Paula
You need to sit down with them and review the slides together, you can't solve
a problem without seeing the problem. They need to explain and show you what
they consider dirty.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303)
I would contact your local department of public health or your states governing
agency for hazardous/biological waste - for example in Colorado our hazardous
waste is controlled by the Colorado Department of Health. You should be able
to register a complaint through them.
On a personal note
Fred
We have a block trimmer here in the lab it's not the newcomer supply its from
Shandon, we like it. I personally do not use it but the rest of the
technicians do. I'm a bit old school I have a very old steak knife that I
have been using since 1996 so it's nice and dull and works
I might get some individuals upset but we really need to watch what we do in
the histology lab and how that effects the precision of our process. When you
breath on the block you are basically generating a thicker section, it's not a
good practice to keep. Trust me I used to do this in the
I'm going to give my two cents here. I think you need to look closely at how
you handle these samples. First of all leeps are not small samples therefore
just because the leep has been placed in fixative upon removal and then sits in
fixative until its processed. Unless the sample is grossed
Bob
I believe that both the B/R an CBG recyclers are fractional distillation units,
and to be honest I'm not even certain why I choose the CBG over B/R instrument
when I purchased. Prior to purchasing the CBG I was using the Suncycle
Technologies system, these less expensive systems are more
We have been recycling for years now and have not had any adverse effects. We
have a VIP 3 and a VIP 6. We do use fresh xylene for the cleaning cycle and we
only recycle our 95 and absolute alcohols along with both xylene and propar.
We use a fractional distillation unit from CBG. We have a
Bernice
There are lots of different markers for fibroblasts via IHC - If you are
looking for a good source Acris Antibodies has quite a few fibroblast markers
Vimentin,
Smooth Muscle Actin
FSP-1 (fibroblast specific protein)
Pro Collagen I
CD34
Prolyl-4-Hydroxylase beta
HSP-47
I have a two
You can also thaw and refreeze we have done that before.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com
Ship to Address:
Premier
Jessica
We have not tried Ram-11 in sheep it's a macrophage antibody against rabbit, in
our hands it did not cross react with human, cyno or rat. We use MAC387 to
stain macrophages in sheep. It's not entirely specific to macrophages but it
is what we have used in the past.
Liz
Elizabeth A.
Blanca
If your samples are fixed and processed properly the sections should cut
without wrinkles. We process mouse pancreas on a 30 minute cycle - 30 minutes
per station starting at 50% alcohol. We trim and soak blocks on ice and we
normally do not have issue with wrinkles.
Liz
Elizabeth
Paula
We have ours hooked up to tap water.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com
Ship to Address:
Premier Laboratory, LLC
1567
Jessica
There are a couple of things that we will try here when we have problems with
tissue adherence
1. We utilize a particular brand of plus slides - trubond 380 - these have
worked very well in our hands, we purchase them from Newcomer Supply
2. Post fix in 10% NBF for 10 minutes after
If you are not that concerned over the alcian blue portion of the movats you
can perform an elastic trichrome - that's what we do here. Mordant like you
would in Bouins, and rather than stain with weigerts hemastoxylin, stain with
the elastic portion of the VVG, differentiate like you would
We used the stain to stain nissl substance in mouse lumbar spine.
Purchased the dye from sigma - below is from the SDS Sheet
Product name : Gallocyanine
Product Number : 124508
Brand : Sigma-Aldrich
CAS-No. : 1562-85-2
The procedure in Bancroft-Gamble uses Chrom Alum - which is chromium
We have run this stain before, purchased the reagent from Sigma, it's pretty
easy to do there is a procedure in Bancroft and Gamble.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
We use freezer boxes and then record what is in the freezer boxes on a
worksheet that is kept in an excel file. The excel file has all of the boxes
in it.
Here is an example of one of the pages
BOX 6 - Full
name
vendor
catalog #
lot #
date alqt
ul/aliquot
notes
AGE
Biorbyt
Ronnie
We have these protocols in place for porcine tissue. We use the roche kit for
tunel 11684809910 and for CC3 we use the cell signaling antibody 9664.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060
Atoska
I'm pretty sure that the Sakura Tissue Tek Prisma has the capability to hold 2
x 3 slides with some adaptors to the slide baskets, I have a fax from 2009
that list the parts necessary. The regular 20 slide holders can be changed to
hold 5 - 2 x 3 slides.
Liz
Elizabeth A. Chlipala,
Are you using a good plus slide? We have started using these Tru-Bond slides
when we have tissue adherence problems, they seem to work really well. How are
you handling the slides once you section?
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO
Lauren
I have always used a very dull steak knife (sharp ones are too sharp and will
cut the cassette) mine is probably over 20 years old (now I'm dating myself).
I'm old school so I still use that but the rest of the techs will use the para
trimmer. The para trimmer is more ergonomically
James
We use recycled xylene in our H stainer and have never experienced any fading
of the eosin. When you recycle your clearing agent do you check for purity and
contamination, you have to also take into consideration that xylene substitutes
may behave differently than xylene. We have an
Linda
We have in our reagent policy that any chemical that does not come with an
expiration date - we assign it a 5 year from receipt expiration date.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
Cathy
Before doing that I would check the sequence homology of the immunogen between
human and rhesus. I would do this for all sources of the antibody, if the
company does not want to provide you with the sequence then technical support
should be able to blast the sequence for you and get
Gareth
I would think it would have more to do with OSHA than CDC - this could involve
several modules.
We have modules that cover the following - this is not all that we train on.
If you are a small business OSHA can help out. We have utilized their services
with a voluntary compliance
There are many companies that sell frozen tissue both normal and diseased.
Frozen IHC does not normally require retrieval most FFPE IHC does. I don't
think it would be good practice to do what you are suggesting.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box
Brett
Dako's rabbit anti human CD3 cross reacts with mouse, or where you looking for
a CD3 that only detects mouse?
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 881-0763 cell
(303) 682-9060 fax
and its attachments
without reading them or saving them to any media storage device.
-Original Message-
From: Elizabeth Chlipala via Histonet
[mailto:histonet@lists.utsouthwestern.edu]
Sent: Tuesday, May 31, 2016 9:50 AM
To: Patti Nelson - PNP Lab Consultant <nelsonr...@verizon.
Patti
We have the Sakura Prisma with the Glas coverslipper and we have ours vented
out.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com
Ship
The NSH as a part of its awards and scholarships program has the Newcomer
Helping Hand Scholarship.
If you are considering doing something like what Adelle or Ray did and are a
NSH member for two years you can apply for this scholarship. The scholarship
is for $2000.00 so it is quite
Nancy
I cannot comment on the plastic embedding but this also happens with paraffin
embedded samples. We trim off one of the edges on all mouse and rat eye
samples and that eliminates that problem with paraffin embedded samples. I can
send pictures of a rat eye that has been trimmed.
Liz
Gary
I think that might have to do with the fan motor inside the unit, it might need
to be replaced. We had ours replaced a few years ago. Do you service/PM the
cryostat yearly?
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303)
We have used the filtration systems in the past - the one thing you need to
keep in mind is that the percentage of alcohol you put in is the percentage you
get out. You need to test with a hydrometer and account for temperature what
you get out.So you only want to put in your absolute and
Teri
We have a spot on all of our IHC SOP's that will state if the antibody has been
tested on decalcified samples. We do not routinely add decalcified tissue to
our routine IHC protocol development runs, so for example if the target is a
soft tissue target we will develop on a variety of
Carl
We have tried multiple procedures, there are procedures that use a combination
of 2% osmium and 5% postassium dichromate, ones that call for shorter periods
of time in Osmium and then periodic acid rinse post osmium - this is referenced
in Freida's book - Histotechnology a
We use osmium post fixation to look at fat all of the time in mouse liver,
nerve and muscle samples. It works well, sample size needs to be thin, samples
are friable and can crack easily. We use a specific procedure for this it
includes potassium dichromate I think, I'm at home but on Monday
Judi
Through the years we have used a wide variety of software applications, we have
been performing image analysis since 2006, we started whole slide scanning in
2007. We have used various modules from Zeiss, Image Pro, Aperio's Toolbox,
Definiens and free shareware such at Image J. There
Bernice
We are not a clinical lab, we are a GLP compliant lab and we have a procedure
that addresses this and everything else about reagent preparation. I will put
the procedure and forms that we use on the NSH BLOCK for anyone who is
interested.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Eva
We have worked with various antibodies in order to detect human cells in a
rodent background. I know that this antibody has been referenced in the
literature and I also have known of other individuals who have had success with
it, but in our hands we have never been able to make it work
FYI – See below and while I have your attention there are lots of other Awards
and Scholarships that NSH offers its members. So nominate yourself or one of
your colleagues. It easy to do, all the information is at the link below.
http://www.nsh.org/content/nsh-awards-and-scholarships
Liz
Lin
The link to nominate one of your students for one of the NSH Student
Scholarship is below. Deadline for nomination is March 1, 2016. We have tried
to make the process easier this year for the program directors/education
coordinators the application process consists of two fillable forms
Hello Everyone
I have decided to post this since I have received some e-mails back regarding
the awards process. I have listed below my thoughts on Letters of
Recommendation.
Tips to Writing an Excellent Letter of Recommendation
The Awards Committee has to sift through multiple nominations
Lesley
We have a folder for each antibody, this folder contains all of the information
on that antibody. It includes all of our protocol development information, the
slides from protocol development in the flat plastic holders, spec sheets,
internet searches, publications and references. All
Mohamed
For larger samples we have ranges from 1.5 hours to 6 hours a station.
Anytime we have larger samples we always utilize three absolute alcohols and
three xylenes, for your samples I would try 1.5 to 2 hours per station to start.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier
I would second Terri's comments on the Sakura Glas coverslipper. I do have to
stress that yearly PM's are very important and that there is daily maintenance
that needs to be done in order for the instrument to function properly, we also
only use 24x50's.
Liz
Elizabeth A. Chlipala, BS,
Hello Histonetters
It's that time of the year again. Nominations for NSH Awards and Scholarships
is open.
Histotechs do great work every day. What better way to honor those leaders in
the field than by nominating them for an NSH award? Check out all of the
James
We use 10% Formic Acid to decal mouse femur and tibia - The rear legs are
removed at the hip with a pair of rongeurs the skin is removed to the ankle,
we allow the knee joints to fix for 48 hours, they need to be in a normal
degree of flexion, meaning you want them in a container that
Marcus
You can stain with H after DAB if you think it would help your how you
analyze your slides. DAB with the addition of a special stain has been
published and for us depending upon the project that we are working on we may
choose to use a different counterstain after an IHC that has DAB
That's correct if you do not quench endogenous peroxidase activity completely
you would be able to visualize the RBC's but maybe in instances like this it
might be better to have the rbc's stain a different color than the DAB they
would be easier to recognize, especially if they are
I have an operator/user manual that I can pdf for you but I do not have the
service manual.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
l...@premierlab.com
www.premierlab.com
Laura
They may not be processed enough. We process brain samples for both mouse and
rat 2 mm thick sections (cut on a brain matrix) on a 45 minute cycle. If they
are processing half of the mouse brain (these samples can be 4 -5 microns
thick). I don't think the cycle is long enough, that is
Jorge
I can comment on the cresyl etch violet - we actually purchased cresyl violet
acetate from MP Biomedicals. This particular chemical has not been certified
by the biological stain commission, if you are looking for a dye that has been
certified then I would go to Sigma-Aldrich - they
I'm not sure that is the case in the grand scheme of things, it will depend
upon the target that you need to stain via immunofluorescence. Technically we
perform IF staining on frozen sections primarily because the antigen does not
survive formalin fixation. Many people utilize IF techniques
Teri
Excellent point and to add to that most IF techniques that are employed on FFPE
are not direct they are indirect and consist of an unlabeled primary and then a
secondary that is conjugated with an alexa fluor or something similar at least
that is how we approach IF on FFPE tissues here.
We do both, we only have criteria for the temp but we do record both in all
rooms in the laboratory. We use traceable wall thermometers/humidity units and
replace upon expiration. We have a few SOP's that govern this, we are a GLP
compliant lab.
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Karen
There was a similar question on temp charts the other day on the histonet. We
record our temps daily on a form. Our temperature range is determined by the
equipment in the room, all operating manuals have ranges listed for temp, we
list all major equipment in a room and what the
I think you may a have a serious problem if "so much tissue washes off" if
that is happening you have problems with tissue adherence. A properly
processed and cut section should not wash off the slide, or even a portion of
it should not wash off. That would mean that you did not provide to
Tim
First of all my comment was not meant to criticize your post and even if you
may not think so I was trying to help. I stand by what I said, my comment was
to address what I thought was the amount of tissue loss your lab experiences
and if I took your comment too literally I apologize but
Tiana
We have never experienced any issue like that here we will use the HIER
solutions in the PT links for up to three times within a period of 2 weeks.
We have had our PT link units validated and they are calibrated yearly. Do you
review the Target Retrieval Run Reports, we print and keep
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