[Histonet] aniline oil/Holzer stain

[Susan Bachus via Histonet]

I'm trying to locate aniline oil for the Holzer glial fiber stain.  I 
purchased what was listed as aniline oil from ENG, but the first hint that 
something was wrong was that it was incompatible with the chloroform also 
included in the differentiating solution.  Looking at the fine print on the 
bottle, I saw that it was actually "aniline oil, 2.5% aqueous", which I 
would have interpreted as only containing 2.5% water, except that it seemed 
to be completely aqueous (2.5% aniline?), inasmuch as it wouldn't mix with 
the chloroform.  I can't find any other source online.  I see that it is 
also called aminobenzine or phenylamine, but am also having trouble locating 
sources under those names.  I realize aniline oil is relatively toxic, but 
am using it under a fume hood.  I tried dipping slides separately in the 
(apparently aqueous) aniline and then chloroform, but still didn't get good 
differentiation.  Can anyone suggest a source or a substitute?  Many thanks 
for your help!   Susan 



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Re: [Histonet] looking for The Art of Histology by Stephanie De Ritis, Sept. 6, 2004, Advance

[Susan Bachus via Histonet]

Sorry, Freudian slip, of course I meant Advance for Medical LABORATORY 
Professionals!   Sometimes my "hats" get mixed up...   thanks,  Susan


-Original Message----- 
From: Susan Bachus via Histonet

Sent: Saturday, September 03, 2016 1:12 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] looking for The Art of Histology by Stephanie De Ritis, 
Sept. 6, 2004, Advance


I'm trying to find a pdf of this paper for my students.  I have an ancient
battered xerox copy in shades of gray.  For some strange reason it's not in
the Advance Medical Library Professionals online archives.  I'm hoping that
Stephanie is in Histonet, or that someone else has access to a copy.  My
students thank you!   gratefully, Susan


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[Histonet] looking for The Art of Histology by Stephanie De Ritis, Sept. 6, 2004, Advance

[Susan Bachus via Histonet]

I'm trying to find a pdf of this paper for my students.  I have an ancient 
battered xerox copy in shades of gray.  For some strange reason it's not in 
the Advance Medical Library Professionals online archives.  I'm hoping that 
Stephanie is in Histonet, or that someone else has access to a copy.  My 
students thank you!   gratefully, Susan 



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Re: [Histonet] RE: Brass cryostat chucks

[Susan Bachus]

You got me to thinking, that I would be in a real jam if I ever lost mine! 
I'd rather buy one from someone who has old ones they don't need any more, 
but I did find some (at least I think this is what you're describing) 
available online at:

http://www.psl-equip.com/shop/advanced_search_result.php?keywords=CC-3330+&osCsid=683519442648223efdbb7a61f87d1c05
(You might have to "register" to see the information, but the part # is 
CC-3330 & they cost $22.)


-Original Message- 
From: Maryott, Bridget

Sent: Wednesday, November 23, 2011 9:27 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] RE: Brass cryostat chucks

I know I just saw a box of these in my garage. I will check over the 
weekend. I have a plethora of old microscope/microtome parts left over from 
my father that I have no clue what to do with anymore.


-Bridget Maryott, HT (ASCP)
 Tissue Sample Management
 Ventana Medical Systems, Inc.
 a member of the Roche Group
 1910 E. Innovation Park Drive
 Tucson, Arizona  85755

 Tel: 520.229.4022
  Lab: 520.877.7145
 mailto: bridget.mary...@ventana.roche.com


Message: 9
Date: Thu, 17 Nov 2011 15:37:03 -0500
From: "Goodwin, Diana" 
Subject: [Histonet] Brass cryostat chucks
To: "'histonet@lists.utsouthwestern.edu'"

Message-ID:
<09411E0112A96A459D8D5FBDAB9C15C72480DAE2DF@HAMEXMBA.rwjham.local>
Content-Type: text/plain; charset="us-ascii"

Greeting, Histonetters.

I am in search of those brass cryostat chucks with the holes in them.  Any 
ideas?  Googled my brains out with no luck.



Diana G. Goodwin, BS, HT(ASCP)QIHC

Department of Pathology

Robert Wood Johnson University Hospital at Hamilton

One Hamilton Health Place

Hamilton, NJ  08690

Ph:  609.631.6996

Email:  dgood...@rwjuhh.edu




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Re: [Histonet] calibration of pH meter

[Susan Bachus]

This may sound simplistic, but it works for us:   whenever I need to use pH 
something, I quickly first dunk the electrode in the buffers above & below 
what I'm pHing (rinsing after each dunk, of course)--if the meter reads 
accurately I proceed to measure, if it's off by more than a few % I 
recalibrate before measuring.It's true that a new electrode can stay 
well calibrated for long periods.Susan
- Original Message - 
From: "Jan Shivers" 
To: "histonet" ; "Akemi Allison" 


Sent: Tuesday, February 01, 2011 1:36 PM
Subject: Re: [Histonet] calibration of pH meter


We calibrate our pH meter every day that it's used, which can be daily 
during heavy workload weeks, using multiple calibration buffers.


Jan Shivers
UMN Vet Diag Lab

- Original Message - 
From: "Akemi Allison" 

To: "histonet" 
Sent: Tuesday, February 01, 2011 7:54 AM
Subject: [Histonet] calibration of pH meter



Good morning everyone,

I am curious how frequently you all calibrate your pH meter, 
particularly when you make up solutions for muscle bx's?  Do you do  it 
daily, weekly, or monthly?  I am trying to come up with an  acceptable 
standard in a clinical histology lab, verses a biotech  lab, which 
makes-up large quantities of reagents on a daily basis.


Thank you in advance for your information,
Akemi


Akemi Allison BS, HT (ASCP) HTL
Director
Phoenix Lab Consulting
Tele: 408.335.9994
E-Mail: akemiat3...@yahoo.com

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[Histonet] Can anyone put an old Mettler microbalance to good use?

[Susan Bachus]

I have an old surplus Mettler balance that is no longer worth recalibrating--I 
hate to waste anything & am wondering if anyone might find it useful enough for 
a display or something like that to be worth the cost of shipping?
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[Histonet] in situ hybridization histochemistry question about radioisotope

[Susan Bachus]

I have routinely seen that we obtain slightly different results (like 10-20% 
variance) in using 35S dATP for TdT-catalyzed tailing reactions of 
oligonucleotides from month to month as we use different batches (made fresh 
monthly) of the 35S dATP.   But on a few occasions over the years we have 
found dramatically reduced results with a particular lot.  This happened 
again in February, and I confirmed by comparing directly between the Feb. & 
March lots in March that the results with the Feb. lot were half that of the 
March batch (of course, given the half-life of 35S, they would be expected 
to be down some, but not by half!).  The company was good about believing me 
and not charging us for the March "replacement" batch, but they have no idea 
what could cause this to happen occasionally.  They check the specific 
activity, pH, purity, etc. and run a "bio-assay" that is a binding assay 
each month, but were not set up to test the enzymatic reaction.   I told 
them that years ago when this happened it was finally determined that the 
DTT concentration was unusually high and interfered with the reaction, but 
they discounted this possibility.  I guess I should just count my blessings 
that things are working again now (mercifully, I was able to cling to 
optimism, based on past experience, that the isotope was the problem, until 
I could confirm this myself), but it's very frustrating that the company is 
not motivated to check the efficacy of the item for this particular 
application (which I would imagine a lot of customers use it for) each 
month, or to figure out what causes this to happen on occasion (so as to 
avoid it!)--we lose a lot of time and other expensive reagents (e.g. the 
TdT) each time this happens.   I also don't know how to interpret it--I know 
that as the enzyme begins to degrade it just means that the tails are 
shorter, which I can compensate for with a longer film exposure, but I have 
no idea what's going on when the isotope causes the problem & am afraid to 
use those batches.  Does anyone else using ISH have any ideas about what 
causes this variability?   grateful for any thoughts,   Susan 

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Re: [Histonet] Histo Stories

[Susan Bachus]

Sorry to be so late to the party, hope I'm not too late to share my 
story:   I still vividly remember being shown a "career documentary" film 
about histotechnology, in junior high school, back in the 60's, by my 
biology teacher, Lynda McCurdy Ballingall--the lovely lady who influenced my 
life more than anyone else, and who I just enjoyed the privilege of visiting 
in Chicago, where she lives now, when the Society for Neuroscience meeting 
was held there last year.   (She swears she remembers me staying after class 
to continue drawing what we were observing under the microscope, though I 
have absolutely no recollection of this!)
But I didn't think much more about it till I earned my PhD in 
Psychobiology, decades later.   Of course in psychobiology research 
histology is an important method, to verify lesion damage, electrode and 
cannula placements, etc.   Back in the "good old days" a psychobiology lab 
supported a technician, an animal caretaker, a secretary, and a histologist, 
in addition to grad students and the primary investigator.   I began as that 
golden era was waning though, at least in our lab.   I survived by teaching 
undergrads, and the histologist was being "let go".   Just before she left, 
our delightful histologist, Mrs. Anne Madsen,  patiently trained me, and I 
then trained the students who came after me.   The only drawback was that 
she was so expert that everything worked perfectly in her hands.  It then 
took me the next few decades to painfully learn by trial & error how to 
troubleshoot as things gradually went wrong (pH's drifted off, gelatin 
congealed, etc. etc.).
I gradually added other related methods to my armamentarium, like 
immunohistochemistry and in situ hybridization histochemistry, while a 
post-doc at NIH.  Not least of the "goods" that I reaped at NIH were the 
supplies that labs discarded when they moved, which I hoarded in my basement 
at home in anticipation of setting up a lab of my own in academia, where 
funds don't flow as freely as those at NIH, someday--enough, it turned out, 
to set up my own functional lab nearby at George Mason University!
So when I began teaching a graduate level Histology/Histochemistry 
course that I kluged together on my own at GMU (thank goodness for Carson's 
textbook!), several years ago, I made a big deal of teaching my graduate 
students the nitty-gritty "nuts & bolts" theories/rationales underlying the 
methods, to prepare them for troubleshooting in their own futures!  Some of 
you may recall that one of my students inadvertently stirred up a flurry of 
debate about the purpose of Histonet a few years ago when she asked a 
question that was misinterpreted as "cheating" while utilizing Histonet as a 
"resource" (not what I had in mind when I encouraged them to subscribe to 
Histonet!) in preparing for a lab that embedded a "mystery" to solve in the 
exercise of learning how to use Nissl stains (trying to make things more 
interesting).
One downside to having learned "on the scene", as it were, is that I 
never received formal training or licensing in histology.  So now that I 
find myself unable to afford to continue indulging in teaching as an adjunct 
($3K for a 5 credit class), since my husband's death, I also find myself 
excluded from "real jobs" in histology.   (If anyone in the DC/Northern 
Virginia area could use someone like me, please email me!)
I laughed a lot over others' stories of their kids' exposure to their 
work.  Mine were also  practically raised in labs (my husband was also a 
scientist [electrophysiologist]).   They have more vivid memories of the 
animals we used in our psychopharmacology research though--to this day (my 
son is graduating from college now) they proudly profess to never having 
felt the slightest temptation to abuse drugs (thank goodness!), having been 
appalled by the pathetic bedraggled appearance of our drug-injected rat 
subjects when they were knee-high.  I did have fun taking slides in to show 
their junior high school biology classes, when they learned microscopy, 
though it seems this was more exciting for me than it was for my kids (their 
classmates did send warmly appreciative notes thanking me!).

nostalgically,   Susan
- Original Message - 
From: "Breeden, Sara" 

To: "histonet" 
Sent: Thursday, March 11, 2010 8:09 AM
Subject: [Histonet] Histo Stories



Thanks to everyone that sent their Story of How I Ended Up Doing This
Histology Thing!  I have gotten 50 or more replies!  The one thing that
strikes me is how many of us went into this profession without a clue!
With all the opportunities to recruit future histologists, this
Histology Day idea is a good start. On the original subject, I'm
planning to make one document out of all the replies and - WITH
PERMISSION - attach your name to the answers.  If you do NOT want your
submission listed because you want to remain anonymous, you must let me
know ASAP.  Send to: nmhi...@comcast.net.  T

Re: [Histonet] Superfrost plus gold slides vs superfrost plus

[Susan Bachus]

Good old fashioned gelatin subbing is very cheap & works great--I have never 
seen tissue separate from a subbed slide!   Susan
- Original Message - 
From: "Peggy Bisher" 
To: "Milne, Katy" ; 


Sent: Friday, June 19, 2009 3:34 PM
Subject: Re: [Histonet] Superfrost plus gold slides vs superfrost plus


I had a group here who were having trouble with their cryo-sections 
sticking

to the slides. Their samples were rat brains. He found that the Gold Super
Frost Plus worked well with in-situ but not for immuno. At least that is 
the
comment he wrote down and put on our bulletin board in the lab. Most of 
the

people in the lab tend to use the Super Frost Plus (not the Gold) and they
seem pretty happy with them.


Margaret E. Bisher
Electron Microscopy & Histology Core Facility Manager
Department of Molecular Biology
Princeton University
Moffett Laboratory, Room 113
Princeton, New Jersey
Office: (609) 258-7026
Fax: (609) 258-8468
mbis...@princeton.edu





On 6/19/09 4:26 PM, "Milne, Katy"  wrote:


Hi everyone,

I'm curious if anyone has tried using Superfrost plus gold slides, are
they any better than the regular superfrost plus?  I'm dealing with
frozen lymph node on a Ventana discovery, I've tried a few different
fixing/drying combinations but I still have tissue lifting (not
completely but enough to be bothersome!).  Wondering if it's worth
giving the gold slides a go.

Thanks,
Katy

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Re: [Histonet] RNA In Situ Hybridization

[Susan Bachus]

Protocol #3 at this site is great for RNA probes: 
http://intramural.nimh.nih.gov/lcmr/snge/Protocols/ISHH/ISHH.html
This protocol assumes fresh frozen tissue but can be used on fixed, paraffin 
embedded tissue--obviously you would have to include a step at the beginning 
to deparaffinize.   You might also want to look at HIER methods that can 
improve results from fixed tissue.
- Original Message - 
From: "Bell, Pat" 

To: 
Sent: Wednesday, June 17, 2009 12:35 PM
Subject: [Histonet] RNA In Situ Hybridization


I want to learn to do In Situ Hybridization using RNA probes in formalin 
fixed, paraffin embedded tissues. I have the Dako Autostainer, but I know 
that some of the procedure would be done manually.


I would appreciate any protocols that would help me.

Thank you for your help.
Pat

Pat Bell HT(ASCP)
Medical Oncology; MS 8117
12801 E 17th Ave.
Aurora, Co. 80045
303-724-6077
pat.b...@ucdenver.edu


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Re: [Histonet] ISH probes

[Susan Bachus]

There's a wonderful company called Oligo's, Etc. (Wilsonville, OR) that 
synthesizes very affordable oligoprobes.  But you have to supply the 
sequence information.   Susan
- Original Message - 
From: "Sharon Campbell" 

To: 
Sent: Monday, January 26, 2009 9:20 AM
Subject: [Histonet] ISH probes


Hello Histonetters!

We are finding ourselves coming short on the time remaining that we can
offer ISH Probes through Ventana. We have not heard when or if ISH
(In-situ hybridization) will be offered any more through them. Does
anyone out there know of other companies that would offer ISH?

Thank you for your input.



Sharon Campbell, HTL(ASCP)CM, BSBM

Histology Supervisor

Celligent Diagnostics, LLC

Formerly Pathology Associates Services

101 East W.T. Harris Blvd, Suite 1212

Charlotte, NC  28262

800-524-6779 ext. 104

704-970-3304 Direct Line

shar...@celligent.net



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Re: [Histonet] subbing slides

[Susan Bachus]

Well, the attachment seems to have been deleted from what I sent, so here it 
is pasted below:
Gelatin-Subbed slides WEAR GLOVES! (not to protect your hands, but to 
protect the slides from RNase from your skin)


 a.. Place slides in racks and soak in hot tap water and soap (e.g. 
dishwashing soap, or Alconox) for at least 1 hr, but not more than a few 
hours (as soap could leave residue).

 b.. Rinse thoroughly in hot tap water until suds are completely gone.
 c.. Let sit in 80% ethanol for at least one hr. This solution can be 
re-used until it turns cloudy. Slides can be left overnight in this 
solution.

 d.. Rinse in a few changes of deionized water.
 e.. Subbing solution: Dissolve 3.0 g of gelatin (300 bloom swine) in 150 
ml deionized H2O by stirring while heating to 50 degrees C. Add 450 ml 
deionized water to cool, and then dissolve 0.3 g CrK(SO4)2*12H2O ("chrom 
alum") in the solution.
 f.. Filter this solution through Whatman Type I filter paper to remove any 
air bubbles. This solution is only good for the few hrs it will take for 
this batch of slides, as it will quickly solidify.
 g.. Dip the slides into the subbing solution, drain the slides onto a 
paper towel and allow to air dry for 1 hr.
 h.. Dip the slides into the subbing solution again. Drain and cover 
loosely with bench paper and allow to dry for a few hours.
 i.. When thoroughly dry, heat in oven at ~ 50 degrees C for a few hours, 
then store the slides in slide boxes in zip-lock bags. As long as they are 
kept dry, they should be good for years.
- Original Message - 
From: "Susan Bachus" 
To: "Neil Fournier" ; 


Sent: Tuesday, December 30, 2008 6:31 PM
Subject: Re: [Histonet] subbing slides



This always works like a charm for us and costs ~10x less than buying
pre-subbed or electrostatic or whatever fancy expensive store-bought 
slides!

Have fun! Susan
- Original Message - 
From: "Neil Fournier" 

To: 
Sent: Tuesday, December 30, 2008 6:21 PM
Subject: [Histonet] subbing slides



Hi Everyone,

Could someone share with me their procedure for cleaning and subbing 
glass

slides with gelantin? I am trying to work out a concentration of gelantin
and chromium potassium sulphate that will be optimal for adhering fairly
thick rat brain sections (30 to 120 um range depending on applications)

Much appreciated

Neil



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Re: [Histonet] subbing slides

[Susan Bachus]

This always works like a charm for us and costs ~10x less than buying 
pre-subbed or electrostatic or whatever fancy expensive store-bought slides! 
Have fun! Susan
- Original Message - 
From: "Neil Fournier" 

To: 
Sent: Tuesday, December 30, 2008 6:21 PM
Subject: [Histonet] subbing slides



Hi Everyone,

Could someone share with me their procedure for cleaning and subbing glass 
slides with gelantin? I am trying to work out a concentration of gelantin 
and chromium potassium sulphate that will be optimal for adhering fairly 
thick rat brain sections (30 to 120 um range depending on applications)


Much appreciated

Neil



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Re: [Histonet] Silly Question? - Need help quickly!

[Susan Bachus]

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 I am 
trying again with a URL for this paper:

http://www.jneurosci.org/cgi/reprint/13/4/1337

Susan

- Original Message - 
From: "Rene J Buesa" 
To: "Pat Flannery" ; ; 
"Weems, Joyce" 

Sent: Thursday, December 11, 2008 12:58 PM
Subject: RE: [Histonet] Silly Question? - Need help quickly!


Joyce:
Methanal, which is the chemical name of formaldehyde, polymerizes. If it 
forms a polymer of at least 50 molecules or more, it gets solid = 
para-formaldehyde.
Formalin (a trade name as formol is also another trade name)is the 37-50% 
aqueous solution of formaldehyde (with some additiveses to prevent 
polymerization).
You can prepare BNF using the formalin solution or dissolving the amount of 
solid para-formaldehydede to get to the concentrationon you desire.
The chemical in both solutions is the same = methanal or formaldehyde.René 
J.


--- On Thu, 12/11/08, Weems, Joyce  wrote:


From: Weems, Joyce 
Subject: RE: [Histonet] Silly Question? - Need help quickly!
To: "Pat Flannery" , histonet@lists.utsouthwestern.edu
Date: Thursday, December 11, 2008, 12:12 PM

I was just going to post a question regarding paraformaldhyde myself!
Just last week I believe I remember someone saying that paraformaldehyde
and formalin are the same and they had put the same solution in two
different containers for one of their researchers because they were so
insistent to have two different solutions. Are they the same?

Well, today I have a request to put tissue for a researcher in formalin
and paraformaldehyde. So Without percentage required, do I use 10%
NBF? Do I call somewhere and get paraformaldehyde and make 4%
paraformaldehyde?

I have asked the surgeon twice for the number for the lab so I can find
out - don't have it yet. I have two fresh adrenals in the fridge. Help!!


Thanks in advance...
Joyce

Joyce Weems
Pathology Manager
Saint Joseph's Hospital
5665 Peachtree Dunwoody Rd NE
Atlanta, GA 30342
678-843-7376 - Phone
678-843-7831 - Fax



-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pat
Flannery
Sent: Thursday, December 11, 2008 11:59 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Silly Question?

Please humor me on this if it's obvious (to everyone but me):  why do we
use paraformaldehyde (which is so inconvenient to make up) rather than
buffered formalin or just diluted formaldehyde itself?

It seems that around here, some folks prefer paraformaldehyde (either 2%
or 4%) and others use formalin, while some others stick to diluted
formaldehyde (I see all 4 on labels for specimens submitted for
histology).  Is it mostly a matter of personal preference or where you
were trained (i.e. force of habit) or is there a valid reason to use
each solution (basically the same chemical once in solution, merely
buffered or not)?  The only answer I've gotten when I've asked is,
"That's what we always use."

Thanks.

-Pat Flannery (not a "real" histologist - I just play one in the lab)


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Re: [Histonet] re: concentration of formaldehyde in soultion: ASTM D2194 - 02(2007) Standard Test Method for Concentration of Formaldehyde Solution

[Susan Bachus]

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
- Original Message - 
From: "tf" 
To: "Walters, Katherine S" ; "Tony Henwood" 
; "Pat Flannery" ; 
"histo...@lists.utsouthwestern.ed" 

Sent: Friday, December 12, 2008 10:12 AM
Subject: [Histonet] re: concentration of formaldehyde in soultion: ASTM 
D2194 - 02(2007) Standard Test Method for Concentration of Formaldehyde 
Solution




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...@lists.utsouthwestern.ed

抄送：
主题： RE: [Histonet] (reply) silly questions.---PFA

This may be another silly question, but how does one test the 
concentration of formaldehyde in solution?

Thanks,
Kathy
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-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu 
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of tf

Sent: Friday, December 12, 2008 2:35 AM
To: Tony Henwood; Pat Flannery; histo...@lists.utsouthwestern.ed
Subject: [Histonet] (reply) silly questions.---PFA
"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 less than 0.5%!!"
Tony: Do you think this is because of inproper preparation of PFA in his 
lab, or the common problem in all researchers using PFA?
I do think most biomedical labs currently are using PFA to prepare 
the fixatives!


So, anyone has the idea on a correction preparation procedure of 4% PFA?
I noticed some of you dissolve PFA powder in NaOH-conditioned alkaline 
water, then add concentrated PB solution.
We here dissolve PFA in concentrated PB solution directly (heat & stir for 
2-3 hours), then adjust pH to 7.4.
We dont have big problem in tissue quailityexcept when one want to cut 
the brain in a cryostat rather sliding microtome.
Many times the brain sections from the cryostat have "cheese" like 
holes/cavities, which almost never appear on sliding microtome-prepared 
sections.

2008-12-12
tf
发件人： Tony Henwood
发送时间： 2008-12-12  06:18:47
收件人： Pat Flannery; histonet@lists.utsouthwestern.edu
抄送：
主题： RE: [Histonet] Silly Question?

Pat,
I agree with you.
In a routine diagnostic histopathology laboratory, it makes little
difference what you use. Around the world for over 100 years most labs
use 10% neutral buffered formalin made from concentrated 38%(or there
abouts) formalin (or formaldehyde).
Researchers, though, are a different kettle of fish. They will tend to
hang on to misinformed, "mystical" methods believing they are being
scientific. Funny, you would think that they, as a group, would be the
ones pushing the boundaries and critically assessing each step of their
research, ensuring that they understand what and why they are doing it.
(Disclaimer - not all researchers are like this, thank heavens!!)
Using a formaldehyde solution made from polyformaldehyde can cause
problems. One researcher used it and wondered why their morphology was
sub-optimal and their p53 immunohistochemistry was negative. He assured
me that he had fixed small samples of tissue for 6 hours in 4%
formaldehyde and then processed them using ethanol, xylene and wax.
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 less than 0.5%!!
This also explains the loss of p53 staining. I gave him some of our
routine 10% phosphate buffered fomalin, asked him to fix overnight, and
try agin. Low and behold problem solved.
How's that for a Friday Flamming!!!
Regards
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

Re: [Histonet] Alias identity

[Susan Bachus]

Here's one potential problem with listing real names:  a few years ago a 
very diligent student of mine inadvertently stirred up a lot of controversy 
when she asked for advice from Histonet on a class assigment and someone was 
concerned that she was "cheating".   I had in fact recommended Histonet as a 
resource to them (though I didn't intend for them to use it for something so 
trivial)!   One silver lining was that a lot of discussion was aired about 
what the purpose of Histonet is and several people sprang to the poor 
student's defense.  But later she googled herself when she started worrying 
about looking for a job and this popped up because she had used her full 
name, and she was terrified that it would interfere with her finding a job! 
She wanted to know if it could be taken out of the archives, but of course 
the problem is that these things are still "cached"!   Susan
- Original Message - 
From: "Charles.Embrey" <[EMAIL PROTECTED]>
To: "Amber McKenzie" <[EMAIL PROTECTED]>; 


Sent: Wednesday, December 03, 2008 1:30 PM
Subject: RE: [Histonet] Alias identity


This is one problem you have with an open list.  I also belong to the
Pathologists' Assistants list server but you must apply for membership
that is restricted to AAPA members only.  We can be more open with
comments and questions without worry of unwanted calls from vendors or
junk mail from the growing number of recruiters.  Of course anything on
the web can be accessed if you search hard enough but we don't have
companies monitoring everything we say.  I would be careful to "assume
that if we're on this list that we're all credible, knowledge and
professionals".  Over ten years on the list has taught me otherwise.  I
must admit that I always consider the source.  It is sad that some do
have to hide their identity but I do understand why they find it
necessary and support their choice.

Chuck

-Original Message-
From: [EMAIL PROTECTED]
[mailto:[EMAIL PROTECTED] On Behalf Of Amber
McKenzie
Sent: Wednesday, December 03, 2008 10:06 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Alias identity



I have to put my 2 cents in about all the statements on having an alias
name instead of identifying who each person is after each email...WHAT'S
THE BIG DEAL??  We are all supposed to be professionals asking each
other for advice/suggestions on the Histonet - who cares who each person
is?  If I post a question, I don't care if it's Jane Doe answering or
John Smith.  I assume that if we're on this list that we're all
credible, knowledge and professionals.  Come on people, we're all in the
same boat here. If I'd thought about putting an alias name for myself
instead of my real name, I would have! Simply b/c last month I posted a
question on the Histonet asking where you all bought your lab chairs,
and I ended up having 2 vendors call me at my office trying to sell me
some when all I wanted was the advice of other HT's.  I love the idea of
people not knowing who I am or where I work.  Then I wouldn't have to
worry about being harassed on the phone.



Instead of worrying about the little things on the Histonet - like who
each person is, why can't we focus on work related issues?  I have to
delete so much junk just to get to the material that actually applies to
my field b/c of all the multiple emails that don't even matter.

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Re: [Histonet] histology ink washing off

[Susan Bachus]

Back in the old days we used to use "india ink" to label slides--so I don't 
know whether it would also work on tissue (as opposed to glass), but it 
might be worth a try.   Susan
- Original Message - 
From: "Gudrun Lang" <[EMAIL PROTECTED]>

To: "'Leslie Chen'" <[EMAIL PROTECTED]>
Cc: 
Sent: Saturday, October 11, 2008 5:15 AM
Subject: AW: [Histonet] histology ink washing off


We dry and heat the tissue surface with a hair-dryer at the grossing. It
sounds a little bit rigid, but it works and the tissue margins are easily
seen in the slide. The excess dye also will be solved in the NBF, but there
sticks enough on the tissuemargins.

Gudrun Lang

Biomed. Analytikerin
Histolabor
Akh Linz
Krankenhausstr. 9
4020 Linz
+43(0)732/7806-6754
-Ursprüngliche Nachricht-
Von: [EMAIL PROTECTED]
[mailto:[EMAIL PROTECTED] Im Auftrag von Leslie
Chen
Gesendet: Freitag, 10. Oktober 2008 22:31
An: histonet@lists.utsouthwestern.edu
Betreff: [Histonet] histology ink washing off

Hello,
I am having a problem with histology tissue marking ink washing off
specimens in water, PFA, or 70% alcohol.  This is before the tissue even
gets processed for paraffin embedding. Does anyone have any suggestions on
how to keep the ink from coming off?  Thank you.

Leslie


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