[Histonet] RE: purple blue haze
lharris <@t> samhealth.org
lharris <@t> samhealth.org
Fri Feb 10 16:01:31 CST 2006
We had a purple blue haze on our H&E's when we used cold tap water run through the stainer. We had to use hot water and the haze went away. I've tried to return to using cold water and we get the haze every time.
Lori A. Harris, HT (ASCP)
Histology Section Leader
GSRMC - Pathology
3600 NW Samaritan Drive
Corvallis, OR 97330
1-541-768-6078
lharris <@t> samhealth.org
-----Original Message-----
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Sent: Wednesday, December 21, 2005 8:06 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 25, Issue 26
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Today's Topics:
1. online journals (Malcolm McCallum)
2. Purple blue haze (Heckford, Karen - SMMC-SF)
3. Re: online journals (John A. Kiernan)
4. Re:Sections coming off! (Shannon Bryce)
5. re: MMA sections coming off! (Tracey Couse)
6. Re 17. fluorescence immunohistochemistry (Donna Harclerode)
7. Re: Purple blue haze (Andrea Grantham)
8. RE: Purple blue haze (Laurie Colbert)
9. Re: anti-human VEGF antibody (pruegg <@t> ihctech.net)
10. Re: 17. fluorescence immunohistochemistry (John Kiernan)
11. Re: non-specific staining IHC (Malam Jacqueline)
12. MMA sections falling off (Nancy W. Troiano)
13. Modified Davidson's Rat Eyes and Testes Fixation Times
(Sharon E Willman)
14. Embedding Centers (Roberta Horner)
15. Glass coverslippers (Angela Bitting)
16. RE: Glass coverslippers (Bartlett, Jeanine)
17. Re: Embedding Centers (Rene J Buesa)
18. San Antonio Histology connection (Jackie M O'Connor)
19. RE: Embedding Centers (Anne Van Binsbergen)
20. RE: Glass coverslippers (Anne Van Binsbergen)
21. Re: MMA sections falling off (Gayle Callis)
22. Re: Embedding Centers (Pamela Marcum)
23. decal rapid (Sharon Allen)
24. FW: artifact (Santana, Diane)
25. antibody search (Steven Coakley)
26. Mouse liver tissue processing (Luis Chiriboga)
27. RE: Mouse liver tissue processing (HSRL)
----------------------------------------------------------------------
Message: 1
Date: Tue, 20 Dec 2005 12:11:01 -0600
From: "Malcolm McCallum" <Malcolm.McCallum <@t> tamut.edu>
Subject: [Histonet] online journals
To: <parc <@t> listserv.uga.edu>, <ecolog-l <@t> listserv.umd.edu>,
<tws-l <@t> list.wildlife.org>, <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<EFF133CED606184A8FC9A6FBBF46714E013C91BF <@t> STAN.tamut.local>
Content-Type: text/plain; charset="iso-8859-1"
Hi
Which of the below three options do you think is best regarding a journal with both print and online versions????
1) Journal is open access, the publisher will charge page charges to authors.
2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access.
3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee.
In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article.
Thanks for the feedback, its actually very important!
Malcolm L. McCallum
Assistant Professor
Department of Biological Sciences
Texas A&M University Texarkana
2600 Robison Rd.
Texarkana, TX 75501
O: 1-903-233-3134
H: 1-903-791-3843
Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html
------------------------------
Message: 2
Date: Tue, 20 Dec 2005 11:17:21 -0700
From: "Heckford, Karen - SMMC-SF" <Karen.Heckford <@t> CHW.edu>
Subject: [Histonet] Purple blue haze
To: "Histonet (E-mail)" <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<E9A86CC7AD6D904D91CABCC05392933E031A1ED5 <@t> sfmh-msg-001.CHW.EDU.>
Content-Type: text/plain; charset="iso-8859-1"
Dear Histonetters,
Has anyone had any problems with a purple- blue haze on their H&E using
Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I
am still getting the haze. I have changed nothing, it just appeared a
couple of weeks ago. Any help would be greatly appreciated.
Happy Holidays,
Karen Heckford HT (ASCP) CE
Lead Histology Technician
St. Mary's Medical Center
Histology Department
450 Stanyan St.
San Francisco, Ca. 94117
1-415-668-1000 ext. 6167
email: kheckfor <@t> chw.edu
------------------------------
Message: 3
Date: Tue, 20 Dec 2005 13:42:34 -0500
From: "John A. Kiernan" <jkiernan <@t> uwo.ca>
Subject: Re: [Histonet] online journals
To: Malcolm McCallum <Malcolm.McCallum <@t> tamut.edu>
Cc: histonet <@t> lists.utsouthwestern.edu, tws-l <@t> list.wildlife.org,
ecolog-l <@t> listserv.umd.edu, parc <@t> listserv.uga.edu
Message-ID: <43A8509A.9B855C2A <@t> uwo.ca>
Content-Type: text/plain; charset=us-ascii
You raise some interesting points. Clearly you're
polling a wide range of scientists; please send
out a summary when you've analysed the results.
Here's my two-centsworth.
Paying to read the full text doesn't apply if your
institution subscribes to the journal (or more
usually to a large batch from the publisher).
Since most people who want the full text are in
institutions with libraries (or with employers
such as drug companies that will pay for
individual articles), avoiding page charges is the
way to go. Why should the researcher and author
pay the publisher? It should be the other way
round. Prestigious journals commonly do charge
authors, but anyone publishing in such a journal
is sure to have a big enough research grant to
cover the fees, which are small compared with
salaries and items for the lab.
Some journals published by learned societies do
not charge authors anything, even for coloured
pictures if these are needed to make a point.
Three examples that come to mind are the Journal
of Anatomy, the Journal of Histochemistry and
Cytochemistry, and Biotechnic & Histochemistry.
I'm sure there are many others. All three that I
mentioned come free with membership in the
society. The cost works out at about $5 per issue
(print, and access online).
I've not heard of the option of the author paying
and the reader not paying. It's hard to see that
working; even non-profit societies cannot run
their journals at a loss. Libraries have to pay
much more than individual members.
--
-------------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
kiernan[AT]uwo.ca
http://publish.uwo.ca/~jkiernan/
http://instruct.uwo.ca/anatomy/530/index.htm
_______________________________
Malcolm McCallum wrote:
>
> Hi
> Which of the below three options do you think is best regarding a journal with both print and online versions????
>
> 1) Journal is open access, the publisher will charge page charges to authors.
> 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access.
> 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee.
>
> In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article.
>
> Thanks for the feedback, its actually very important!
>
> Malcolm L. McCallum
> Assistant Professor
> Department of Biological Sciences
> Texas A&M University Texarkana
> 2600 Robison Rd.
> Texarkana, TX 75501
> O: 1-903-233-3134
> H: 1-903-791-3843
> Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 4
Date: Tue, 20 Dec 2005 13:01:17 -0600
From: "Shannon Bryce" <Shannon.Bryce <@t> UTSouthwestern.edu>
Subject: [Histonet] Re:Sections coming off!
To: "Joanne Mauger" <mauger <@t> email.chop.edu>,
<Histonet <@t> lists.utsouthwestern.edu>,<plott <@t> uab.edu>
Message-ID: <43A8009D0200009900000BC0 <@t> swnw124.swmed.edu>
Content-Type: text/plain; charset=US-ASCII
We get our slides which are the + coated slides from Fisher. Our MMA
sections are also falling off here lately so I went to another
department and borrowed a box of theirs that they bought through VWR and
have had no problems so I would guess it's a bad lot.....anyone have lot
numbers?
Thanks,
Shannon Bryce
UT Southwestern Medical Center
>>> "Joanne Mauger" <mauger <@t> email.chop.edu> 12/20/05 8:39 AM >>>
Hi Patricia,
Are you using slides from Erie? We buy ours through Fisher, and have
been having trouble keeping sections on + slides as well. It is as if
they have no charge at all. We think the lot may be bad. The company
claims to have no complaints.
Anyone else having similar problem?
Jo Mauger
>>> "Patricia F Lott" <plott <@t> uab.edu> 12/19/05 3:21 PM >>>
Help! We are losing our thin plastic sections! We have cut PMMA
plastic sections at 5 microns and put them on "+" slides, and the
sections fall off either during de-plasticizing or during staining!
We
have tried: Haupt's adhesive, longer drying times, "+" slides, plus
slides and Haupt's together, etc....no luck! Any suggestions would be
welcome!
_______________________________________________
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: 5
Date: Tue, 20 Dec 2005 15:15:42 -0500
From: Tracey Couse <tracey.couse <@t> ibb.gatech.edu>
Subject: re: [Histonet] MMA sections coming off!
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <43A8666E.2030005 <@t> ibb.gatech.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed
Patricia,
Try manually coating plain glass slides with poly-l-lysine,
chrome/gelatin, or glycerin/gelatin. We use all of these for our thin
plastic sections. I also suggest setioning a micron or two thinner (cut
at 3 or 4um). A thinner section will reduce the surface area to volume
ratio and may promote better section adherance. You may need to dry the
sldies using a slide press and dry longer or with higher temperatures.
Also, try searching the Histonet archives for alternative slide coating
suggestions. Good luck!
--
Tracey Couse
Laboratory Coordinator
Georgia Tech/Emory Center for the
Engineering of Living Tissues
Georgia Institute of Technology
IBB, Room 1123
Office: 404.385.2611
Lab: 404-385-6735
Fax: 404.894.229
Date: Mon, 19 Dec 2005 14:21:07 -0600
From: "Patricia F Lott" <plott <@t> uab.edu>
Subject: [Histonet] MMA sections coming off!
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<C4BBA8D04B6FAD47B10C5AFF4BBB93FD026F32EE <@t> UABEXMB2.ad.uab.edu>
Content-Type: text/plain; charset="us-ascii"
Help! We are losing our thin plastic sections! We have cut PMMA
plastic sections at 5 microns and put them on "+" slides, and the
sections fall off either during de-plasticizing or during staining! We
have tried: Haupt's adhesive, longer drying times, "+" slides, plus
slides and Haupt's together, etc....no luck! Any suggestions would be
welcome!
1
------------------------------
Message: 6
Date: Tue, 20 Dec 2005 13:06:17 -0800
From: "Donna Harclerode" <dharclerode <@t> cytoritx.com>
Subject: [Histonet] Re 17. fluorescence immunohistochemistry
To: <histonet <@t> lists.utsouthwestern.edu>,
<Emily.Wiesner <@t> medecine.unige.ch>
Cc: Adam Johnson <ajohnson <@t> cytoritx.com>
Message-ID:
<3DE0F644E093DF4BAE80C254176696A505B869 <@t> mp-mailserver.macropore.com>
Content-Type: text/plain; charset="US-ASCII"
Message: 17
Date: Mon, 19 Dec 2005 22:08:48 +0100
From: Emily.Wiesner <@t> medecine.unige.ch
Subject: [Histonet] fluorescence immunohistochemistry
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <1135026528.43a72160c086f <@t> webmail.medecine.unige.ch>
Content-Type: text/plain; charset=ISO-8859-1
Hi All,
I am new to fluorescence immunohistochemistry and I was wondering if
anyone can let me know the simple steps involved in performing this
technique. I know the dilutions required for the primary and secondary
antibodies, I just need to known what solutions are used for washes and
the steps in between!
Thanks in advance,
Emily
Dear Emily
I do very simple IHC using fluorescent secondaries from Jackson
Immunoresearch and purified primary abs. Depending on what you are
doing, there can be more steps added.
I usually am staining sections on a slide or cells on chamber slides or
in a plate.
For the unfixed frozen sections on slides I normally would fix 5 min in
75% acetone 25% ethanol and then right into PBS- do not allow the fixed
slides to dry. The only change would be if the antibody prefers
formalin fixation to acetone/ alcohol (thanks Gayle- I love this
fixative for almost everything).
I load them into a Sequenza(tm) rack (Thermo Electron, formerly Shandon)
or you could put them in a humid chamber of some sort, wiping excess
buffer before adding antibody or blocking solutions.
OPTIONAL I have recently started using Image-iT(tm) FX signal enhancer,
I36933, $80 Invitrogen for auto fluorescence (not sure it helps yet, but
sure does not hurt) 30 minutes. I would not use this on routinely- my
sections have infracted areas that have been a problem in the past)
After blocking, I rinse 1 time in the Sequenza rack (3 x 5 minutes if
you are doing them by hand)
Add 100ul of the antibody diluted in Dako antibody diluent S0809-
usually 1:100- 1:1k (dilutions are antibody dependant, but I have found
in fluorescence too high a concentration is not as big a problem as with
HRP formats)
If you are using a humid chamber be sure to cover all the tissue.
I have incubated slides at Room temperature for 1-2 hours, but now am
staining overnight in the fridge (not so good in clinical situations)
After primary incubation, I rinse 1 time in the Sequenza rack (3 x 5
minutes if you are doing them by hand)
Next is Jackson secondary at a 1:100 dilution (I use Cy3 conjugates for
single color and FITC and RITC for dual color. All my secondaries are
minimal cross (mouse to rat), but if all your tissue is human that is
not necessary. 30 minutes
I add DAPI to my secondaries to give a blue nuclear stain so I can tell
what I am looking at and localize the stain.
Rinse again 3 times in PBS and coverslip with your choice of aqueous
media. For media that never gets hard, I prefer Immunomount (Thermo
Electron, formerly Shandon) or Aqua Mount (Lerner from VWR) same thing
different label or the Prolonge Mounting media P36930 for $101 that will
harden. I do not like the hard mount media from Vector- I got many air
bubbles forming day after the sldes were coverslipped.
For cells on plates or chamber slides I usually fix with 10% NBF or 4%
PFA (same concentration of fixative, 4%PFA will contain no methanol) for
5-30 minutes. CD makers can be very hard to stain with NBF but with a
short fix, I have been pretty luck so far. I rinse 3 x 10 minutes with
PBS
I have not used the blocking reagents on cells as I have not seen any
autofluorescence in our cardiac myocytes, neurons or stem cells.
The rest of the procedure is the same except in a 6 well plate you need
a lot of antibody to cover the plate (about 600u). I try to use a
rocker if it is available to help to be sure of good coverage.
I do not usually cover my slides or plates, but if your room is very
bright or your signal very week, it would be a good idea to protect the
secondaries from light.
I like to use isotype controls (negative controls) or rabbit IgGs for
much of my work, but in a pinch I use just the antibody diluent to be
sure the stain I see, is a result of the primary antibody and not the
secondary binding to some part of the cell. In cell assays I always want
to stain a known positive and a known negative cell if at all possible
in addition to the isotype controls.
If you are making your own diluent (to me Dako diluent is great and I
have used it successfully for over 10 years in many applications)
I use 0.3% Triton, 5 % Normal serum from the species that your secondary
is made in - (all my secondaries are made in donkey so I would use
donkey serum) in PBS. I have used donkey secondaries when possible
because when I tested them years ago, they were a bit cleaner on the CD
markers I was using.
Dako diluent must have a similar make up, but it lasts a year in the
fridge and it permeablizes intact cells very nicely.
One big thing when purchasing fluorophores, make sure the filters on
your scope can see the secondaries you are purchasing. I found out at
my current job after staining with Cy3 and RITC that the reason I could
not get any stain was they had a Texas red filter not the Cy3, RITC
filter. Ideally if you are doing multicolor you want narrow band
filters so you can separate the colors, with broad pass filters the
colors blur together and it is hard to separate the results.
Good Luck!
Donna
Donna Harclerode, HT, (ASCP), HTL, QIHC
Scientist / Immunohistochemistry
Cytori Therapeutics
3020 Callan Rd.
San Diego, CA 92121
858-458-0900 ext 322
dharclerode <@t> cytoritx.com
------------------------------
Message: 7
Date: Tue, 20 Dec 2005 14:19:47 -0700
From: Andrea Grantham <algranth <@t> u.arizona.edu>
Subject: Re: [Histonet] Purple blue haze
To: "Histonet (E-mail)" <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<4.3.2.7.2.20051220140523.00ce4520 <@t> algranth.inbox.email.arizona.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
Karen,
Yes I have recently noticed the "haze" of the H&E stained slides. I have
been using this hematoxylin for years and this is the first time that I
have had a problem with it. Like you I have filtered it but it hasn't
seemed to help.
I also have been using their Type 6 paraffin and in the last few months
have noticed that it blows apart on the waterbath. I have lowered the temp.
but it didn't help -> 35-39 degrees C. A different lot number that they
sent to me was only marginally better. A couple of weeks ago one of my
clients brought a project consisting of blocks that had been processed and
embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The
blocks from the first 4 years cut fine and laid out on the waterbath fine
but when I got to recently embedded blocks they started the rapid expanding
and blowing apart when they were placed on the waterbath. A histotech in CA
told me they have been having the same problem in their lab.
Hmmmmm.....could they have changed the formula?
Maybe this is so with the Hematoxylin as well.
I'm trying different paraffins looking for one comparable to the Type 6
only more stable on the waterbath. I really don't want to do this with
Hematoxylin because my clients love my staining but I hate ugly slides.
Andi
At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote:
>Dear Histonetters,
>Has anyone had any problems with a purple- blue haze on their H&E using
>Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I
>am still getting the haze. I have changed nothing, it just appeared a
>couple of weeks ago. Any help would be greatly appreciated.
>
>Happy Holidays,
>
>
>Karen Heckford HT (ASCP) CE
>Lead Histology Technician
>St. Mary's Medical Center
>Histology Department
>450 Stanyan St.
>San Francisco, Ca. 94117
>1-415-668-1000 ext. 6167
>email: kheckfor <@t> chw.edu
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
.....................................................................
: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy :
: Sr. Research Specialist University of Arizona :
: (office: AHSC 4212) P.O. Box 245044 :
: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA :
: (FAX: 520-626-2097) (email: algranth <@t> u.arizona.edu) :
:...................................................................:
http://www.cba.arizona.edu/histology-lab.html
------------------------------
Message: 8
Date: Tue, 20 Dec 2005 15:12:17 -0800
From: "Laurie Colbert" <laurie.colbert <@t> huntingtonhospital.com>
Subject: RE: [Histonet] Purple blue haze
To: "Andrea Grantham" <algranth <@t> u.arizona.edu>, "Histonet (E-mail)"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<0BE6ADFAE4E7E04496BF21ABD346628006307C76 <@t> EXCHANGE1.huntingtonhospital.com>
Content-Type: text/plain; charset="iso-8859-1"
Your discussion rang a bell with me, and I've been meaning to call Richard Allan. We use their Type 9 paraffin, and for the last 3 or 4 months it also has been "blowing apart on the waterbath." We lowered the temp on the waterbaths, and this seems to have helped. Our housekeeping personnel have even noticed a difference in the paraffin (scraping it off the floor). They say it is stickier lately. I think something has been changed.
Laurie Colbert
Huntington Hospital
Pasadena, CA
-----Original Message-----
From: Andrea Grantham [mailto:algranth <@t> u.arizona.edu]
Sent: Tuesday, December 20, 2005 1:20 PM
To: Histonet (E-mail)
Subject: Re: [Histonet] Purple blue haze
Karen,
Yes I have recently noticed the "haze" of the H&E stained slides. I have
been using this hematoxylin for years and this is the first time that I
have had a problem with it. Like you I have filtered it but it hasn't
seemed to help.
I also have been using their Type 6 paraffin and in the last few months
have noticed that it blows apart on the waterbath. I have lowered the temp.
but it didn't help -> 35-39 degrees C. A different lot number that they
sent to me was only marginally better. A couple of weeks ago one of my
clients brought a project consisting of blocks that had been processed and
embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The
blocks from the first 4 years cut fine and laid out on the waterbath fine
but when I got to recently embedded blocks they started the rapid expanding
and blowing apart when they were placed on the waterbath. A histotech in CA
told me they have been having the same problem in their lab.
Hmmmmm.....could they have changed the formula?
Maybe this is so with the Hematoxylin as well.
I'm trying different paraffins looking for one comparable to the Type 6
only more stable on the waterbath. I really don't want to do this with
Hematoxylin because my clients love my staining but I hate ugly slides.
Andi
At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote:
>Dear Histonetters,
>Has anyone had any problems with a purple- blue haze on their H&E using
>Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I
>am still getting the haze. I have changed nothing, it just appeared a
>couple of weeks ago. Any help would be greatly appreciated.
>
>Happy Holidays,
>
>
>Karen Heckford HT (ASCP) CE
>Lead Histology Technician
>St. Mary's Medical Center
>Histology Department
>450 Stanyan St.
>San Francisco, Ca. 94117
>1-415-668-1000 ext. 6167
>email: kheckfor <@t> chw.edu
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
.....................................................................
: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy :
: Sr. Research Specialist University of Arizona :
: (office: AHSC 4212) P.O. Box 245044 :
: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA :
: (FAX: 520-626-2097) (email: algranth <@t> u.arizona.edu) :
:...................................................................:
http://www.cba.arizona.edu/histology-lab.html
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 9
Date: Tue, 20 Dec 2005 17:56:17 -0800 (PST)
From: pruegg <@t> ihctech.net
Subject: Re: [Histonet] anti-human VEGF antibody
To: "Katri Tuomala" <katri <@t> cogeco.ca>
Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <43434.207.200.116.10.1135130177.squirrel <@t> 207.200.116.10>
Content-Type: text/plain;charset=iso-8859-1
I am using mouse anti-vegf from Santa Cruz with good results on human and
baboon ffpe tissues, it is not easy, but I do HIER in a water bath in high
ph buffer at 85dc for 3 hrs., followed by overnight incubation of the
primary 1:200, detected with mouse labeled polymer (I use Envision from
DAKO).
Patsy
> Andrea,
> I cannot say this is the best anti-VEGF antibody, but I am doing a
> research
> project on FFPE human brain tissue and using placenta as a control with
> following antibody:
> monoclonal mouse anti human VEGF, clone VG1 from ID Labs (www.idlabs.com),
> dilution 1:100, HIER in citrate buffer pH 6.0. Detection system is from
> Zymed, Histostain Plus Kit..
> Glioblastoma stains strongly with this antibody. I have not used this with
> frozen sections.
>
> Katri
>
> Katri Tuomala
> Hamilton, Ontario, Canada
> ----- Original Message -----
> From: "Andrea T. Hooper" <anh2006 <@t> med.cornell.edu>
> To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
> Sent: Monday, December 19, 2005 8:37 PM
> Subject: [Histonet] anti-human VEGF antibody
>
>
>> What's the current consensus on the best antibody to use for anti-human
>> VEGF immunostaining on human FFPE sections (frozens as well if
>> possible).
>>
>> Thanks so much!!
>> Andrea
>> --
>>
>> _______________________________________________
>> 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: Wed, 21 Dec 2005 02:10:48 -0500
From: John Kiernan <jkiernan <@t> uwo.ca>
Subject: Re: [Histonet] 17. fluorescence immunohistochemistry
To: Donna Harclerode <dharclerode <@t> cytoritx.com>
Cc: histonet <@t> lists.utsouthwestern.edu, Adam Johnson
<ajohnson <@t> cytoritx.com>, Emily.Wiesner <@t> medecine.unige.ch
Message-ID: <43A8FFF8.D988E7C6 <@t> uwo.ca>
Content-Type: text/plain; charset=us-ascii
When I was a student in Birmingham, England
(that's the Birmingham with the silent h), the
immunology and serology lecturer told us:
"water is unphysiological and beer is expensive,
so we use saline." He must have been a good
lecturer because I can still recall that
apophthegm 40 years on when afar and asunder!
John Kiernan
London, Ontario.
-------------------------------
> Hi All,
> I am new to fluorescence immunohistochemistry and I was wondering if
> anyone can let me know the simple steps involved in performing this
> technique. I know the dilutions required for the primary and secondary
> antibodies, I just need to known what solutions are used for washes and
> the steps in between!
> Thanks in advance,
------------------------------
Message: 11
Date: Wed, 21 Dec 2005 12:00:58 -0000
From: Malam Jacqueline <Jacqueline.Malam <@t> rli.mbht.nhs.uk>
Subject: [Histonet] Re: non-specific staining IHC
To: "Histonet submissions (histonet <@t> lists.utsouthwestern.edu)"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <F2D584187C727B4CBFE008435881E8970D074D <@t> rlixch>
Content-Type: text/plain
We use the Benchmark XT but make our own prep kits up for neg controls (poly
and mono) and are not experiencing any more non-sp staining than we normally
do which is minimal.
Jacqui
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------------------------------
Message: 12
Date: Wed, 21 Dec 2005 07:20:53 -0500
From: "Nancy W. Troiano" <nancy.troiano <@t> yale.edu>
Subject: [Histonet] MMA sections falling off
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <5.2.1.1.2.20051221071248.00c2c038 <@t> email.med.yale.edu>
Content-Type: text/plain; format=flowed; charset=us-ascii
We use Chrome-alum gel coated slides and prepare them as
follows: dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled water
(solution A) while gently heating solution. Dissolve 4 g Chromium
Potassium Sulfate (Fisher #C337-500) in distilled water (solution
B). Prepare gelatin solution for slide dip: Mix 500 ml Solution a with
19.25 ml solution B. Heat to 70-75 Deg. C, dip slides 2 -4 minutes in this
solution. Allow to dry vertically, then put in hot oven overnight (60 deg.
C). To adhere sections to slides we place the 5 micron MMA sections on
slide, spread with 70% ethanol, cover with a piece of plastic and stack
slides then clamp with an office clamp. Put slides in 37deg C oven for two
nights or for one night in 60 deg C oven (don't do the hot oven if you are
subsequently staining for enzymes). For immuno we mount our 5 micron
plastic sections on hybridization slides from Scientific Device Laboratory,
Cat. #063 and place in hot oven 60 deg C overnite. Hope this helps!
------------------------------
Message: 13
Date: Wed, 21 Dec 2005 07:18:32 -0600
From: Sharon E Willman <sharon.willman <@t> bms.com>
Subject: [Histonet] Modified Davidson's Rat Eyes and Testes Fixation
Times
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <43A95628.7060609 <@t> bms.com>
Content-Type: text/plain; format=flowed; charset=ISO-8859-1
Hi,
I was needing input on how long you fix rat eyes and testes in Modified
Davidson's. Any suggestions would be most appreciated.
Thanks in advance,
Sharon Willman
------------------------------
Message: 14
Date: Wed, 21 Dec 2005 08:25:26 -0500
From: "Roberta Horner" <rjr6 <@t> psu.edu>
Subject: [Histonet] Embedding Centers
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<A6B73F11A6EC22409C4271EF2BB48A544CD34F <@t> ADLMAIL.padlspsu.psu.edu>
Content-Type: text/plain; charset="US-ASCII"
My embedding center just died. Last time our maintenance looked at it
they couldn't get the parts (its about 17 years old). I need to get
information quick on which ones you like and are good. So far I have
quotes from Surgipath and on the Leica but I don't know anything about
them. Any comments please?
Roberta
------------------------------
Message: 15
Date: Wed, 21 Dec 2005 08:49:01 -0500
From: "Angela Bitting" <akbitting <@t> geisinger.edu>
Subject: [Histonet] Glass coverslippers
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <s3a91710.023 <@t> GHSGWIANW4.GEISINGER.EDU>
Content-Type: text/plain; charset=US-ASCII
Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks.
IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you.
------------------------------
Message: 16
Date: Wed, 21 Dec 2005 08:56:05 -0500
From: "Bartlett, Jeanine" <jqb7 <@t> cdc.gov>
Subject: RE: [Histonet] Glass coverslippers
To: "Angela Bitting" <akbitting <@t> geisinger.edu>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<CB857F6460D42E4AAEA195054A25406C0C7775BB <@t> m-ncid-2.ncid.cdc.gov>
Content-Type: text/plain; charset="us-ascii"
We recently demo'd the new one from Leica and it was great!
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Angela
Bitting
Sent: Wednesday, December 21, 2005 8:49 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Glass coverslippers
Can anyone recommend a automated glass coverslipper? I've used Hacker in
the past. Any that are faster, better? Thanks.
IMPORTANT WARNING: The information in this message (and the documents
attached to it, if any) is confidential and may be legally privileged.
It is intended solely for the addressee. Access to this message by
anyone else is unauthorized. If you are not the intended recipient, any
disclosure, copying, distribution or any action taken, or omitted to be
taken, in reliance on it is prohibited and may be unlawful. If you have
received this message in error, please delete all electronic copies of
this message (and the documents attached to it, if any), destroy any
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 17
Date: Wed, 21 Dec 2005 06:12:16 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Embedding Centers
To: Roberta Horner <rjr6 <@t> psu.edu>, histonet <@t> lists.utsouthwestern.edu
Message-ID: <20051221141216.20482.qmail <@t> web61215.mail.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Hi Roberta:
I would suggest you to call Sakura-Finetek. They have a new embedding center that
has all the advantages of the old Tissue-Tek, along with the Sakura reliability.
Ask for a "demo" instrument (they usually accomodate that type of request).
René J.
Roberta Horner <rjr6 <@t> psu.edu> wrote:
My embedding center just died. Last time our maintenance looked at it
they couldn't get the parts (its about 17 years old). I need to get
information quick on which ones you like and are good. So far I have
quotes from Surgipath and on the Leica but I don't know anything about
them. Any comments please?
Roberta
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------------------------------
Message: 18
Date: Wed, 21 Dec 2005 08:25:29 -0600
From: "Jackie M O'Connor" <Jackie.O'Connor <@t> abbott.com>
Subject: [Histonet] San Antonio Histology connection
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <OF426B975B.B4C72078-ON862570DE.004F1B7E <@t> abbott.com>
Content-Type: text/plain; charset="us-ascii"
Could someone in the San Antonio area contact me directly?
Thanks.
Jackie O'
------------------------------
Message: 19
Date: Wed, 21 Dec 2005 18:20:37 +0400
From: "Anne Van Binsbergen" <vanann702 <@t> skmc.gov.ae>
Subject: RE: [Histonet] Embedding Centers
To: "Roberta Horner" <rjr6 <@t> psu.edu>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<7E070A7B959A9F42BE732545E5CF621094ACBA <@t> SKMCEMAIL.skmc.gov.ae>
Content-Type: text/plain; charset="us-ascii"
Im a fan of the Sakura embedding system - it's ergonomically designed
and VERY user friendly
Can be 'left handed' or 'right handed'
Cassettes left/right
Moulds left/right
Cold plate left/right
Forceps left/right
Nothing more annoying (being a lefty) to be forced to work like a
non-lefty!!
Everything works independently of everything else - ie - micro
adjustments can be made to temp in almost any area to your requirements
Nice broad anti burn protection to rest hands/fingers while embedding
And if you have a VIP5 the basket fits snugly into the holding area -
left or right as you prefer of course!!
Like I said - I am THE fan of Sakura
And I just KNOW that some of you are smiling out there - you know who
you are!!!
Annieinarabia
-----Original Message-----
From: Roberta Horner [mailto:rjr6 <@t> psu.edu]
Sent: Wednesday, December 21, 2005 5:25 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Embedding Centers
My embedding center just died. Last time our maintenance looked at it
they couldn't get the parts (its about 17 years old). I need to get
information quick on which ones you like and are good. So far I have
quotes from Surgipath and on the Leica but I don't know anything about
them. Any comments please?
Roberta
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 20
Date: Wed, 21 Dec 2005 18:24:12 +0400
From: "Anne Van Binsbergen" <vanann702 <@t> skmc.gov.ae>
Subject: RE: [Histonet] Glass coverslippers
To: "Bartlett, Jeanine" <jqb7 <@t> cdc.gov>, "Angela Bitting"
<akbitting <@t> geisinger.edu>, <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<7E070A7B959A9F42BE732545E5CF6210948C5F <@t> SKMCEMAIL.skmc.gov.ae>
Content-Type: text/plain; charset="us-ascii"
Try the Sakura
Fast, efficient, small 'footprint' - will fit into a small space
Had a Leica once - was so very patient - 2 exchanges (new machines)
later - still hated it
Annieinarabia
-----Original Message-----
From: Bartlett, Jeanine [mailto:jqb7 <@t> cdc.gov]
Sent: Wednesday, December 21, 2005 5:56 PM
To: Angela Bitting; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Glass coverslippers
We recently demo'd the new one from Leica and it was great!
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Angela
Bitting
Sent: Wednesday, December 21, 2005 8:49 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Glass coverslippers
Can anyone recommend a automated glass coverslipper? I've used Hacker in
the past. Any that are faster, better? Thanks.
IMPORTANT WARNING: The information in this message (and the documents
attached to it, if any) is confidential and may be legally privileged.
It is intended solely for the addressee. Access to this message by
anyone else is unauthorized. If you are not the intended recipient, any
disclosure, copying, distribution or any action taken, or omitted to be
taken, in reliance on it is prohibited and may be unlawful. If you have
received this message in error, please delete all electronic copies of
this message (and the documents attached to it, if any), destroy any
hard copies you may have created and notify me immediately by replying
to this email. Thank you.
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
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------------------------------
Message: 21
Date: Wed, 21 Dec 2005 08:14:11 -0700
From: Gayle Callis <gcallis <@t> montana.edu>
Subject: [Histonet] Re: MMA sections falling off
To: "Nancy W. Troiano" <nancy.troiano <@t> yale.edu>,
Histonet <@t> lists.utsouthwestern.edu
Message-ID:
<6.0.0.22.1.20051221081017.01b39c08 <@t> gemini.msu.montana.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
To increase the holding power, the gelatin can be 275 bloom, a larger
gelatin molecule made from pig collagen. This subbing solution suggested
by Nancy is tried and true, also works for the nasty decalcified bone
sections, generally larger ones. However, if the subbing solution causes
too much blue background staining from hematoxylin, one can dip the subbed
slides in NBF a couple of times, rinse well, dry and store in cool, dry
place. The NBF crosslinks the gelatin a bit and reduces blue background.
At 05:20 AM 12/21/2005, you wrote:
>We use Chrome-alum gel coated slides and prepare them as
>follows: dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled
>water (solution A) while gently heating solution. Dissolve 4 g Chromium
>Potassium Sulfate (Fisher #C337-500) in distilled water (solution
>B). Prepare gelatin solution for slide dip: Mix 500 ml Solution a with
>19.25 ml solution B. Heat to 70-75 Deg. C, dip slides 2 -4 minutes in
>this solution. Allow to dry vertically, then put in hot oven overnight
>(60 deg. C). To adhere sections to slides we place the 5 micron MMA
>sections on slide, spread with 70% ethanol, cover with a piece of plastic
>and stack slides then clamp with an office clamp. Put slides in 37deg C
>oven for two nights or for one night in 60 deg C oven (don't do the hot
>oven if you are subsequently staining for enzymes). For immuno we mount
>our 5 micron plastic sections on hybridization slides from Scientific
>Device Laboratory, Cat. #063 and place in hot oven 60 deg C
>overnite. Hope this helps!
>
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)
------------------------------
Message: 22
Date: Wed, 21 Dec 2005 10:14:58 -0500
From: Pamela Marcum <pmarcum <@t> vet.upenn.edu>
Subject: Re: [Histonet] Embedding Centers
To: "Roberta Horner" <rjr6 <@t> psu.edu>,
<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <6.1.1.1.2.20051221101125.01997318 <@t> mail.vet.upenn.edu>
Content-Type: text/plain; charset="us-ascii"; format=flowed
At 08:25 AM 12/21/2005, Roberta Horner wrote:
>My embedding center just died. Last time our maintenance looked at it
>they couldn't get the parts (its about 17 years old). I need to get
>information quick on which ones you like and are good. So far I have
>quotes from Surgipath and on the Leica but I don't know anything about
>them. Any comments please?
>Roberta
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Hi,
I will be totally different as i am buying the Thermo Shandon Embedding
Center. I had it in for demo and it had all the features the others have
for timing and pre-sets as well as some independent temperature ranges I
likes. Also the cold plate is seperate so I can have it on either side or
in another area if I want for special projects. Good Luck, I think they
all work well it just depends for me on features and price and Shandon also
had the right price.
Best Regards,
Pamela A Marcum
Manager, Histology Special Procedures
University of Pennsylvania
School of Veterinary Medicine
R.S. Reynolds Jr. CORL
New Bolton Center
382 West Street Road
Kennett Square, PA 19348
Phone - 610-925-6278
Fax - 610-925-8120
E-mail - pmarcum <@t> vet.upenn.edu
------------------------------
Message: 23
Date: Wed, 21 Dec 2005 09:17:14 -0600
From: Sharon Allen <SAllen <@t> exchange.hsc.mb.ca>
Subject: [Histonet] decal rapid
To: "Histonet (E-mail)" <histonet <@t> pathology.swmed.edu>
Message-ID:
<36FE435D6D2F1D489174B22362A961B66B830B <@t> hscxntmx0006.hsc.mb.ca>
Content-Type: text/plain; charset="iso-8859-1"
Hi, Does anyone know about or what supplier "Decal Rapid" by "Pathtech" can
be purchased from?
We had used it a few years ago but have no idea where we got it from. I
tried searching the internet & came up with Pathtech in Australia but would
like a supplier a little closer.
Thanks in advance
Sharon
sallen <@t> hsc.mb.ca
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
------------------------------
Message: 24
Date: Wed, 21 Dec 2005 10:27:06 -0500
From: "Santana, Diane" <dsantana <@t> pmaonline.com>
Subject: [Histonet] FW: artifact
To: "'Histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB04D7E282 <@t> MAILPMA>
Content-Type: text/plain; charset="iso-8859-1"
> -----Original Message-----
> From: Santana, Diane
> Sent: Tuesday, December 20, 2005 10:20 AM
> To: 'histonet <@t> lists.utsouthwestern.edu'
> Subject: artifact
>
> I am having a problem with artifact on some of my GI bx's. Actually a
> paper published by Anatech "The Innovator", summer2004 shows this same
> artifact on the front page. They believe it is a fixation problem, and or
> retrieval by the endo Drs. My pathologist rules both these ideas out. I
> tend to agree with him, at lease on the fixation. Our bx's go into the
> processor around 5 pm and the bx run starts around 2 am.
> It is a loss of nuclear detail. This artifact can be on a small section of
> a bx. Even if there is 5 bx's in one cassette maybe 1 will have this
> artifact where the others look great. I thought at first it was a clearing
> problem or nuclear bubbling I changed times in clear rite and oven. It did
> not eliminate the problem. My pathologist has seen this by other labs
> also, but very seldom. For us it could be 1 or 2 slides a day. Sometimes
> not at all. Weekend or weekdays. No consistently. The best way I can
> describe it is that it looks like a smudge.
> Our bx run is this;
>
> 10% NBF 30 min ea x2
> 80% Alcohol 10mins
> 95% alcohol 10 mins ea x2
> 100% alcohol 17 mins ea x3
> Clear rite 22 mins ea x2
> Paraffin 15 mins ea x3
>
> I would appreciate any feedback on this.
> Diane Santana
> Pentucket Medical Assoc.
> Haverhill, Mass.
------------------------------
Message: 25
Date: Wed, 21 Dec 2005 07:22:41 -0800 (PST)
From: Steven Coakley <sjchtascp <@t> yahoo.com>
Subject: [Histonet] antibody search
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID: <20051221152241.11885.qmail <@t> web90204.mail.scd.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Gooday everyone,
I'd like to get a few recommendations for anti-insulin and anti-glucagon antibodies for FFPE mouse tissue. If anyone has used a specific companies antibodies that work well please let me know. I'm still learning this research IHC stuff so I can use all the help I can "muster"
Steve
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------------------------------
Message: 26
Date: Wed, 21 Dec 2005 09:54:28 -0500
From: Luis Chiriboga <Luis.Chiriboga <@t> med.nyu.edu>
Subject: [Histonet] Mouse liver tissue processing
To: Histonet <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <KFEIIJOCLBABEKFDAEHPOEKCFFAA.Luis.Chiriboga <@t> med.nyu.edu>
Content-Type: text/plain; charset="iso-8859-1"
Hi everyone
We are having a real serious problem getting good sections from mouse
livers. The tissue curls and shreds out of the block as soon as it comes in
contact with the knife, end up with a section that has the exact outline of
the tissue but no tissue. The tissue consistency could best be described as
"dry?" Interestingly, it is only the normal livers that have this problem.
We have tried sectioning thick, thin, no soak, cold soak......but out of
ideas. Anyone have any suggestions? Unfortunately we have no control over
the tissue processing, but perhaps can convince to change.
Thanks Luis
Happy Holidays to all!!
------------------------------
Message: 27
Date: Wed, 21 Dec 2005 11:02:33 -0500
From: "HSRL" <histosci <@t> shentel.net>
Subject: RE: [Histonet] Mouse liver tissue processing
To: "'Luis Chiriboga'" <Luis.Chiriboga <@t> med.nyu.edu>,
<histonet <@t> pathology.swmed.edu>
Message-ID: <000c01c60647$f47a7ab0$0200a8c0 <@t> HSRLMAIN>
Content-Type: text/plain; charset="us-ascii"
Luis,
Have a tried keeping them on a WET ice for 45 minutes to an hour? If
you have tried it and it doesn't work, it is a processing problem that
you must rectify.
-Tom
Tom Galati
Laboratory Director
HSRL, Inc.- A GLP Compliant Contract Laboratory
5930 Main Street
Mount Jackson, Virginia 22842
540.477.4440
540.477.4448
tomgalati <@t> hsrl.org
www.hsrl.org
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Luis
Chiriboga
Sent: Wednesday, December 21, 2005 9:54 AM
To: Histonet
Subject: [Histonet] Mouse liver tissue processing
Hi everyone
We are having a real serious problem getting good sections from mouse
livers. The tissue curls and shreds out of the block as soon as it
comes in
contact with the knife, end up with a section that has the exact outline
of
the tissue but no tissue. The tissue consistency could best be described
as
"dry?" Interestingly, it is only the normal livers that have this
problem.
We have tried sectioning thick, thin, no soak, cold soak......but out of
ideas. Anyone have any suggestions? Unfortunately we have no control
over
the tissue processing, but perhaps can convince to change.
Thanks Luis
Happy Holidays to all!!
_______________________________________________
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
End of Histonet Digest, Vol 25, Issue 26
****************************************
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