Amos, hello. Do you have a reference for this? All my files talk about
"endogenous" B-gal in kidney and pancreas and other organs (but then article
talks about after lacZ transgenic manipulation) or demonstration of
alpha-galactosidase in kidneys or in senescence associated or lysozomal storag
Anna,
Don't know if you are talking human or non-human control or if makes a
difference and I don't want to get into that whole discussion again. But if a
mouse control is OK, one of the cleanest and nicest systems I used for B-gal
was to get a transgenic mouse, easily obtainable with a Tie-2/
I got your message Gayle (through Histonet) although haven't heard much at all
weekend otherwise. Use IE.
Ray in Lake Forest Park, WA
- Original Message -
From: "Gayle Callis"
To: "Histonet"
Sent: Monday, May 4, 2015 3:21:01 PM
Subject: [Histonet] Can't log into Histonet to do a
Hi,
Thanks for info and happy to hear that. Didn't realize. May 10-15 is Worlds
largest International Science and Engineering Fair in Pittsburgh this year and
maybe will see others there and watch a high school histology project from
somewhere bring home a $75,000 scholarship.
Ray
The following has to do with histology and STEM (science, technology,
engineering, math) so if not interested, please ignore. But I believe it can
have real meaning to the profession of histology at the NSH, state society and
local levels.
I am elected to the Board of WSSEF (Washington Stat
Hello Garrey,
Curious myself, CAP contact info seems to be greyed out on website unless I
officially log in and for now my concerns are with the Washington State Science
and Engineering Fair for K-12 and golf game.
(1) There are at least two phrases in the ANP.21450 which could be parsed out
I asked about this in a different vein months ago. Has anyone shown a
strawberry or ground meat or slim jim or orange peel as a bacteria/fungus
control used for diagnostics to an inspector inspecting the lab and was there
any comment from the inspector either positive or negative. Never heard b
Hi all, as usual Gayle was right on. Use a buffered (more gentle) formic acid;
not just formic acid per se of any water diluted concentration. For end point
testing, critical, we used a radiograph machine instead of chemical endpoints
which is also fine;we just had access to a lot of equipment
Debra,
it appears most of the histology world disagrees with me but I stand by my
post. If TRAP didn't work with formic acid in our hands, a major
pharmaceutical treatment wouldn't be ready to help women with post-menopausal
osteoporosis.
Ray
- Original Message -
From: "Sarah Mack"
Hi Debra,
My experience differs from Elizabeth and others. Indeed have made thousands of
mouse bone preparations with formic acid decaled sections. Here is an article
that didn't copy paste so well:
RANK is the intrinsic hematopoietic cell surface
receptor that controls osteoclastogene
Apparently there are numerous interesting ways for fungus or bacteria controls
to be had from orange peels to hamburger to slim Jim's to hot dogs to
strawberries to . Sounds like fun to me. I'm curious, with the emphasis
now on quality control in labs run amok, has anyone passed a rigorous
I am not disagreeing nor am I sticking up for the company and not sure I'd even
agree with the company but I think there is much more to this, from what little
I know of them, than just mixing up two specimens at a physicians office. I
believe their point of view of the company is besides patie
Michael,
Not sure from your explanation if your mCherry tag is less intense on the
inside of sections or if you are referring to the P and RFP Alexa and Cy5 stain
intensity If the former, not sure about that. If the later, having done
very similar thing in grad school, but with different
Kristopher,
beta-gal is an enzyme and as far as I know is rendered inert after FFPE. I did
do fresh skin in X-gal to target, as you would with any whole mount specimen,
THEN FFPE and cut sections to it to see the signal. If that is not an option,
go after it with an anti-beta galactosidase an
Tiffany,
Have used 10%NBF on muscles but also alcoholic fixatives -alcoholic formalin or
absolute- just always preferred 10%NBF since it gave the morphology and
counterstaining I wanted. diastase in a 6.0pH buffer (don't heat above 40
degrees if trying to speed up heating-kill the diastase) an
Oh, oh, sorry. I goofed. I thought this "double labeling" was technical and
not regulatory.
Ray
Lake Forest Park
- Original Message -
From: "Timothy Morken"
To: "Neelam Joshi" ,
histonet@lists.utsouthwestern.edu
Sent: Thursday, October 23, 2014 7:23:58 AM
Subject: [Histonet] RE:
Neelam,
If you are referring to immunolabelling for transmission electron microscopy,
it is relatively simple to apply and this worked wonderfully for projects we
did, to use 2 antibodies to two different epitopes but each antibody with a
different size gold particle attached. The gold particl
Hi Joyce,
Absolutely agree with recycling concept, value, money saved and no fumes in lab
(if using newer models) and if used properly. I've always been curious about
the concept of a lab recycler making xylene "purer" by distilling out isomers.
Which unit do you have? meta-xylene is in grea
Heartbreakingly sad,
I do not know where the current regulations are but safety, as Terri rightly
pointed out, is an accident that did happen. Not an anecdote, you can look up
March 1985, Jackson Memorial Hospital in Miami (years after I left).
Patient went to surgery, had some cerebrospina
Alpha Histotech,
wanted to be sure that I did NOT tell you to drop everything in life to look to
research exclusively. So I cut and pasted this from my original message
"Research histology should not be overlooked
I stand by that statement.
I agree with Emily that funding in research is
Alpha Histotech-
I'll put in my few words even though I'm not active anymore and possibly from
different perspective. But also using a few assumptions and if my assumptions
are wrong then the rest of what I say is probably meaningless. Not IDíng your
e-mail address but if you've worked 3 j
Eva,
Not sure you are going to find that much "published" on HA-tag antibodies.
That 9 amino acid sequence used by most to tag; people are looking to identify
the fusion partner of the tag and worry little about publishing on the tag
itself. Same with the famous 8 amino acid sequence FLAG-tag
I got a bounce a week ago and contacted Linda M.in private and was deemed
fairly innocuous but as with all such things even remotely suspicious I deleted
it and never, ever look into such things. As it turns out, was talking a few
days ago to a judge for my district STEM science fair who happe
Hi Merissa,
don't know if you got any private idea responses so I'll throw in my opinion.
I would always worry about some of the things you are mentioning and that are
standard thoughts regarding biotin block, retrieval, etc in IHC.
But I would think about your serum, which I steadfastly a
Hello everyone, thought I'd chime in here as I just returned from helping and
judging at the 2-day WSSEF (Washington State Science and Engineering Fair) in
Bremerton, Washington. Had 500 incredible projects from all over the state.
It is our state fair that leads into the ISEF, giant Intel sci
Hello, I agree anti-human CD68 might not be best for mouse tissue but there are
specific mouse CD68 (the mouse homolog to human CD68) antibodies out there.
FA-11 is one of my favorites. Have used both F4/80 and (rat anti-mouse CD68)
on flow, frozens and FFPE. Anna, a literature search gives y
Hi Patty,
You sort of piqued my curiosity of what is going on in the zebra fish world and
found this youtube, 6 minute film
http://www.youtube.com/watch?v=kZDwo20hl1E&feature=youtu.be
About what is going on at Welcome Trust Research on zebra fish but maybe you
know all this already. Anyway,
Patty,
did some zebrafish work years ago, either pure histology or sections after we
did whole mount ISH on the embryo's. As per Jack Ratliff's post, paraffin I
found was really tough for orientation and for getting enough sections of such
small embryo's. But would use, as Jack suggested, JB-
Hello all out there,
This is regarding: Ray Koelling; currently from just north of the Seattle, WA
area. If you and I have connected in some way over the last 47 years, the
following is a message concerning my (semi)-retirement that I hope you will
enjoy and anyone with little or no interes
Hugh, I agree with Rene about the possibilities and yes a mask might be
overkill. But far, far, far more than that for me, the description of this
"research" as stated bothers me greatly. I am surely no expert on this
particular line of research but it is my understanding that projects looking
I recently had the same question. They were gobbled. Try Endogen antibodies at
Thermo Scientific conglomerate.
Ray
Seattle, WA
- Original Message -
From: "Ronald Houston"
To: histonet@lists.utsouthwestern.edu
Sent: Friday, October 4, 2013 8:16:31 AM
Subject: [Histonet] Endogen
D
Just hours after I sent out my recent e-mail regarding the digital and internet
conundrum and how uncivil it can tend to make humans by its indiscriminate use
without first any thought, here is a headline copy/paste from the national
news:
Police: Girl, 12, Bullied Online Before Suicide
I
I absolutely agree with "well said" and I know I'm going to just hate myself in
the morning for doing this but I can't stop. I hope everyone (I've mentioned
the book to our school board and every kid I mentor in biotechnology and tutor
in math) will get "The Dumbest Generation:How the digital
Could I ask exactly what you are looking for in mouse femur and human tonsil
the tonsil I assume is a control? For what? Whatever you are looking for is
obviously in human tonsil. Are you positive it is in murine femur. I used to
stain for 15-20 targets in EDTA decaled mouse femurs but my
Betsy,
Don't know if applicable to this situation but in the past I have on occasion
in other labs noted some stain going off and after checking everything, looked
at the tap water. In fact I backtracked through the water utility once to see
where the water came from, was delivered and what the
Victor, I completely agree with Tony and Renee's methodology assessment but I
would also ask you to look at your induced chondrogenesis model (I've never
done this but have done many, many other cell culture models). If you go to
this paper http://biotech.korea.ac.kr/bk21/bbs/upload/bk21bbs_
Marina,
I agree with Paula Pierce, have used the same anti- GFP antibody on some
projects but at 1:500-1:1000 area and with a much longer 8-15 minute retrieval
in citrate. Also, I know you know to be aware of "how much" the mice are
transfected . In my transfected mice (NOT GFP and in a form
I whole-heartedly agree with and applaud Amos Brooks playing Devil's advocate.
I certainly would never discount the degree of danger with what is a high
explosive and would take all due caution using disposal people who know what
they are doing. But there is a use for the substance and one needs
Great and fantastically said and to answer your last question in my opinion
"No!!". My original point when this started.
Ray
Seattle
- Original Message -
From: "WILLIAM DESALVO"
To: koelli...@comcast.net, akbitt...@geisinger.edu
Cc: "histonet"
Sent: Tuesday, January 31, 20
Fascinating... and that gets me back to my original ponderance. Why all the
"false" histotechs? Are there people trying to sneak into flow cytometry who
have never run a flow cytometer or clinical chemistry medtechs who have never
sat in front of an analyzer or cytotechs who don't know an epithe
I hope there is at least a bit more to clinical histology than quantity and
speed. Maybe I know the particular high schooler you mentioned who was so good
since I mentored that Mercer program/class for years and years and even brought
in class-wide IHC hands-on projects. Before the teacher left
Lynn, Perhaps the most provocative and ingenious idea yet. Have your
"competency to cut" with you; like many other files, a drivers license or
others for example, follow you through (working histology) life.
Excellent!
Ray
Seattle, WA
- Original Message -
From: "Lynn Dike"
Or as Gayle wisely pointed out it might be interview sectioning to
differentiate those who "cut out" on an interview.
While there is no right or wrong to this question, I'm still not convinced that
it is a useful tool for you or HR to just have a routine "can cut (section) on
rotary mi
Not upset in the least. Just posting my own questions and doubts within the
parameters of the situation. When the Chinese philosopher who fell asleep under
a tree and dreamt he was a butterfly and then spent the rest of his life
"asking" if he was a human who fell asleep under a tree and dreamt
This is certainly an interesting thread and I generally hate to get into these
ever but I still can't figure out one thing and never have over all these years
in pathology. What other endeavor in life and job seeking is an on-the-spot
demo that you can do something required at a job interview? D
Carl's choice is excellent. The classical B-cell marker might be CD20, and does
work great, but as with all antibodies, you have to be aware of exactly what
you are after. CD20 comes on line after CD19 induction so CD20 might not be
seen on very early b-cells. Like wise CD79a is accompanied by
Hi Lydia,
I think Tony Henwood has it exactly right in talking of DAB intensification.
The article he sites and several others show how much more sensitive DAB
intensification can make an ordinary iron reaction. And you are not looking in
bone marrow or spleen but somewhere where there is
Cheryl,
Have done some things with ordinary sectioning cryo-electron microscopy but not
your specific application. But can go to Cold Harbor Springs Protocols and
they can get "Immersion Freezing of Cell Monolayers for Cryo-Electron
Tomongraphy".
Even better (more immediate) go h
Adam/his fellow grad student,
I could be somewhat off not being up to date with current literature. Sorry.
But used to look for Flk2/Flt3/CD135 in murine bone marrow and not the bone
itself. In immature hematopoeitic progenitor cells. Along with looking in
thymus, etc. But for somet
Hello Per,
Have no particular interest in neuronal retrograde tracing and have certainly
never used anything for that. Just general scientific curiosity and a few
minutes time awaiting a home project. Found Fast Blue (synonym: Diamidino
compound 253/50) on Sigma website cat # F5756. Good lu
Allison/Toni,
Thought I'd throw this out. Maybe nonsense. If you have such acidic tap water,
could there be heavy metals lead, magnesium and others (acid tap water does
that) in your tap water rinse from being leached out upstream. William DeSalvo
talked about the quality of tap water fluctuati
Hello,
Maybe I'm remembering wrong since I deleted the original message of Bretts
looking for murine CD20 on mouse FFPE (hopefully rabbit origin?). If so
anti-murine CD20 that can be made to work on mouse FFPE tissues certainly
exist. Have used them from BD (flow reagent), Santa Cruz and ot
Hello,
It is very inexpensive, a few dollars, to buy a counter-top ice crusher, that a
professional bartender might use to crush ice for drinks. Can be bought at
most any store. We crush ice every day on a daily basis to create a small tub
of ice we need for reagents. Only the size of li
Amy,
In my opinion the answer is, Yes, certainly possible. Also in my opinion the
answer is No, probably impossible. Even with a Yahoo address I'm assuming that
this is a research/biotech model and project. In a previous life, when quietly
gagged, I went about this on numerable occassion
Hi Gayle,
I've used this extensively. But never from BD. Was too simple to make in house
and then we had control over tweaking it if needed. Made it up per the formula
you wrote and with all your caveats and warnings with which I completely agree.
We loved it for certain things. For instance,
In a previous lab, we went through that. In order to keep the large nitrogen
tank in a small room that had 2 access doors, there had to be an oxygen
monitoring inside the access points. Oxygen level had to be at some minimal
level otherwise an alarm would sound. Just in case, with doors closed,
Hi Jerry,
I would never try to persuade anyone. I'm no smarter than the next lab worker
trying to make sense of this and science biology in general. But this is how I
see TUNEL and FFPE and after all the years I'm happy with things.
I have not noticed false positives at all when looking
Tyrone,
I agree with Amos Brooks about the Chemicon kit and I agree with Jason Palmer
about about Promega, especially in regards to smaller, controlled volumes to
decease cost per slide. In addition, I only used both those kits directions as
starting points. Took care of the proteinase dig
The hamster isotypes are tricky and ill-defined. Some people (and me formerly)
use secondary cocktails. Sometimes Armenian and Syrian are interchangeable.
Sometimes they are completely not. BD biosciences has a great Hamster Ig chart
that you can use for flow (or IHC) reference.
Ray Koelling
Jennifer,
In the past, upon occasion when need arose, I'd use directly conjugated primary
antibodies. Mainly biotin or dig or peroxidase but also FITC directly to
primary. Then can look at it fluorescently or come back with an anti-FITC of
some kind (several are good). Used these for my
Hi in grad school taking microanatomy and pathology classes, 2 that I heard are
this: The surface area of all the alveoli in the lungs of an adult is between
40-70 square meters. That seems reasonable in having a 40-70 square meter
surface (where all gas exchange takes place) represent all th
Joost,
I was curious myself what was out there and maybe someone has answered you but
I haven't seen anything on the HistoNet. Haven't done anything with murine
CD86 (B7.2) for 3-4 years but this is what I can recall. Of course you know
the whole CD80/86/CD28/CTLA-4 costimulatory story is
DONGTAO FU,
Was hoping to see some response for curiosity sake but haven't so this is my
take. In a former life have seen such an occurence. Not with these same
reagents but similar happenings. And while I didn't take the time to
investigate exactly why, we attributed it to posttranslatio
Isn't a Klatskin tumor (of the biliary tree) spelled with A and not O? There
are multiple pictures and references to Klatskin's Trichrome. Try A spelling
and not O.
Ray Koelling
Research Pathology
PhenoPath Labs
Seattle
-- Original message --
From: Pat Laurie
TF and Emily,
You can use antibodies designed for Western blots. They can work. In a
western blot, a lot of times these are run under denaturing conditions (a type
of soap is used to break the secondary and tertiary structure) and so the
epitope seen is linear. That antibody for Western's wo
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