     >wide  open.  You  can  apply  on-line  by  going  to our corporate
     website
     >www.empirehealth.org.  The position is at Deaconess Medical Center
     >in
     >Spokane, WA www.deaconess-spokane.org.  Please contact me directly
     >for
     >more details.  Following is an additional web link to provide you
     >with
     >more info on Spokane and the greater Inland Northwest region
     >www.visitspokane.com. ("Near Perfect-Near Nature").  Thank you and
     >best
     >wishes, Greg
     >
     >Greg Luck, BS, HT(ASCP)
     >Anatomic Pathology Supervisor
     >Deaconess Medical Center
     >800 W. 5th Ave
     >Spokane, WA 99204
     >Phone 509.473.7077
     >Fax 509.473.7133
     >luckg <@t> empirehealth.org
     >www.deaconess-spokane.org
     >
     >
     >
     >------------------------------
     >
     >Message: 2
     >Date: Sun, 1 Oct 2006 08:40:44 -0700
     >From: "Melissa Gonzalez" <Melissa.Gonzalez <@t> cellgenesys.com>
     >Subject:   [Histonet]  RE:  Problem  of  Double  immunoflouresence
     staining
     >..
     >To: <histonet <@t> lists.utsouthwestern.edu>
     >Message-ID:
     ><A6B0506C2B000E45A7556B9A93C45547695866 <@t> hqsvr01mail.cgi.com>
     >Content-Type: text/plain; charset="us-ascii"
     >
     >Sohail,
     >First, Perhaps you should try incubating each primary with
     >respective
     >secondary  only,  to  determine how they look separately, and also
     make
     >sure  each  is  working  on its own. For example, you may not have
     your
     >aSMA
     >dilution  correct.  Also, do you have a positive control for TRITC
     to
     >make
     >sure you can detect any TRITC signal adequately? Once you have
     >appropriate signal from both, you can then make your cocktails.
     >Also, in
     >the future, I would recommend switching to Alexa Fluors from
     >Molecular
     >Probes,  they  really  are  great.  In your instance you would use
     AF488
     >(FITC) and AF 546 or 555 (TRITC). But that is just my 2 cents.
     >
     >Good luck,
     >Melissa
     >
     >
     >
     >Message: 4
     >Date: Fri, 29 Sep 2006 13:06:27 -0700 (PDT)
     >From: sohail ejaz <sohail_e <@t> yahoo.com>
     >Subject:  [Histonet]  Problem of Double immunoflouresence staining
     of
     >retinal vasculature
     >To: histonet <@t> lists.utsouthwestern.edu
     >Message-ID: <20060929200627.949.qmail <@t> web39506.mail.mud.yahoo.com>
     >Content-Type: text/plain; charset=iso-8859-1
     >
     >Hello everybody
     >
     >  I have a big problem of staining retina vasculature using a
     >cocktail
     >of two different antibodies. The problem is that i can only see
     >staining
     >with FITC but not with TRITC, hence i cant make a merger of both
     >FITC
     >and TRITC.
     >
     >  Cocktail  of  primary  antibodies  include (1) Monoclonal Rabbit
     Anti
     >human vWF (2) Monoclonal mouse Anti alpha SMA and cocktail of
     >secondary
     >antibodies include (1)  Anti rabbit FITC (2) Anti mouse TRITC
     >
     >  Please let me know your commentc to slove this issue.
     >
     >  Dr.Sohail
     >
     >
     >
     >
     >
     >------------------------------
     >
     >Message: 3
     >Date: Sun, 1 Oct 2006 11:47:56 -0400
     >From: "PhiHo Hoang" <phiho.hoang <@t> rogers.com>
     >Subject: [Histonet] The procedure that substitues xylene with
     >mineral
     >oil.
     >To: <histonet <@t> lists.utsouthwestern.edu>
     >Message-ID: <005b01c6e570$f725b820$f300a8c0 <@t> ttth>
     >Content-Type: text/plain; charset="iso-8859-1"
     >
     >
     >Greetings,
     >
     >I am a new subscriber to this list and I am reading
     >the list archive.
     >
     >I came across a posting from Reni J. responding to
     >a request from Sharon:
     >
     ><Quote>
     >
     >Sharon:
     >  I  am  sending  privately  a  procedure  I have that substitutes
     xylene
     >with
     >mineral oil. It is 100 times less toxic than xylene and 3 times
     >cheaper.
     >  Reni J.
     >
     >Sharon Allen <SAllen <@t> exchange.hsc.mb.ca> wrote:
     >
     >Does anyone use xylol substitutes, if so how good are they?
     >We do CJD testing & "the powers that be" don't like to pay for
     >getting rid
     >of contaminated xylol. I don't want to have to start testing
     >different
     >substitutes, so any help would be greatly appreciated.
     >Thanks
     >Sharon
     >sallen <@t> hsc.mb.ca
     >
     ></Quote>
     >
     >I am interested in this procedure.
     >
     >It is very much appreciated if I can get a copy
     >of that procedure.
     >
     >Best regards,
     >
     >PhiHo
     >
     >
     >
     >
     >
     >------------------------------
     >
     >Message: 4
     >Date: Sun, 1 Oct 2006 09:55:53 -0700 (PDT)
     >From: Rene J Buesa <rjbuesa <@t> yahoo.com>
     >Subject: Re: [Histonet] The procedure that substitues xylene with
     >mineral oil.
     >To: PhiHo Hoang <phiho.hoang <@t> rogers.com>,
     >histonet <@t> lists.utsouthwestern.edu
     >Message-ID: <20061001165553.32345.qmail <@t> web61219.mail.yahoo.com>
     >Content-Type: text/plain; charset=iso-8859-1
     >
     >PhiHo:
     >  I am attaching the procedure under separate cover.
     >  Reni J.
     >
     >PhiHo Hoang <phiho.hoang <@t> rogers.com> wrote:
     >
     >Greetings,
     >
     >I am a new subscriber to this list and I am reading
     >the list archive.
     >
     >I came across a posting from Reni J. responding to
     >a request from Sharon:
     >
     >
     >
     >Sharon:
     >I am sending privately a procedure I have that substitutes xylene
     >with
     >mineral oil. It is 100 times less toxic than xylene and 3 times
     >cheaper.
     >Reni J.
     >
     >Sharon Allen exchange.hsc.mb.ca> wrote:
     >
     >Does anyone use xylol substitutes, if so how good are they?
     >We do CJD testing & "the powers that be" don't like to pay for
     >getting rid
     >of contaminated xylol. I don't want to have to start testing
     >different
     >substitutes, so any help would be greatly appreciated.
     >Thanks
     >Sharon
     >sallen <@t> hsc.mb.ca
     >
     >
     >
     >I am interested in this procedure.
     >
     >It is very much appreciated if I can get a copy
     >of that procedure.
     >
     >Best regards,
     >
     >PhiHo
     >
     >
     >
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     >End of Histonet Digest, Vol 35, Issue 1
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