No, because the glycine acts by reducing the autofluorescene of the free
aldehydes (maybe Dr. Kiernan or another knowledgeable person in the chemistry
can tell us precisely how) rather than reducing the binding of other staining
components to the aldehydes.
-Original Message-
From: Bob
I tried earlier to send, for this thread, a wonderful paper by Nauta,
chronicaling the history his discovery of his tract tracing method, in which
serendipity and degradation of formalin played critical roles, not realizing
that the size of the attachment would prevent it from going through, so
If this works by binding free aldehyde groups that attach to antibodies/ or
fluorochromes, or biotinylated whatever. shouldn't it also work for DAB or
ABC immunolabeling and
reduce background labeling?
Bob
UCLA / VA Medical Center
On Fri, Dec 12, 2008 at 2:08 PM, Gayle Callis wrote:
> To reduce
Andrea,
We stain for murine dendritic cells very frequently. CD11c (HL3 clone) is
excellent from BD Biosciences using fresh snap frozen tissue with spleen as
the positive control. Fixation is with our beloved 25% ethanol/75% acetone
for 5 min at RT then going directly to buffer from fixativ
Dear Smit You wrote: I am a graduate student at University of Illinois,
chicago. I have been having tremendous problems with sectioning of paraffin
embedded rat maxilla with molars and I
searched the Histonet archives and found that people here have invaluable
experience, insight and suggestio
I am staining CA19-9 on pancreas and have a good amount of mucin trapping
the DAB. Is there a product that either blocks mucin or something else that
will limit the mucin effect?
Aprill Watanabe, B.S.
Research Associate
Integrated Cancer Genomics Division
Tissue Microarray Center (TMA)
Translatio
Is anyone staining for mouse dendritic cells either in paraffin or
frozen sections? If so, what markers/antibodies are you using?
Thanks, ANDREA
--
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To reduce aldehyde induced autofluorescence, you can use 100 - 300 mM glycine
in pH 7.4 buffer. TRIS buffer or even Dulbeccos PBS will work. You rehydrate
the section and then immerse into the glycine solution for 20 minutes, maybe
even longer. Glycine works by getting rid (binding?) of free
I believe that the "swiss cheese" holes are due to ice crystal formation
during freezing, at least that's the rationale we were always taught for
using additional fixation in sucrose-formalin after the initial fixation in
formalin, i.e. the sucrose would prevent ice crystal formation. Susan
--
Would anyone using the Biogenix Xmatrix be willing to speak with me
about your experience? I'd appreciate your feedback!
Thanks, j
Joyce Weems
Pathology Manager
Saint Joseph's Hospital
5665 Peachtree Dunwoody Rd NE
Atlanta, GA 30342
678-843-7376 - Phone
678-843-7831 - Fax
Confidenti
Dewaxing in one change of solvent, regardless of time in the solvent, is not a
good idea. After a rack of slides has passed through the first xylene, that
dish no longer contains xylene. It contains a solution of paraffin in xylene,
and with each additional rack, that paraffin solution becomes
Anyone doing IHC for CD123 and TCL-1 on formalin-fixed, paraffin-embedded
tissue?
Richard
Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596
(860) 545-0174 Fax
Just two things about crosslinking:
1- there is not such a thing as over crosslinkage, because when all the
reaction sites have reacted, there are no reaction sites left to "overreact";
and
2- formaldehyde fixation is a reversible reaction and the tissue will lose the
croslinkage even by placing
I used the MW oven to heat-boost the slutions but the actual staining was done
in a water bath. Why don't you try a water bath at 60ºC?
rené J.
--- On Fri, 12/12/08, TIMOTHY MALLOY wrote:
From: TIMOTHY MALLOY
Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS
To: "HISTONET ..." , "TIM MALLOY"
D
So I have a question about the cross-linking aspect of PFA...while I agree
I need it to keep my epitope in place, is there such a thing as
OVER-crosslinking (i.e., tissue spending TOO much time in formalin - weeks?
months?) that would make my epitope difficult to near-impossible to
retrieve?
Why not use the (30-9) clone? It is much easier to come by. I sue it on my
Benchmark, cat# 790-4286.
On Fri, Dec 12, 2008 at 1:45 PM, Sebree Linda A wrote:
> Good afternoon, 'netters,
>
> I am in desperate need of Ki67 antibody, clone 7B11 preferably, to use
> on Ventana instruments. I didn't re
Good afternoon, 'netters,
I am in desperate need of Ki67 antibody, clone 7B11 preferably, to use
on Ventana instruments. I didn't realize we were running low and our
usual vendor has no stock. I'd prefer a predilute but only if it is
known to work on our instruments. I don't have the time to se
i have had this argument with the researchers at the University f= or
30 years, somewhere back in the day they were told that commercially
mad= e formalin had methanol in it (it does but just a little and does
not hurt = anything in my experience) and that methanol would damage
I will be out of the office starting 12/12/2008 and will not return
until 12/15/2008.
I will respond to your message when I return. Any histology-related
requests or questions can be sent to: histol...@wlgore.com.
___
Histonet mailing l
Hi,
I am a graduate student at University of Illinois, chicago. I have been having
tremendous problems with sectioning of paraffin embedded rat maxilla with
molars and I
searched the Histonet archives and found that people here have invaluable
experience, insight and suggestions for various probl
Hi - Here we perform the silver step with a hot plate under the hood. As the
slides are going into the sodium metabisulfite step, we turn on a hot plate on
high. Then we mix the silver solution according to the procedure, put it in a
beaker on the hot plate, rinse our slides with microwaved hot
If you are interested in knowing the concentration of formaldehyde,
i.e., making sure you have 10% formalin, you can contact CBG Biotech at
800-941-9484. They sell and service one of the best solvent recyclers I
have ever used. When I was at Rockford Memorial Hospital, we used one
of their recycl
I will be out of the office starting 12/12/2008 and will not return until
12/15/2008.
If you require immediate assistance, please contact Michelle Broome at x
47477.
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http://lists.utsouthwester
Oh, I'd like to know that, too, please!
--On Friday, December 12, 2008 7:48 AM -0600 "Walters, Katherine S"
wrote:
This may be another silly question, but how does one test the
concentration of formaldehyde in solution?
Thanks,
Kathy
Notice: This UI Health Care e-mail (including attachm
http://www.astm.org/Standards/D2194.htm
ASTM D2194 - 02(2007)
ASTM D2194 - 02(2007) Standard Test Method for Concentration of Formaldehyde
Solutions
2008-12-12
tf
发件人： Walters, Katherine S
发送时间： 2008-12-12 21:49:26
收件人： ti...@foxmail.com; Tony Henwood; Pat Flannery;
histo...@list
In research lab situations particularly, one does not have the time or
technique for nailing down the ways of making each of the buffers,
reagents, and procedures work the "right" way or the most optimum way...a
lot of times it's students or postdocs just focused on getting their
project done a
Matt may have said no but we still found out!
Linda Blazek HT (ASCP)
Manager/Supervisor
GI Pathology of Dayton
7415 Brandt Pike
Huber Heights, OH 45424
Phone: (937) 293-4424 ext 7118
Email: lbla...@digestivespecialists.com
-Original Message-
From: histonet-boun...@lists.utsouthwestern.
Joe, I can hardly believe you could be that grouchy :). When I was
looking hard for a tech I called my buddy Matt Chase at Children's to
try and coax him over to my lab. When he said no, I asked if he would
let me chat with his techs and see how they were doing. Alas, he also
replied negatively
You hit the nail on the head "That's what we always use", fear of change
is a common human condition.
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pat Flannery
Sent: 11 December 2008 16:59
To: h
This may be another silly question, but how does one test the concentration of
formaldehyde in solution?
Thanks,
Kathy
Notice: This UI Health Care e-mail (including attachments) is covered by the
Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and
may be legally p
We have used Paraplast Plus for years. For both processing and
embedding.
Betsy Molinari HT(ASCP)
Texas Heart Institute
Cardiovascular Pathology
6770 Bertner Ave
MC 1-283
Houston, TX 77030
832-355-6524
832-355-6812 (fax)
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[
Not an immediate help, but there will be a NSH teleconference May 27, 2009
on embedding.
PARAFFIN EMBEDDING AND PROCESS IMPROVEMENT
Date: May 27, 2009
Time: 1:00PM EST
Presented by Joelle Weaver, HTL(ASCP), Blanchard Valley Hospital, Findlay,
Ohio; Riverside Methodist Hospital, Columbus, Ohio; His
We currently use a stain called grocotts silver methenimine solution for
staining pnuemocystis. We would like to use a hot plate for the procedure
instead of a unvented microwave. Does anybody have any formula's the would like
to share with the hot plate?
Timothy G. Malloy, HT ( ASCP ) A.A.S.
"I looked at the sections and the cell shrinkage (and prominent spaces
between cells and connective tissue) indicated that most of the
"fixation" seemed to have occured in the processing ethanols. I asked
him for some of the fixative he used, tested the formaldehyde
concentration and found it to be
Hi, Because wash out solution can be 0.9% saline or 0.01 M PBSnear to
physiological osmotic pressure...
However the perfusion solution (4% PFA in 0.1M PB) is more concentrated, in the
sense of osmotic pressureCan you comment on this?
You can note the shrinkage of brain after PFA perfusi
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