[Histonet] Pathology Ladder System

[Barry Rittman via Histonet]

A well defined career ladder has often been a missing factor in salaries
for histotechs.

I believe that part of this is due to the change in the Med Lab Tech
training System.
Originally this system used to incoporate histology in the training.
I believe in the early 70s the histology section was removed from the
training and histopatholgy was on its own.
This, in  effect resulted in Med lab techs getting a degree and often being
placed in charge of labs including histology/pathology labs without realy
having much if any experience in these areas.
I trained in England where histology was one of the six areas in med lab
tech training in the entire country. Following this training it was
possible to specialise in one of the fields such as histopathology.
This provided  a direct career path as everyone working in a lab all had
the basic training and knew what to do to advance in their career.
During my career, while I worked in histology (and this is still my first
love), I was able to draw on my training in the other fields to help with
my histopathology.

I think that in effect the system of often having med lab techs in charge
of histopatholgy have artifically kept salaries for histopathogists
artificially low.

I personally would prefer the med lab tech training (including histology)
with a direct career path for all but am not holding my breath that this
will occur
Barry.
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Re: [Histonet] Air Bubbles

[Barry Rittman via Histonet]

There might be air in the mounting media that gives rise to bubbles when
drying starts to occur. One method to try is to use an ultrasonic bath  to
remove the air from the bottle containing the mounting media  before using.
We used to do this with aqueous mounting media.
Barry

On Fri, Jun 10, 2016 at 9:15 AM, Karen Pfaff via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

>
>
>
> There might not be enough mounting media placed on slide. So when slide is
> wet, there are no air bubble but after a couple of days the xylene or
> xylene substitute will evaporate and leave you with patchy areas. Has there
> been someone new coverslipping? I have been one of those people that put
> less mounting media on because the stage of the microscope stays cleaner,
> however, I discovered that I get air bubbles after it dries. Try putting a
> line of mounting media down the entire slide not just 2 or 3 drops. Might
> help.Karen PfaffLead HT Skin Cancer CenterMohs lab
>
> Sent from my U.S. Cellular® Smartphone
>
>  Original message 
> From: Charles Riley via Histonet 
> Date: 06/10/2016  9:00 AM  (GMT-06:00)
> To: histonet@lists.utsouthwestern.edu
> Subject: [Histonet] Air Bubbles
>
> Hello everyone,
>
> Recently we have been having an issue with a majority of our slides having
> air bubbles under the coverslips. When they are originally coverslipped
> there are no bubbles and any present are pushed out and the sides are all
> wiped dry. We have not changed any reagent or processes in over two years.
> Can anyone offer suggestions as to why this might be occurring or tips to
> fix the issue?
>
> --
>
> Charles Riley HT(ASCP)CM
>
> Histopathology Coordinator/ Mohs
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Re: [Histonet] Clearance Angle on Microtome Blade

[Barry Rittman via Histonet]

Rene thank you.
There is an excellent book on the section cutting that will give you any
theory you need.
Section Cutting in Microscopy. by H.F. Steedman. 1960. Blackwell Scientific
Publications. Oxford.
Also an American Optical Booklet on the Effective Use and Proper Care of
the Microtome.
Barry

On Sat, Jan 30, 2016 at 7:37 AM, Rene J Buesa via Histonet <
histonet@lists.utsouthwestern.edu> wrote:

> The type of tissue, the speed of sectioning, the knife bevel and the type
> of paraffin (melting point) influence the clearance angle.Anywhere from 5
> to 10º (preferable 5-6º) are the most used.René
>
> On Friday, January 29, 2016 3:17 PM, Kelli Goodkowsky via Histonet <
> histonet@lists.utsouthwestern.edu> wrote:
>
>
>  Good afternoon,
> Can someone touch base with me regarding the clearance angle of the
> microtome blade?  How would you describe a blade clearance angle that is
> too great or too slight, and what microtomy problems are you likely to see
> with each?  There seems to be conflicting information out there (or at
> least varying perspectives).
>
> Thank you!
> Kelli
>
> Kelli Goodkowsky, M.Ed., HT (ASCP)
> Director Clinical Education
> Histologic Science
> Goodwin College
> (860) 727-6917
> kgoodkow...@goodwin.edu
> http://www.goodwin.edu/
>
>
>
>
>
>
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>
>
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Re: [Histonet] are you desperate to hire a BS.c graduate

[Barry Rittman via Histonet]

Jose brings up some good points.
It seems to me that we often spend too much time  talking about employing
people who have a narrow field of expertise, even though such expertise is
necessary to get the job done. I would suggest that we instead look at the
bigger picture.
Experience  outside your current field of work  can be very useful. In
looking at the histology fields, many of the great advances have not always
been made by people who have several years or decades of experience but
often by individual who had very limited histology experience but had
expertise in other fields such as biochemistry and physics. I do not
believe that any experience no matter how far removed from the current
workplace is wasted.  While a bachelors degree may not prepare individuals
adequately for jumping right into the field of histology, it has at the
very least  given them a different outlook on many things that they would
not have without that degree.  Fresh outlook on our work allows us to
improve the work we are doing and be creative instead of automatons. I
think that the field of histotechnology is always improved by having
individuals of different back grounds bringing  different perspectives.
The most important thing is to let individuals explore  any new ideas they
have.
I would say a lot more but my wife is just serving up a beef pie.
Barry

On Thu, Aug 13, 2015 at 2:47 PM, deGuzman, Jose R via Histonet 
histonet@lists.utsouthwestern.edu wrote:

 Please correct me if I'm wrong but this has been my experience so far.

 I came from a research Histology background where I learned many skills
 that I was able to developed in the clinical side. I spent 3 years
 preparing for the exam because my manager in research and a co-workers
 failed on their first attempt- they scared me. Now, I supervise the same
 clinical lab that took a chance at me and perform the hiring process. I
 have hired both certified and
 not-yet-certified-but-qualified-to-take-the-BOR-exam-for-certification-within-1-year.

 Histology is a field where you will find a wide range of experience and
 skill sets. You will find individuals with years of experience with a
 narrow skill set and an individual with little months/years of experience
 but have a wide range of skill set and troubleshooting experience to boot.
 These variations are the direct result of what the laboratory exposed us
 to- large, high volume versus small volume, specialized labs. If you find
 someone with years of experience and wide range of skills, they probably
 work with you already because we don't let them go.

 As a supervisor, I look at the following: qualifications (must meet the
 minimum requirements to sit for the exam), experience, skill sets, ability
 to learn and adapt and fit. The first, will eliminate anybody who I cannot
 hire due to our hiring policy. Once you meet that first criteria,
 everything else is a sliding scale. Fit will out-weigh experience because
 you can build experience but if you can't get along with my team, there is
 no team. I will work with anybody to develop their skills as long as they
 are able to learn and adapt. I'm in an area where there's strong
 competition for qualified individuals and I know of only 1 school with a
 Histology program that's over 50 miles away. It's not easy to fill
 positions. So for the research Histologists out there, yes it's very
 different. It's very routine, not much if any variation from day to day,
 month to month. We have to crank the cases out and some of us get
 pigeonholed to just embedding or just cutting for 8-12 hours. Can we make
 the transition? Yes, I know because I did it.

 Different organizations have different hiring policies. Full package
 candidates (certified, experienced, skilled) are rare but it makes the
 on-boarding and the first 90 days so much easier. Especially in a very busy
 lab, bringing someone in who can contribute at the same output as the
 established team is a dream come true. Incomplete packages need time to
 develop. And what I've experienced is research histologists may know a lot
 more than I do and are exposed to other aspects of the lab but when it
 comes to getting the work done, there is a lag that requires me to step-in
 because the work is taking longer to complete. The learning curve is steep,
 it doesn't take long. It's just easier to bring in a complete package.

 Jose



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[Histonet] measuring cells

[Barry Rittman]

I apologize for misplacing the original posting about this.

 Measuring the number of cells in sections is very tricky and varies
considerably with the thickness of the section and the size of the cells.
In general the thicker the section the more accurate the count as there are
more complete cells rather than just profiles. The problem is that in order
to get a really accurate count you need sections that are so thick that
resolution suffers. Abercrombie in 1946 published papers that took into
account the size of cells and the thickness of the sections and depending
on these applied a correction factor that improved  the accuracy of the
count. His papers are worth reading. The link is below for those who are
interested.


http://www.nervenet.org/papers/Abercrombie46.html

Of course an interesting approach would be to cut several sections,
separate the cells and then use a Coulter counter... like this is going to
happen!!

Barry
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Re: [Histonet] Stain vs dye and control

[Barry Rittman]

There is always confusion about this.
In textile coloring the term dyeing is used and this first carried over
to histology as many of the textile dyes were the ones first used to stain
tissue components. Some of these were specific and some non specific.

In histology my understanding is that the dye is the powder, while the
stain is the solution used for staining the tissues. and in most cases
there is a certain amount of specificity in the reaction. Merely imparting
a non specific color all components of a tissue I would regard as coloring
as there is no specificity in this process.

This raised the interesting question of what would you call the process
whereby osmium tetroxide is used to fix and stain some fats?

To further complicate in the concrete industry the term dye is used to
indicate a general coloring of the concrete while the term stain  is used
to indicate a chemical reaction with the concrete components.
Barry


On Sun, Mar 8, 2015 at 3:02 AM, Gudrun Lang gu.l...@gmx.at wrote:

 Hi Jorge,
 Not all histolabs have access to fresh cultures. So they search an easy
 way to get bacterial controls. And things like sausage are more like the
 usual specimens than liquid samples.

 The best staining control is an inhouse-specimen, that is processed in the
 same way as any other specimen (preanalytic, fixation, processing, cutting,
 staining). But sometimes this is not possible, so one uses the next-best.
 A control with known ingredients (like bacteria) can be used to check the
 whole process and must be positive for the tested parameter. A
 patient-sample can only be considered positive or negative, if the
 positive-control proves the functionality of the process
 (staining-protocol).

 Dye and stain. You can touch the dye, but not the stain.  And then you
 have got stained fingers. ;-)

 Gudrun



 -Ursprüngliche Nachricht-
 Von: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Jorge A.
 Santiago-Blay
 Gesendet: Freitag, 06. März 2015 21:49
 An: Histonet@lists.utsouthwestern.edu
 Betreff: [Histonet] Stain vs dye and control

 Dear Histonetters:

 Last semester I taught a microbiology lab and, as I was reviewing for
 class, noticed some lack of precision in the use of the terms dye vs.
 stain in biology. Could someone help?

 While I am on stains, I have been following the emails on controls and
 wonder a couple of things.

 1. What would testing for bacteria on beef jerky, hot dogs or burgers
 accomplish that is different (ideally better) that what one accomplishes by
 pulling out from fresh cultures out of a medium (e.g. liquid, such as
 broth, solid, such as slabs, agar)?  Is it the idea to test for bacteria in
 an animal tissue? If so, would a solid medium (like someone mentioned
 recently, such as agar) do?

 2. An advantage of using fresh bacterial cultures of known Gram is that it
 could be used to test whether the reagents are good enough. Last semester I
 had the suspicion that one (or more) of our Gram reagents where not up to
 par.

 If you have any feedback, please feel free to email me directly at
 blayjo...@gmail.com . Thank you.

 Sincerely,

 Jorge

 Jorge A. Santiago-Blay, PhD
 blaypublishers.com
 http://blayjorge.wordpress.com/
 http://paleobiology.si.edu/staff/individuals/santiagoblay.cfm
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[Histonet] honey versus formalin

[Barry Rittman]

Sorry but my itchy delete finger got rid of the subscriber who was asking
about this.
There is a 2014 article by Sabarinath et al in Journal of Histotech. vol 37
pp 21-25.  comparing honey and formalin fixation.
If you use honey versus formalin as fixative and go to the go to
www.maneyonline.com  refernce you can access the entire article.
Barry
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Re: [Histonet] processing problem

[Barry Rittman]

Hi Martha
I think that Caroline is correct but you have nothing to lose by removing
the  wax and then trying to dehydrate and so on. A more gentle way after
removing the wax and dehydrating is to use chloroform instead of xylene and
hand processing. Chloroform is much more gentle although tissue do usually
float in it an do not become transparent. However unlike xylene, tissue can
be left in chloroform for hours to overnight without any appreciable
hardening.  Then after chloroform you can leave the tissues in a mixture of
chloroform/wax (1:1) at room temperature again for hours to even overnight.
During this process some wax does penetrate into the  tissue and therefore
can leave in wax for less time with less chance of hardening.
Yours is certainly not the first lab that this has happened to.
Good luck to you.
Barry

On Mon, Feb 16, 2015 at 11:30 AM, Caroline Miller mi...@3scan.com wrote:

 I am afraid your tissue might be 'toast'. I tried to reprocess from that
 same issue and things did not go well. We were processing mouse tissue,
 which is dry enough, but then when we put the tissues back through xylene
 and into 100% and back again it got even worse.

 We had to just get what we could from the blocks and confess to the
 researchers what had happened :(

 sorry for the bad news, I hope someone has better for you!

 mills

 On Thu, Feb 12, 2015 at 5:41 AM, Davenport, Martha R 
 martha.davenp...@uky.edu wrote:

  I am Martha Davenport, supervisor, at the University of KY histology lab.
  We had a processing problem caused  by  accidently placing 70% (instead
 of
  100%) into the last dehydration container. Would anyone please give me
 info
  on how you would remedy this?  We usually reprocess the tissue but have
 had
  trouble in the past with the tissue morphology being optimal. Any help
 will
  be greatly appreciated.
  Martha Davenport 859-257-1822
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 --
 Caroline Miller
 Director of Histology
 3Scan.com
 415 2187297
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Re: [Histonet] Re: Reference to microtome micrometer thickness

[Barry Rittman]

This has to do with section thickness. Would ne nice is did no0t have
several other topics un der the same heading.

Hi had a senior moment earlier so now will try to complete my sentences
before pressing the send key..
I am not sure that it is essential to know accurately the thickness of
sections, but the consistency of the sections in a ribbon.
The microtome setting is only an approximate guide as the  section
thickness depends on many factors including the tissue density and
homogeneity, temperature of the block and room, speed and angle of cutting,
time between individual sections, whether the block has been moistened with
ice water between sections and finally the skill and mood of the operator.
In limits between 10 and 1 microns the thinner the sections the greater the
chance of uneven thickness between sections. I think that Steedman's book
on section cutting explained that an individual section is cleaved from the
block, and like cheese slicing  with a regular knife (sorry I know that you
are all recovering from food overload) the thinner the slice the tendency
is for less consistency between individual slices.
If you cut a ribbon of sections then the consistency of individual sections
once the slide is stained should be fairly obvious and easy to measure
using image analysis of density and/or color. It is possible with
experience to
determine the difference between 4 and 7 micron sections but I do not
believe the difference of 1 micron between individual sections can be
easily determined.

There are a couple of ways to be very accurate but I am not sure the effort
is really worth it.
1.  Use a homogenous block of protein such that is radiolabeled,  embed and
section with your block of tissue and use a counter on sections to
determine how much activity and therefore thickness of individual sections.
2.  Use a block of protein stained with solvent resistant dye and measure
using image analysis. As you know the concentration of the dye you can
determine how much is in an individual section. Many Procion dyes (chloro s
triazines)  bind tenaciously to proteins and are resistant to most chemical
used in the histology lab.
3. Use labeled or stained micro-spheres of known diameter in the block and
determine how many profiles you have when focusing through a section.
4. Use built in markers such as red blood cell in small vessels, there
should be enough there to determine approximate thickness.

While this an interesting problem I would suggest that the easiest
approach, unless you have no social life,  is to rely on the skill of the
microtomist.
Barry


On Fri, Nov 28, 2014 at 3:47 AM, Vernon, Richard I. 
richard.ver...@thermofisher.com wrote:

 Hi Maxim,

 Does measuring section thickness on slide scanners get affected by the
 surface imperfections on the section?

 It has been indicated to me that surface imperfections certainly hinder
 scanning speed but what about the actual thickness measurements?


 Kind regards

 Richard Vernon
 Strategic Product Manager - Sectioning Products
 Anatomical Pathology Division

 Thermo Fisher Scientific
 Tudor Road, Manor Park, Runcorn, Cheshire, WA7 1TA, UK
 Tel:+44 (0) 1928 534122 | Mobile:+44 (0) 7825 119070
 richard.ver...@thermofisher.com | http//:www.thermoscientific.com





 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
 histonet-requ...@lists.utsouthwestern.edu
 Sent: 28 November 2014 09:43
 To: histonet@lists.utsouthwestern.edu
 Subject: Histonet Digest, Vol 132, Issue 29

 Send Histonet mailing list submissions to
 histonet@lists.utsouthwestern.edu

 To subscribe or unsubscribe via the World Wide Web, visit
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 Re: Contents of Histonet digest...


 Today's Topics:

1. Re: Reference to microtome micrometer thickness (Barry Rittman)
2. Congo Red problems (Wheelock, Timothy R.)
3. Re: Congo Red problems (Carlos Defeo)
4. RE: Formalin smell in the last paraffin station in vip6
   tissue tek (Jamal)


 --

 Message: 1
 Date: Wed, 26 Nov 2014 12:05:53 -0600
 From: Barry Rittman barryritt...@gmail.com
 Subject: Re: [Histonet] Reference to microtome micrometer thickness
 Cc: histonet@lists.utsouthwestern.edu
 histonet@lists.utsouthwestern.edu
 Message-ID:
 
 ca+0tsf3q_yxa2y2bjrgxef8tix_faw6miv-bnayme75x7-h...@mail.gmail.com
 Content-Type: text/plain; charset=UTF-8

 It has been a while since I read articles regarding section thickness but
 I have some comments:
 1.

 On Sat, Nov 22, 2014 at 8:37 PM, Maxim Peshkov

Re: [Histonet] Reference to microtome micrometer thickness

[Barry Rittman]

It has been a while since I read articles regarding section thickness but I
have some comments:
1.

On Sat, Nov 22, 2014 at 8:37 PM, Maxim Peshkov maxim...@mail.ru wrote:

 Reuel,
 It is right, that there are many external reasons for a thickness of
 paraffin sections.
 It is possible to measure the final stained sections with mathematical
 methods by slide scanner.
 I hope that the slide-scanner can be easily to calculate thickness of
 section by their soft.
 Each individually sections will have own specific thickness.

 Sincerely,
 Maxim Peshkov,
 Russia,
 Taganrog.
mailto:maxim...@mail.ru


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Re: [Histonet] Phloxine-Tartrazine Stain

[Barry Rittman]

Wendy
if I am not mistaken, and understand that it is early in the day,  phloxine
absorbs at 548nm and tartrazine at around 425nm so both can show
fluorescence depending on your filtering system.

Barry

On Mon, Oct 6, 2014 at 1:49 PM, Trunch, Wendy L wl...@tulane.edu wrote:

 Hi All,

 I am trying to stain using Phloxine-Tartrazine and then follow with
 immunofluorescent staining. Has anyone tried this? Any suggestions?

 Thanks,
 Wendy
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Re: [Histonet] RE:On the lighter side...

[Barry Rittman]

Well fixed in the histological sense I hope.
Barry


On Mon, Aug 11, 2014 at 2:52 PM, Mayer,Toysha N tnma...@mdanderson.org
wrote:

 Tim,

 Just saw your post about the 'marketable skill'.  Funny those were my
 mother's exact words while I was in college.  She didn't care what I
 majored in, as long as I got a marketable skill along the way.
 The best advice I've ever gotten.
 Thanks Maria!!! (my mother)

 Sincerely,

 Dr.Toysha N. Mayer, D.H.Sc., MBA, HT (ASCP)
 Instructor/Education Coordinator
 Program in Histotechnology
 School of Health Professions
 UT M.D. Anderson Cancer Center
 713.563-3481



 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy
 Sent: 08 August 2014 19:26
 To: 'Douglas Porter'; histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] On the lighter side...

 Wow, I feel like a newbie! 28 years, registered. HT 13663, 1988, HTL 1369,
 1992. Electron Microscopy Technologist, #604, 1982.

 Like most, I never heard of histology until I walked into a hospital lab
 on my first day as  an EM tech. I had seen slides made in college, but no
 one ever mentioned it could be an actual profession. I was more taken with
 the electron microscope, and there was (is) a 2-year program at the
 community college in the town I grew up in (Delta College, Stockton, CA).
 So AFTER getting a BA in Zoology, I went there to get a marketable skill.
 At that time EM was still used for tumor dx, so when I started it was about
 half tumor, half kidney. I was lucky enough to get involved in histology
 and set up the IHC lab at the small community hospital I worked at (as an
 EM tech) and so ended up phasing myself almost out of an EM job. The IHC
 took over all the tumor dx from EM. Later I left EM altogether and did IHC
 exclusively for 15 years. But, like most, I learned Histotechnology on the
 job but was lucky enough to work for a pathologist who believed in
 developing his techs - to the point of paying for meetings out of his own
 pocket. Only now do I know how fortunate I was to work for someone like
 that. Because of him we had developed a culture in the small histo lab (4
 men!!) of learning. We studied together one night a week for the HT exam
 and all passed (and the practical!). Again, that was a fortunate
 experience, not very often seen in labs.

 Tim Morken
 Supervisor, Histology, Electron Microscopy and Neuromuscular Special
 Studies UC San Francisco Medical Center San Francisco, CA



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Re: [Histonet] Sources of LVN (low viscosity nitrocellulose)

[Barry Rittman]

Technically LVN and celloidin are not quite the same.
LVN (also known as gun cotton) is useful for large specimens -we used to
use this for aborted fetal heads and large bone specimens. It penetrates
much more rapidly that celloidin and is much easier and safer to handle.
Tissues prepared especially in celloidin are magnificent due to the very
low rate of shrinkage.
The cost of safe transport of these is prohibitive.
I am retired now but in the 1990s celloidin from Fisher was $1,200  a pound
plus transport costs.
Barry

.


On Thu, Aug 7, 2014 at 3:01 PM, Willy Bemis we...@cornell.edu wrote:

 Historically, we used LVN (low viscosity nitrocellulose, also marketed as
 Celloidin and various other trade names) to embed specimens such as whole
 decalcified salmon heads for thick sectioning (30 to 100µm) using sliding
 microtomes such as the AO 860.

 I used to be able to purchase LVN powder wetted with alcohol from the
 Hercules Company. Because their LVN  was not highly purified, it was much
 less expensive than Celloidin. It worked perfectly for our technique.
 Unfortunately, Hercules is no longer in business.

 Does anyone know of a commercial source for LVN?

 Willy



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Re: [Histonet] Protocol for decalcifying and processing large bone section.

[Barry Rittman]

I would recommend using Kristensen's sodium formate/formic acid mixture.
This is more gentle than formic acid alone and almost as rapid.
Barry


On Tue, Aug 5, 2014 at 12:01 PM, DiCarlo, Margaret 
mdica...@kaleidahealth.org wrote:

 I use 10% formic acid for large human bones but I don't think your bone
 sections are that large.

 Peggy DiCarlo
 Ortho Bone Lab
 BGMC
 100 High St.
 Buffalo, NY  14203

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Clough, Bret
 Sent: Tuesday, August 05, 2014 11:55
 To: Histonet list serv.
 Subject: [Histonet] Protocol for decalcifying and processing large bone
 section.

 Hi everyone,
  I was hoping someone on histonet would being willing to help me by either
 sharing their protocol or advising me on decalcifying and processing large
 bone sections. The bone sections are from the femural head of sheep
 measuring roughly 1cm x 1 1/2 cm cube.  Currently I've been decalcifying
 the samples in 0.5M EDTA which is taking along time to decalcify this being
 the 19th day.  Any advise would be greatly appreciated!


 Sincerely,
Bret Clough
Texas AM Health Science Center
Temple, TX.
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Re: AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde

[Barry Rittman]

hi
I would recommend storage for long term in 70% ethanol. To prevent drying
out we used glycerin in the ethanol, about 20% of the volume.
Barry




On Mon, Dec 9, 2013 at 7:44 AM, Orla M Gallagher 
o.m.gallag...@sheffield.ac.uk wrote:

 Thanks to everyone for your comments.

 I may not have been clear in my question - our researchers don't wish to
 decalcify these formalin-fixed bones yet, but rather to store them for more
 than a couple of weeks, in case they need to carry out MicroCT followed by
 histology later. I'm aware that the formalin or paraformaldehyde will
 degrade over time, but I just wondered if anyone has a protocol for storage
 without decalcification? I guess transfer to 70% ethanol is an option but
 this is also not ideal for longterm storage, and would need to be removed
 before decal in EDTA.

 All the best,
 Orla


 On 6 December 2013 16:12, Wineman, Terra terra.wine...@novusint.com
 wrote:

  I would suggest a different protocol if the tissue will not be processed
  for a while.  I would say a week in 10%NBF and then transfer the bones to
  an EDTA decal solution.  The bones will decal slowly without the affects
 of
  the formic acid.  I am in research and this is what we do with our bones.
 
  Terra Wineman, HTL (ASCP)CM
  Research Biologist
  636-926-7476 phone
  terra.wine...@novusint.com
 
 
  -Original Message-
  From: histonet-boun...@lists.utsouthwestern.edu [mailto:
  histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
 pru...@ihctech.net
  Sent: Thursday, December 05, 2013 2:50 PM
  To: gu.l...@gmx.at; 'Orla M Gallagher'
  Cc: histonet@lists.utsouthwestern.edu
  Subject: RE: AW: [Histonet] Bone samples long-term storage in 10%
 formalin
  or 4% paraformaldehyde
 
  i would think u are correct in advising formic acid decal and then
  processing into paraffin for the best protection of the trap enzyme,
  immunoreactivity, etc.  A couple of weeks in formalin should be fine.
   Paraformaldehyde show be the same as formalin.  I do know a way to
 restore
  the enzyme activity for TRAP that may have been lost so if u need that
 let
  me know.
 
  - Original Message - Subject: AW: [Histonet] Bone samples
  long-term storage in 10% formalin or 4% paraformaldehyde
  From: Gudrun Lang gu.l...@gmx.at
  Date: 12/5/13 11:42 am
  To: 'Orla M Gallagher' o.m.gallag...@sheffield.ac.uk
  Cc: histonet@lists.utsouthwestern.edu
 
  Paraformaldehyd is formaldehyd in solid form. Formalin is the aequous
   solution of formaldehyd.
   So the main characteristics are the same.
 
   Gudrun Lang
 
   -Urspruuml;ngliche Nachricht-
   Von: histonet-boun...@lists.utsouthwestern.edu
   [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Orla
 M
   Gallagher
   Gesendet: Donnerstag, 05. Dezember 2013 19:31
   An: histonet@lists.utsouthwestern.edu
   Betreff: [Histonet] Bone samples long-term storage in 10% formalin or 4%
   paraformaldehyde
 
   Dear Histonetters,
 
   What is your opinion on storing bone samples long-term (more than a
  couple  of weeks) in 10% formalin? As I was taught, best practice has
  always been to  fix only as long as necessary, depending on the size of
 the
  sample, then  decalcify and process to wax, and I always stress this to
  everyone I advise.
 
   However, research colleagues sometimes wish to do histology on bone
  samples  that have been stored for months ..or even years! As the
 formalin
  pH becomes  more acidic, there is formalin pigment and the
 immunoreactivity
  and TRAP  enzyme activity is diminished or destroyed during long
 fixation,
  is there  any way of minimising this e.g. has anyone tried regularly
  replacing the old  formalin with fresh buffered formalin, or storing
  formalin-fixed bones in  any other medium? I'm also interested in how
 best
  to fix in 4%  paraformaldehyde and whether the problems are the same with
  long-term  storage.
 
   Thanks for your comments.
 
   All the best,
   Orla
 
   --
   **
   Ms. Orla Gallagher
   Bone Analysis Laboratory
   Mellanby Centre for Bone Research
   Department of Human Metabolism
   D Floor Medical School
   University of Sheffield
   Beech Hill Road
   Sheffield
   S10 2RX
   UK
 
   Website: http://mellanbycentre.dept.shef.ac.uk
 
   Tel: 0044114-2713337 (office)
   0044114-2713174 (lab)
   E-Mail: o.m.gallag...@sheffield.ac.uk
 
 
   *STOP*: Do you really need to print this e-mail?
 
   *BE GREEN:* Keep it on the screen.
 
 
   *Times Higher Education University of the Year*
 
 
 
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   The information contained in this message or any appended documents may
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Re: [Histonet] rat's hind paw skin

[Barry Rittman]

Hi Leila
As no one seems to have responded to your email I will add a few items.
I used to do some work on wound healing on mouse footpads so circumstances
are the same.
If you are looking for melanin routine fixation and processing should be
finem, melanin will remain.
Only thing is that the epidermis is tough and there is not much dermis in
this region.
Need to make sure that you are careful in cutting sections or the epidermis
will separate.
Hope this helps
if you need any more details please email me.
Barry



On Mon, Dec 2, 2013 at 3:44 AM, Leila Etemadi leila.etem...@med.lu.sewrote:

 Hello every body,

 First I would like to thank you all for your great feedbacks and attention
 which is such a big improvement in my work, Thank you all!

 Then, I was wonder if any one has experience on this tissue: rat hind paw
 skin ( which is hairless).

 Any inputs will be appreciated, whether is about any specific source where
 I can find technical information about fixation procedure..., or about
 sectioning,staining procedure…

  I am looking for melanin pigments and have no clue if it is possible to
 see it there or not?!?

 Thanks in advance,

 Cheers

 Leila




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Re: [Histonet] RE: Histo arcania ... Reticulum vs Reticulin

[Barry Rittman]

Reticulin is a type of collagenous fiber that is generally argyrophilic and
first recognized as a type of collagen with the advent of electron
microscopy. Associated with basement membranes and with fiber networks in
bone marrow, lymph nodes  etc.
Reticulum in* anatomy* refers to a network.
In* histology* it is usually used to describe the endoplasmic reticulum
organelles in cells, comprised or smooth and rough endoplasmic reticulum
and involved in protein synthesis and modification.
Barry


On Thu, Nov 7, 2013 at 5:48 PM, Weems, Joyce K. 
joyce.we...@emoryhealthcare.org wrote:

 I have seen them and have been told they are the same and interchangeable.
 I've never really looked into it, tho. It will be good to learn!

 Joyce Weems
 Pathology Manager
 678-843-7376 Phone
 678-843-7831 Fax
 joyce.we...@emoryhealthcare.org



 www.saintjosephsatlanta.org
 5665 Peachtree Dunwoody Road
 Atlanta, GA 30342

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 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Morken, Timothy
 Sent: Thursday, November 07, 2013 6:03 PM
 To: Histonet
 Subject: [Histonet] Histo arcania ... Reticulum vs Reticulin

 Oh Great Histonet, how do you describe the difference, if any, between the
 terms reticulum and reticulin.



 Tim Morken
 Supervisor, Electron Microscopy and Neuromuscular Special Studies UC San
 Francisco Medical Center Box 1656
 505 Parnassus Ave
 San Francisco, CA 94143
 USA

 415.353.1266  (office)
 tim.mor...@ucsfmedctr.orgmailto:tim.mor...@ucsfmedctr.org


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Re: [Histonet] Kisser's Mounting Media (Glycerol jelly)

[Barry Rittman]

You can use thymol or merthiolate to prevent mold.
You can always seal the edges of the coverslip.
Barry



On Tue, Oct 29, 2013 at 1:47 PM, Keyser Gerald T gkey...@uwhealth.orgwrote:

 I'm toying with making my own aqueous mounting media. Although I will not
 likely use this in actual cases, it would still be nice. Aqueous mounting
 media is kind of expensive. But, there are some questions that need to be
 answered.

 The Slides need to be kept for 10 years.

 - Will the adhesive yellow with time?

 - Will the adhesive continue to have complete coverage over time. Over
 drying? Cracking?

 - Since the mounting media is made from glycerin, gelatin, and water, will
 microbial invasion of the slides be a problem? I can add a anti-microbial
 agent to the recipe.

 Anyone have long term experience with this recipe?

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Re: [Histonet] Re: India Ink for marking surgical margin borders

[Barry Rittman]

Bob
Good point - it is true that they are not in particles when used as a
solution - but the colors can also be seen by visible light as well as by
fluorescence.
When I used to mark  biopsy specimens it was mainly to allow correct
orientation of the block in the wax and to allow the  marked area to be
seen when orienting and cutting the block rather than visualizing in a
section.
Barry



On Sat, Oct 12, 2013 at 12:15 PM, Bob Richmond rsrichm...@gmail.com wrote:

 Barry Rittman observes: One group of dyes that I do not believe has ben
 mentioned is the Procion group of dyes. These are chloro-s-triazone dyes
 that are used extensively in textile dyeing and are resistant to all the
 solvents of which I am aware and will survive processing through paraffin
 wax.  Readily available at many art stores. They come in a wide variety of
 colors. They interact with proteins. Their drawback is that many are
 fluorescent (and were originally intended as markers for hard tissue growth
 due to their solvent resistance).

 But if they aren't in insoluble particles, then I won't be able to see them
 under the microscope!

 Bob Richmond
 Samurai Pathologist
 Maryville TN
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Re: [Histonet] India Ink for inking surgical margin borders

[Barry Rittman]

One group of dyes that I do not believe has ben mentioned is the Procion
group of dyes. These are chloro-s-triazone dyes that are used extensively
in textile dyeing and are resistant to all the solvents of which I am aware
and will survive processing through paraffin wax.  Readily available at
many art stores.
They come in a wide variety of colors .   They interact with proteins.
Their drawback is that many are fluorescent (and were originally intended
as markers for hard tissue growth due to their solvent resistance).
Barry


On Fri, Oct 11, 2013 at 10:44 AM, Blazek, Linda 
lbla...@digestivespecialists.com wrote:

 LOL  It's Friday...  I can't resist!  You SUED India Ink?

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
 wanda.sm...@hcahealthcare.com
 Sent: Friday, October 11, 2013 11:38 AM
 To: ecre...@cmblabs.com; histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] India Ink for inking surgical margin borders

 We sued India Ink from an art supply store for years.

 WANDA G. SMITH, HTL(ASCP)HT
 Pathology Supervisor
 TRIDENT MEDICAL CENTER
 9330 Medical Plaza Drive
 Charleston, SC  29406
 843-847-4586
 843-847-4296 fax

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 the number listed.

 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu [mailto:
 histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ed Crespo
 Sent: Thursday, October 10, 2013 12:53 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] India Ink for inking surgical margin borders

 I normally purchase India Ink from one of our vendors, but know it's also
 sold and used at artist supply shops.  Does anyone know if I can used the
 artist india ink for Pathology use?  Really, the only issue would be if the
 ink stays on the tissue during processing right? Please advise.

 Ed Crespo, CT(ASCP)
 Anatomic Pathology Manager
 Safety Officer


 10700 Walker Street
 Cypress, CA 90630
 phone: 714 880.3330
 fax: 714 816.1511
 email: ecre...@cmblabs.com
 cmblabs.com


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