[Histonet] Help with immunohistochemistry in human embryos

Thu Nov 9 09:45:39 CST 2006

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
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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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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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