[Histonet] cell culture staining
Ian Birchall
IBirchall <@t> groupwise.swin.edu.au
Tue Oct 7 18:54:43 CDT 2003
Hi, after fixation with NBF I have performed H+E staining on the cells
still adherent to the culture plate. Following fixation, wash with
water, cover with haematoxylin (Harris) 5min, wash, blue (if necessary)
wash, cover with eosin (3 min), wash and coverslip (on the culture
plate) using aqueous mountant (Gurr, aquahere). You can also do immuno
staining for cell surface markers on cultured cells without removing
them from the plate. Regards, Ian Birchall, Melbourne.
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Today's Topics:
1. RE: Cell Culture Staining (C.M. van der Loos)
2. Re: Purchasing machines Grossing Work
Station,Microtom etc. (Fred Underwood)
3. Re: Purchasing machines Grossing Work
Station,Microtom etc. (Fred Underwood)
4. Nerve Tissue (LaTricia Faison)
5. Re: COX 1&2 (DDittus787 <@t> aol.com)
6. Hard Tissue Committee Meeting (Vicki Kalscheur)
7. Histotech needed (MGomez <@t> ameripath.com)
8. For the Guiness Book Of Records (George Cole)
--__--__--
Message: 1
Date: Tue, 07 Oct 2003 13:35:10 +0200
From: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>
To: histonet <@t> lists.utsouthwestern.edu
Cc: KJohnson <@t> med.miami.edu
Subject: [Histonet] RE: Cell Culture Staining
Hi Kevin,
There is a method described by Harold Kerstens to embed your cells into
paraffin blocks. Spin the cells down in an Eppendorf tube, remove the
supernatant and mix the cells gently with 1 ml warm (65C) 2% agarose
solution. Centrifuge again to concentrate the cells. Embed in paraffin
as usally.
See paper by: Kerstens et al. JHC 48:709-718, 2000.
For the next time try to get some collagen sponges from some cell
culturing company. Spin the (unfixed!) cells down and wash 3x with PBS
(remove proteins). Discard supernatant and resuspend cells in a small
volume. Insert sponge and allow the cells to adhere to the collagen (15
min, RT). Fix sponge in buffered formalin and embed as usual.
Good luck!
Chris van der Loos
Dept. of Cardiovascular Pathology
Academical Medical Center
Amsterdam - The Netherlands
>-----Original Message-----
>From: Johnson, Kevin [mailto:KJohnson <@t> med.miami.edu]
>Sent: Friday, October 03, 2003 3:29 PM
>To: 'histonet <@t> lists.utsouthwestern.edu'
>Subject: [Histonet] Cell Culture Staining
>
>Greetings, all.
>
>A researcher just brought me (at Miller Time on a Friday!) two
culture
>flasks of terminally differentiated Sertoli cells and requested H&E
>staining. Well.
>Conventional H&E obviously is out, since polystyrene is not compatible
>with xylene. Further restrictions: the cells cannot be scraped off,
>they cannot be replated on glass coverslips, and for reasons of her
>own, they cannot go back into the incubator pending an answer. They
>now have been fixed in situ with 10% neutral buffered formalin.
>
>Does anyone have a protocol for staining these puppies? If not H&E
>per se, then is there another staining method that might satisfy her?
>
>Thanks in advance,
>
>Kevin Johnson
>University of Miami
>Diabetes Research Institute
--__--__--
Message: 2
Date: Tue, 07 Oct 2003 08:42:04 -0400
From: "Fred Underwood" <funderwood <@t> mcohio.org>
To: <histonet <@t> lists.utsouthwestern.edu>,<histo <@t> skm.org.pk>
Subject: Re: [Histonet] Purchasing machines Grossing Work
Station,Microtom etc.
I would highly recommend the tissue tek DRS 2000 automated stainer and
to complement that the SCA coverslipper. Bothe are sold by Sakura
Finetek. I'm not sure of their web address but call 1.800.725.8723 to
get more product info.
Fred
>>> "Histology" <histo <@t> skm.org.pk> 10/01/03 11:23AM >>>
Our lab is in the process of purchasing our automated stainer for
routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center
and processor (300 cassettes). I would really appreciate comments from
anyone who really likes, or dislikes the machines that they are using.
Our plan is to get an automatic coverslipper as well.
Thanks in advance!!!
Muhammad Tahseen
Histology Supervisor
Deptt. Pathology
Shaukat Khanum Cancer Hospital
And Research Center Lahore. Pakistan
Ph. +92 42 5180725-36 Ext 2369,
Fax. +92 42 5180723
e-mail. Histology <@t> skm.org.pk
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--__--__--
Message: 3
Date: Tue, 07 Oct 2003 08:42:04 -0400
From: "Fred Underwood" <funderwood <@t> mcohio.org>
To: <histonet <@t> lists.utsouthwestern.edu>,<histo <@t> skm.org.pk>
Subject: Re: [Histonet] Purchasing machines Grossing Work
Station,Microtom etc.
I would highly recommend the tissue tek DRS 2000 automated stainer and
to complement that the SCA coverslipper. Bothe are sold by Sakura
Finetek. I'm not sure of their web address but call 1.800.725.8723 to
get more product info.
Fred
>>> "Histology" <histo <@t> skm.org.pk> 10/01/03 11:23AM >>>
Our lab is in the process of purchasing our automated stainer for
routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center
and processor (300 cassettes). I would really appreciate comments from
anyone who really likes, or dislikes the machines that they are using.
Our plan is to get an automatic coverslipper as well.
Thanks in advance!!!
Muhammad Tahseen
Histology Supervisor
Deptt. Pathology
Shaukat Khanum Cancer Hospital
And Research Center Lahore. Pakistan
Ph. +92 42 5180725-36 Ext 2369,
Fax. +92 42 5180723
e-mail. Histology <@t> skm.org.pk
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
--__--__--
Message: 4
Date: Tue, 07 Oct 2003 09:47:48 -0400
From: "LaTricia Faison" <lfaison <@t> mail.mcg.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Subject: [Histonet] Nerve Tissue
Does anybody know of a way to unthaw nerve that has been snap frozen to
a =
cork without doing damage to the nerve cells? It's hard to cross
section =
and remove the nerve from the cork when it is attached to it by snap =
freezing. Thanks in advance for any advice.
--__--__--
Message: 5
Date: Tue, 07 Oct 2003 10:25:11 -0400
From: DDittus787 <@t> aol.com
To: Luis.Chiriboga <@t> med.nyu.edu, dbpiontek <@t> hsc.wvu.edu,
histonet <@t> pathology.swmed.edu
Subject: Re: [Histonet] COX 1&2
you can try zymed as well, recently used both and they worked well.
1-800-874-4494
dana
--__--__--
Message: 6
From: "Vicki Kalscheur" <kalschev <@t> svm.vetmed.wisc.edu>
To: "Histonet Discussion" <histonet <@t> pathology.swmed.edu>
Date: Tue, 7 Oct 2003 11:13:19 -0500
Subject: [Histonet] Hard Tissue Committee Meeting
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Dear members and interested individuals,
Please join us on Saturday, October 18th, 2003 from 5:30 - 6:30 pm
=
in Louisville, KY as we gather at NSH's 29th Annual =
Symposium/Convention. The room number will be posted at the Hard
Tissue =
Display Table. I look forward to seeing many of you!=20
Sincerely,=20
Vicki Kalscheur, Chairperson
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<DIV><FONT face=3DArial size=3D2>Dear members and interested=20
individuals,</FONT></DIV>
<DIV><FONT face=3DArial size=3D2></FONT> </DIV>
<DIV><FONT face=3DArial size=3D2> Please join us on
=
Saturday,=20
October 18th, 2003 from 5:30 - 6:30 pm in Louisville, KY as we gather
at =
NSH's=20
29th Annual Symposium/Convention. The room number will be posted
=
at the=20
Hard Tissue Display Table. I look forward to seeing many of you! =
</FONT></DIV>
<DIV><FONT face=3DArial size=3D2></FONT> </DIV>
<DIV><FONT face=3DArial size=3D2>Sincerely, </FONT></DIV>
<DIV><FONT face=3DArial size=3D2> Vicki
Kalscheur,=20
Chairperson</FONT></DIV></BODY></HTML>
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--__--__--
Message: 7
From: MGomez <@t> ameripath.com
To: Histonet <@t> lists.utsouthwestern.edu
Date: Tue, 7 Oct 2003 12:11:54 -0400
Subject: [Histonet] Histotech needed
Good morning Histonetters:
We are looking for a certified HT/HTL to work at our Histology
Laboratory
located in Sandy, Utah. If you know anyone interested please write or
call
at 801-256-0040 x205/x244 for immediate consideration. You may ask
for
Milton or Mike.
We're located 20 minutes from Salt Lake City and 40 minutes from Park
City.
Thank you very much,
Milton
--__--__--
Message: 8
From: "George Cole" <georgecole <@t> ev1.net>
To: <histonet <@t> lists.utsouthwestern.edu>
Date: Tue, 7 Oct 2003 09:11:01 -0700
Subject: [Histonet] For the Guiness Book Of Records
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We have a place in the Guiness Book Of Records----Egad!! Who would
ever
think there would be so many histotechs named
Markus F. Meyenhofer.
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<p class=3DMsoNormal><font size=3D2 face=3DArial><span =
style=3D'font-size:10.0pt;
font-family:Arial'>We have a place in the <span =
class=3DSpellE>Guiness</span>
Book <span class=3DGramE>Of</span> Records----Egad!! Who would ever =
think there
would be so many <span class=3DSpellE>histotechs</span> =
named<o:p></o:p></span></font></p>
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style=3D'font-size:10.0pt;
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=
F. <span
class=3DSpellE>Meyenhofer</span>. <span =
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style=3D'mso-spacerun:yes'> </span><o:p></o:p></span></font></p>
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