uot; to get correct red amyloid that is green (dichroic, not fluorescent)
>> with crossed polars?
>> *John Kiernan*
>>
>> https://www.schulich.uwo.ca/anatomy//people/faculty/emeriti/kiernan_john.html
>> = = =
>> --
>> *From:* Greg Dobbin v
method were they using "at another
> lab" to get correct red amyloid that is green (dichroic, not fluorescent)
> with crossed polars?
> *John Kiernan*
>
> https://www.schulich.uwo.ca/anatomy//people/faculty/emeriti/kiernan_john.html
> = = =
>
Hello experts,
*Some background:*
I know that Congo Red can bind nonspecifically to non-amyloid components
such as collagen and elastin under certain conditions (eg Carnoys fixative,
insufficient differentiation, insufficient alkalinity, etc). However,
everything I have been able to read on the top
Hi Kara,
Use multi-tissue block controls with normal tissues that are treated
exactly the same (pre-analytically speaking) as your routine surgical
specimens. Consult NordiQC.org for recommended normal control tissue for
each IHC marker. The most used multi-tissue control block in my lab has
pieces
Decant formalin off into formalin waste containers and the remaining tissue
go into a red bag (red signifying destined for incineration). As the O.R.
does with fresh tissues for disposal.
*Greg Dobbin*
1205 Pleasant Grove Rd
Route 220
York, PE C0A 1P0
Use a formic acid based decal solution and MAKE SURE the specimen does not
fix (formalin fixation) longer than 32hrs (false negatives result). Use 70%
ETOH to manage the fixation time (ie fix for 24 hours, decal for 4 hrs,
place in 70% ETOH until it gets put on the processor. Hold in 70% ETOH for
w
Hi Nancy,
You should have no problem doing so. If you want proof, just run a couple
of your IHC control sections through the H&E stainer as you would for a
patient specimen (so right through to coverslipping) and then start the
process of removing coverslip, hydration, destain in acid alcohol and t
We use the same. Our protocol is ER2 for 15 mins. We have 3 washes after
polymer.
Greg
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A 1P0
*If God made a drink better than whiskey he kept it for himself! *
___
Histonet mailing list
Histone
Since images cannot be attached, if any of you feel you have some
experience that I may find helpful...please email me directly and I will
attach the image to my reply.
Thanks again,
*Greg Dobbin*
Chief Technologist
Anatomic Pathology
Queen Elizabeth Hospital
Charlottetown, PE, Canada
Hello experts,
Every now and then we will get stained slides that look like the attached
image. We run one processing schedule (about 11 hrs) for everything so yes,
our smaller biopsies are somewhat overprocessed, but 90-95% of the time
they look perfectly fine. We can have GI biopsies in a case th
Hi Peter,
We need more info:
1. What is the tissue(s)? Species?
2. What is the marker(s) in question?
3. Answer 1 and 2 with regard to the shared images?
4. What is the expected staining reaction?
5. Is there a control section placed on the same slide? If yes...
6. How does the c
Honestly, I don't see the point of using this instrument in the histology
lab (at least not my lab as we are set up). It would seem odd that we are
being "ultra safe" when dispensing the formalin but then it's kind of the
"wild west" when we have to discard the previously archived wet tissue!
Obvi
Hi Nancy,
All routine specimens in our hospital are placed in formalin in the OR.
Breast lumps and mastectomy specimens are sent up fresh so that they arrive
STAT (to minimize cold ischemic times) and lymph nodes for lymphoma
protocol are also sent up fresh. [*they actually send sentinel nodes up
f
Background:
I read on a vendor website that tissues stained using In situ hybridization
should not exceed 32 hours fixation. Greater than 32 hours can produce
false negative results because the validated retrieving protocol can be
inadequate.
We in fact have seen evidence of this in our lab (mysti
This is more of a survey than a question:
For those of you tracking and documenting your cold ischemic times for
breast tissue (ie time out of body to time sliced [as needed] and immersed
in formalin), and I assume most of you are...*what is your average time?*
*Background:*
*I ask because my dir
Hello experts!
I would like to find out what others have done to validate a new cryostat.
I believe it would basically be the same as validating a new microtome
except specimens (ie blocks) are plentiful for microtomy whereas for us,
fresh tissue for cryosectioning is not readily available.
Backgr
Hi Bernice,
In my lab, water under the sections is unique to charged slides. And you
are correct, if there is water under the section when the slides are heated
for antigen retrieval, the boiling (or at least very hot) water will damage
or entirely destroy the section.
We allow the charged slides
Hi martha,
Prognostic markers must be re-validated (Eg.s Breast markers and CD117) as
you described.
Every Ab in your menu should be tested (as you would for a new a new lot)
and do not forget to validate your H&E (with various tissue types) and SS
as well. For the H&Es, if possible do side-by-sid
Hi Tim,
The usual source of bubbles is a deteriorating syringe or lines or valve.
Have you observed the syringe while running the Clean Fluidics maintenance
function? There can be some "micro" bubbles in the syringe on first draw of
a reagent however, if working properly these tiny bubbles quickly
Garratt wrote:
> Evaluation of Virus Inactivation by Formaldehyde to Enhance Biosafety of
> Diagnostic Electron Microscopy
>
>
> https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4353909/
>
>
> It is nice to have a reference.
>
>
>
> John
>
> On Thu, Aug 6,
Hi Amy,
Formalin fixed tissue is no longer infectious...unless you are talking
about prions (eg scrapie, BSE, etc). So there should otherwise be no
concerns or additional precautions required.
Cheers,
Greg
--
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A 1P0
*Everything in moderat
Hi Wanda,
If you have just purchased the Spectra stainer recently (ie still under the
first year warranty), demand that Leica send their Application Specialist
back on site. if they promised 2000 slides with no drop in quality over
that period (I believe it is 2000 slides or 7 days, whichever come
Hi Charles,
If memory serves, there is a thermal fuse or switch that will shut the unit
down if it exceeds a certain temp. Once the unit cools, the switch is
reset. So either that switch needs to be replaced or there is a problem
elsewhere that is causing the unit to overheat. Replacing that switch
Nancy,
You used the word “blot” in your reply to Charles, and while I can’t be
certain that you did not mean the same...I would like say in the interest
of clarity, that it is safer to carefully “wick” the water away from under
each section prior to baking. Blotting (like we might do after a Gram
Hi Charles,
It adhesion problems can arise from several sources, some have already been
mentioned. Here are two that I have experienced:
1) Bad slides. Either a manufacturing defect such that the positive charge
is insufficient or the slides were somehow compromised after opening them
on the bench
Hi Sandra,
>From your post, it looks to me like you are assuming the recommended
dilution should work. I apologize in advance if I have misinterpreted your
post in this regard.
When starting with a new antibody (including a new vendor for same Ab
and/or a new clone), do a range of dilutions that i
ece of tonsil kept in
> formalin before processing? Could over fixation be the issue?
> I have found that maintaining tight control of control tissues is
> important which includes minimum and maximum fixation time.
>
> John
>
> Sent from ProtonMail Mobile
>
>
> On Tu
tralia
> Locked Bag 4001, Westmead 2145, NSW Australia
>
> ♲ Please consider the environment before printing this email.
>
> -Original Message-
> From: Greg Dobbin via Histonet [mailto:histonet@lists.utsouthwestern.edu]
> Sent: Wednesday, 8 May 2019 5:07 AM
> To:
Hello colleagues,
I recently stained (IHC) a section of normal tonsil from another facility
with p16 and the resulting stain was better than the same stain on a
section of my labs own normal tonsil control.
This has led us to question our processing schedule. I am not concerned
with our fixation
Hello all,
While I do not disagree with anything that has been said thus far regarding
the failed refrigerator and moving forward, I think Tim Morken's
information and shared experience was the most relevant! I know that my
Bond instruments are running 24 hrs a day, so some of the more commonly
ord
Hi Karen,
As mentioned by others "decay" is not likely going to be an issue. More
concerning for you could be not knowing how those tissues were handled
prior to processing 10 years ago.
Presumably, you now track cold ischemic times and have standardized your
fixation protocols for breast tissues
Hi Curt,
You can de-stain the hematoxylin counter stain and then do H&E and all
should be fine. Subsequent IHC is possible but obviously you would need a
different color chromogen to differentiate the new stain from the previous
one.
There may be a problem getting the section stay on through a se
Hi Dr. Cartun,
It has been many years since I worked in EM but I my recollection is that
tissues could remain in 2% Glut indefinitely without detriment (for EM
purposes). However, Osmium tetroxide had to have a limited exposure.
Greg
--
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A
Hi Linda,
Is this a problem that has just started or has the red kit always had this
artefact?
- If it just started, perhaps it is a problem with a particular kit lot
number (or numbers). If so, get Leica Tech service involved to see if
anyone else has reported a problem.
- If it has
1:100 with ER-2 for 30 mins.
Clone is the V600E from Spring BioScience
Looks great for uis and we are performing well on EQA surveys.
Greg
--
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A 1P0
*Everything in moderation...even moderation itself**!*
__
Yes, it works. Very well actually. And we have done well on both a cIQc and
a CAP survey.
Greg
On Tue, Jul 24, 2018 at 3:20 PM Alminawi, Samira <
samira.almin...@sunnybrook.ca> wrote:
> Hi Greg,
>
> How is the quality of the staining? Is it working for melanoma?
>
> Samira
>
>
>
> *From:* Greg Do
Here in Charlottetown we are using V600E (VE-1) clone (Monoclonal) from
Spring Diagnostics. We run it on the Bond platforms at 1:100 with ER-2
(high pH) for 30 mins.
Greg
--
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A 1P0
*Everything in moderation...even moderation itself**!*
__
Pass The General MLT Certification exam, as our MLT exam includes Histology
and Microbiology. :-)
Greg
--
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A 1P0
*Everything in moderation...even moderation itself**!*
___
Histonet mailing list
Hello all,
I would like to find out what other clinical path labs are doing in Canada
in particular but in the US as well with regard to retention of blocks and
slides. Are other labs filing pediatric cases separately from adult cases?
I am required to archive specimens from pediatric cases separa
Hi Charles,
If you are using the Leica Diluting Buffer or another commercially prepared
diluting buffer the antibody should remain stable for a very long time. I
can't say how long, I suspect there would be numerous variables to be
considered but generally...I would expect very little loss of inten
Hi Laurie,
We are purchasing the antibody only from Roche/Ventana and optimizing it
for use with our detection system (Bond Refine Detection kit) and the
Bond-III stainer. Ask your Ventana person about buying the Ab only.
Greg
--
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A 1P0
*
Hi Merissa,
As far as I am aware, insufficient paraffin infiltration would only affect
sectioning. The epitopes that we we are attempting to stain with IHC are
affected by pre-analytic factors such as fixation, cold ischemic time and
perhaps heat (too much) but plus or minus wax should not be an is
Hi Lauren,
Further to Tony's response...there are also thermal fuses that shut the
power off to a drawer if the power (or temp IDK which) as a safety feature.
So quite often we find one warming drawer or the other can be reset by
powering the unit down for a couple of hours. It normally works fine
Hi Paula,
Let me first say I am Canadian and my lab is not governed by CLIA or CAP.
But for what it is worth, here are my thoughts...
When thinking about validating antibodies for IHC you must first consider
whether the antibody in question is Class-I (prognostic eg Breast markers,
CD117, etc) or
No pretreatments for anything that is not formalin fixed. I think 95% ETOH
for a fixative but but others may have a better idea than I.
Greg
--
*Greg Dobbin*
1205 Pleasant Grove Rd
RR#2 York,
PE C0A 1P0
*Everything in moderation...even moderation itself**!*
Good day colleagues,
We are occasionally being asked to do an acid-fast stain (Ziehl-Neelsen) on
a alcohol-fixed cytospin preparation from a cytology fluid.
Typically in Cytology, if a specimen is very bloody, we will add acetic
acid to lyse the red cells. Does anyone know if acetic acid would hav
Hi Folks,
As it happens, we had a lymphoma case this week that was Kappa restricted
(confirmed at a consulting laboratory by Flow Cytometry) and our Kappa and
Lambda ISH staining was negative for both! In discussing it further with
our Hematopathologist, our ISH probes for Kappa and Lambda in bone
Hello Renee,
We do K & L by ISH on the Bond-III and BondMax with Refine detection
without issue on any of the tissues we have run (GI, skin, BM, LN). We have
a normal tonsil control section on every slide. Do you also run a positive
control on every slide? How does it perform? Next step would be fo
Hi Folks,
Can some of you share what your practices are regarding storage of blanks
for possible subsequent IHC orders?
Background: We cut a blank on every prostate core in case a PIN4 is ordered
after the H&Es have been read. So each week, we have 72-100 blank slides to
store. Space is an issue e
Hi Folks,
I haven't been on here much lately but it is nice to know that we are all
here for each other when needed!
My problem is section adherence during IHC staining. And it is not all of
the time it is intermittent; so not all of the time and not on all slides
when it does occur.
Background:
Vanessa,
We strongly encourage people that make such requests to use a funeral home
or crematorium to handle the tissue for them (for example an aborted fetus;
or some cultures wish to have an amputated limb buried in their future
burial plot). If they take their own amputated limb and then were to
I can't speak to the the T-cell receptor sites but the only option for
getting decent sections at this point is option number two. In the future,
(if there applicable) the cut surface of a cryosectioned block can be
recovered with OCT, frozen, and stored in an air-tight whirl-pak or ziplock
bag in
Joanne,
I do not think you should expect to get perfectly uniform IHC staining in
breast core biopsies every time. At our hospital, we prefer to avoid doing
hormone receptor stains on breast cores because they are not as "robust" as
a lumpectomy or mastectomy specimen.
The unevenness could be due
Judith,
The red chromagen is suseptible to "fading" if stained slides are left in
either alcohol or xylene for too long. Check to make sure that everyone
involved is following the same procedure for the dehydraytion, clearing and
mounting of the stained slides.
Greg
--
*Greg Dobbin*
1205 Pleasant
Buy yourself a roll of WHMIS labels. Get ones that are sufficiently large
to be easily read while still fitting on the side of the secondary
container. All necessary info will be captured there once you check the
right boxes, etc.
Cheers,
Greg
--
Greg Dobbin, R.T.
Chief Technologist,
Anatomic Pat
Are you hoping to measure competency or productivity?
If it is competency that you hope to assess, allow your tech to cut blocks
of a variety of specimen types for 1 hour *without* distractions. If they
cut 30 or more blocks and produce good quality sections, then they are
meeting your standard.
Hi Folks,
I am having difficulty finding a fine mesh biopsy cassette that is
compatible with my TBS cassette printer (i.e. *45 degree angle* print
surface). The holes in the TBS cassette that I am currently using are too
big (~1mm). Can anyone suggest a supplier with a good quality cassette that
wi
57 matches
Mail list logo
