RE: [Histonet] IHC staining weaker in...

From:"Garth Jerome"



Hello

It sounds to me like inadequate fixation of the tissue. The fact that
peripheral staining is good and gets worse towards the middle could indicate
poor penetration of fixative. What is your tissue processing programme?
What fixative are you using?

Regards

Garth Jerome
Histologist
Apollo Scientific cc
Telephone : 27-11-466 7666
Facsimile : 27-11-466 7672
Cell phone : 084 504 1101
Email : garth@apollosci.co.za
-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Department
of Pathology
Sent: 06 September 2007 05:48 PM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] IHC staining weaker in...

Dear Participants, our diagnostic pathology lab has a problem. The paraffin
sections 
are stained weakly at the center but better at the periphery of the
sections. What 
could be the reason.??... thanks a lot.. Dr Nasuhi Engin Aydin, malatya
Inonu 
University Hospital, Turkey 44365.


---------- Original Message -----------
From: histonet-request@lists.utsouthwestern.edu
To: histonet@lists.utsouthwestern.edu
Sent: Thu,  6 Sep 2007 14:51:24 +0300 (EEST)
Subject: Histonet Digest, Vol 46, Issue 6

> Send Histonet mailing list submissions to
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> When replying, please edit your Subject line so it is more specific
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> 
> Today's Topics:
> 
>    1. Xylene (themagoos)
>    2. RE:The great apoptosis stains debate (TUNEL, Caspases,	etc)
>       (Melissa Gonzalez)
>    3. Re: RDO decalcifier (Robert Richmond)
>    4. Re: IHC anti-cleaved caspase 3 (Carl Hobbs)
>    5. avian sperm cell topography (melissah rowe)
>    6. Re: IHC anti-cleaved caspase 3 (Thomas Pier)
>    7. Re: IHC anti-cleaved caspase 3 (Cheryl Cross)
>    8. Fwd: Users of Lab Vision autostainer (Victoria Baker)
>    9. Re: Microwaves VENDOR RESPONSE (Phil McArdle)
>   10. Re: Xylene (Rene J Buesa)
>   11. RE: IHC anti-cleaved caspase 3 (Liz Chlipala)
>   12. Freezing mouse testes for frozen sections
>       (Sarah Clatterbuck Soper)
>   13. Olympus BX40 (Mike Pence)
>   14. Look for a used stereoscope (Yu, Jian)
>   15. RE: RDO decal (Tony Henwood)
>   16. PTH antibody reacted with dog tissue... (ChoiUl Soo)
>   17. Floater sources (Michelle McCoy)
>   18. RE: Xylene (Kemlo Rogerson)
>   19. RE: Floater sources (Kemlo Rogerson)
>   20. Fwd: Users of Lab Vision autostainer (Victoria Baker)
>   21. RE: Xylene (Joseph Kapler)
>   22. Unsubscribe (Malam Jacqueline)
>   23. Re: Floater sources (Joe Nocito)
> 
> ----------------------------------------------------------------------
> 
> Message: 1
> Date: Wed, 05 Sep 2007 10:21:51 -0700
> From: "themagoos" 
> Subject: [Histonet] Xylene
> To: histonet@lists.utsouthwestern.edu
> Message-ID: <46dee5af.14b.450a.1030817715@rushmore.com>
> Content-Type: text/plain; charset="iso-8859-1"
> 
> Can somebody tell me if there are any effects on tissue if
> there is prolonged processing time in xylene?
> 
> Jason McGough HT(ASCP)
> Clinical Laboratory of the Black Hills
> Account Representative - Anatomic Pathology 
> 2805 5th Street
> Rapid City, SD 57701
> 605-343-2267
> jmcgough@clinlab.com
> 
> ------------------------------
> 
> Message: 2
> Date: Wed, 5 Sep 2007 11:30:38 -0700
> From: "Melissa Gonzalez" 
> Subject: [Histonet] RE:The great apoptosis stains debate (TUNEL,
> 	Caspases,	etc)
> To: 
> Message-ID:
> 	<2884B897182A1D438C7BA24B9A8F94A20E701D@hqsvr01mail.cgi.com>
> Content-Type: text/plain;	charset="iso-8859-1"
> 
> Hi all, 
> I have been around this mess a few years back, I gave up on TUNEL long ago
for 
> the various reasons mentioned on the list recently.
> 
> Per investigators requests, I have tried several ways to demonstrate cell=20
> death, however methods such as PI or Annexin staining are more suitable
for 
> whole cells, not tissue sections. I have also not had any luck staining
for 
> Cytochrome C.
> 
> We routinely use R&D Systems rabbit anti human/mouse active Caspase3 using
a 
> high pH (EDTA) based Ag retrieval in a steamer (enzymes do not work).
Works 
> like a charm.
> 
> Any other suggestions? I know this topic has been thrown around a lot, but

> these discussions after all, help us get our jobs done more proficiently.
> 
> Melissa
> 
> Melissa A. Gonzlez Edick
> R&D, Cell Genesys Inc.
> 500 Forbes Blvd
> South San Francisco, CA 94080
> p(650) 266-3168
> f (650) 266-3080
> 
> "It's not enough to believe what you see, you must also understand what
you 
> see." -Leonardo Da Vinci
> ------------------------------
> 
> Message: 6
> Date: Tue, 4 Sep 2007 15:18:39 -0400
> From: Cheryl Cross 
> Subject: Re: [Histonet] TUNEL, Formalin, DNA strand breaks
> To: JR R 
> Cc: histonet@lists.utsouthwestern.edu
> Message-ID: <4C1E074D-064A-45D1-BD32-13D7E2222B26@aol.com>
> Content-Type: text/plain;	charset=US-ASCII;	delsp=yes;
format=flowed
> 
> Hi all -
> 
> I have been round and round with this issue myself; I was basically  
> told by people who've attempted it that TUNEL on FFPE sections is not  
> specific enough due to the formalin issue.
> 
> If you are trying to nail apoptosis, what about anti-active  
> caspase-3? mind you, that staining can be a bit of a booger too, but  
> it should be more sensitive and specific for apoptosis (no references  
> to offer, i'm just repeating what i have been told).
> 
> Cheryl Cross, DVM, Dipl. ACVP
> Researcher
> University Corporation for Atmospheric Research
> College of Veterinary Medicine
> University of Tennessee Department of Pathology
> 2407 River Drive, Room A201
> Knoxville, TN 37996-4542
> (423) 967-2724
> fax: 865-974-5616
> ccross@ucar.edu
> ------------------------------
> 
> Message: 5
> Hi Ray,
> 
> Sheesh-- I read (and then lost) the paper maybe 5-7 years ago, back when I
was 
> tearing my hair out over high background, or false positives for TUNEL
stain 
> in formalin fixed, paraffin embedded arterial sections.  I can
troubleshoot 
> any immunostain, but the TUNEL and ISEL assays bedeviled me.
> 
> The paper was titled something to the effect of "formaldehyde causes
single 
> and double stranded DNA breaks," and was pretty emphatic.  I'll look for
the article.
> 
> I think the assay could work in whole cells for flow, or maybe in frozen=20
> tissue.  My hunch at this point is that this is one of those rare cases
where 
> lots and lots of peer reviewed articles are just plain wrong.
> 
> Hey, if someone has a good protocol, I'd be willing to try it out, and I
would 
> be delighted if I turn out to be mistaken.  For now, I think TUNEL is the=20
> assay of the beast.
> 
> Oh, here is a thought--say you are looking at a 5 micron section through a

> nucleus.  The microtome blade pretty much had to create a lot of double=20
> stranded DNA breaks, no?
> 
> Jerry L. Ricks
> Research Scientist
> U.W. Medicine at South Lake Union
> 815 Mercer Street
> Seattle, WA 98109
> (206)-685-7190
> 
> From: koellingr@comcast.netTo: rosenfeldtek@hotmail.com; 
> histonet@lists.utsouthwestern.eduSubject: RE: [Histonet] TUNELDate: Fri,
31 
> Aug 2007 23:17:03 +0000
> 
> Jerry,
> Could you expand on or give references to formalin causing DNA strand
breaks?  
> Double strand breaks, single strand, blunt end, overhanging?  My pile of=20
> papers and having done TUNEL for years says that formalin fixation is a
very 
> good technique for TUNEL and many peer-reviewed articles in which TUNEL is

> used as a technique, use formalin fixation and how can that be if formalin
is 
> causing strand breaks?  In fact one paper I'm looking at says that
extended (5-
> 7 weeks in formalin) fixation causes loss of TUNEL signal.  If formalin is

> causing breaks, you would assume that TUNEL pos signals would increase
with 
> extended formalin fixation.  Even the use of the monoclonal antibody
F7-26,
>  for single stranded DNA, touts formalin fixation for their claims of 
> discriminating apoptosis from necrosis.
> 
> The question asks about extended alcohol fixation but your answer is
possibly 
> a lot of false positives because formalin causes DNA breaks.  Does this
imply 
> that alcohol won't?  Have done a lot of TUNEL on alcohol fixed samples.
True 
> I couldn't pretreat them and handle them they way I would handle FFPE
tissue.  
> Also true that we could argue specificity and ability or not to
discriminate 
> apoptosis from necrosis for quite a while.  But if formalin itself is
causing 
> the breaks in DNA, I and a lot of people are in big trouble with our
science 
> projects and experiments.  Also I can't envision why 2 cells are showing
TUNEL 
> positivity while 2 of the same type of cells right next to them (and
getting 
> the same formalin fix), are absolutely clean and there is no background?
> 
> Thanks for any information you can provide.
> 
> Ray Koelling
> PhenoPath Laboratories
> Seattle, WA
> 
> ------------------------------
> 
> Message: 3
> Date: Wed, 5 Sep 2007 14:56:14 -0400
> From: "Robert Richmond" 
> Subject: [Histonet] Re: RDO decalcifier
> To: histonet@lists.utsouthwestern.edu
> Message-ID:
> 	
> Content-Type: text/plain; charset=ISO-8859-1
> 
> I don't see how your hazmats people can tell you what to do to dispose
> of RDO, since nobody has any idea what's in it. It's a murky yellow
> liquid that continuously throws a black sediment that has to be
> filtered out if you're to see your specimen in it. It's sort of the
> ultimate secret formula, and I've always wondered why it's so popular
> - I've had to use it many different places in my travels.
> 
> I'd suggest either an ordinary proprietary decalcifier that's a clear
> liquid, probably hydrochloric acid - or else save money and dilute
> your own hydrochloric acid. You can re-use it for a while. When the
> time comes to dispose of it, it can be safely diluted with a lot of
> water and put down the drain.
> 
> Remember - it's been stressed on this list many times - that "Decal"
> is a brand name still in use, and that other brands of decalcifying
> solution should not be called Decal.
> 
> Bob Richmond
> Samurai Pathologist (never a decalcified Carmelite)
> Knoxville TN
> 
> ------------------------------
> 
> Message: 4
> Date: Wed, 5 Sep 2007 20:29:36 +0100
> From: "Carl Hobbs" 
> Subject: Re: [Histonet] IHC anti-cleaved caspase 3
> To: "Histonet" 
> Message-ID: <001801c7eff3$18894850$4101a8c0@carlba65530bda>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> 	reply-type=original
> 
> Hi.
> 
> Recent  Cleaved caspase 3 posts have not mentioned the antibody details: I

> would be grateful for the source of these Abs.
> Carl
> 
> ------------------------------
> 
> Message: 5
> Date: Wed, 5 Sep 2007 14:36:50 -0500
> From: melissah rowe 
> Subject: [Histonet] avian sperm cell topography
> To: histonet@lists.utsouthwestern.edu
> Message-ID: 
> Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed
> 
> Hi,
> 
> I am a PhD student working on a behavioral ecology project on  
> Australian fairy-wrens. I am currently trying to find out if I can  
> determine the dimensions of discrete sections of individual sperm  
> cells, i.e. acrosome length, nucleus length, midpiece length and tail  
> length using the material I currently have available. I have prepared  
> slide smears of sperm that has been previously stained with an eosin- 
> nigrosin stain (to determine cell viability) and would like to be  
> able to somehow stain different segments of the sperm so that I can  
> identify and measure them using an ocular micrometer on a bright  
> field or phase contrast microscope set up. It is my understanding  
> that I cannot use fluorescent stains given that I have already used a  
> colormetric stain (the eosin-nigrosin). If anyone has suggestions for  
> ways to identify discrete regions of avian (passerine) sperm cells I  
> would be very grateful to hear from you.
> 
> Many thanks in advance,
> melissah
> 
> -----
> melissah rowe
> 
> PhD candidate
> Department of Ecology & Evolution
> University of Chicago
> E. 57th Street, Chicago, IL, 60637
> ph: +1 773-702-3070
> fax: + 1 773-702-9740
> 
> email: melissah@uchicago.edu
> 
> ------------------------------
> 
> Message: 6
> Date: Wed, 05 Sep 2007 14:39:04 -0500
> From: "Thomas Pier" 
> Subject: Re: [Histonet] IHC anti-cleaved caspase 3
> To: , 
> Message-ID: <46DEBF88020000DF0000A736@gwmail.medicine.wisc.edu>
> Content-Type: text/plain; charset=US-ASCII
> 
> Cell Signalling Technology has a good rabbit monoclonal for Cleaved
Caspase-3.
> 
> Tom Pier
> 
> >>> "Carl Hobbs"  09/05/07 2:29 PM >>>
> Hi.
> 
> Recent  Cleaved caspase 3 posts have not mentioned the antibody details: I

> would be grateful for the source of these Abs.
> Carl
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> ------------------------------
> 
> Message: 7
> Date: Wed, 5 Sep 2007 15:54:35 -0400
> From: Cheryl Cross 
> Subject: Re: [Histonet] IHC anti-cleaved caspase 3
> To: "Thomas Pier" 
> Cc: carl.hobbs@kcl.ac.uk, histonet@lists.utsouthwestern.edu
> Message-ID: 
> Content-Type: text/plain;	charset=US-ASCII;	delsp=yes;
format=flowed
> 
> I will second the Cell Signaling antibody - we have tried antibodies  
> from Promega (which gave tons of background staining)...then via this  
> site i was pointed to Cell Signaling's monoclonal which we tried; the  
> polyclonal actually has been working very well with minimal  
> background (we were getting lots of respiratory epithelium and  
> endothelium lighting up in control animals). I have been using it in  
> mice with this protocol:
> 
> EDTA with PASCAL
> primary antibody 60 minutes, dilution 1:125
> Rabbit envision
> DAB plus
> 
> Hope this helps!
> 
> Cheryl Cross, DVM, Dipl. ACVP
> Researcher
> University Corporation for Atmospheric Research
> College of Veterinary Medicine
> University of Tennessee Department of Pathology
> 2407 River Drive, Room A201
> Knoxville, TN 37996-4542
> (423) 967-2724
> fax: 865-974-5616
> ccross@ucar.edu
> 
> >
> 
> ------------------------------
> 
> Message: 8
> Date: Wed, 5 Sep 2007 15:59:45 -0400
> From: "Victoria Baker" 
> Subject: [Histonet] Fwd: Users of Lab Vision autostainer
> To: "Histo Net list server" 
> Message-ID:
> 	<4f016b690709051259rcdffccet70d7f97ae0457707@mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
> 
> One more try, first one didn't go through it seems!  Thanks
> 
> ---------- Forwarded message ----------
> From: Victoria Baker 
> Date: Sep 5, 2007 9:02 AM
> Subject: Users of Lab Vision autostainer
> To: Histo Net list server 
> 
> Hi
> 
> I'm a new user of the Lab Vision autostainer and I'm looking to see if
> I can find  users in Histo-land that have experience with it.  The
> facility only works with human tissue and all of the antibodies are
> for dx purposes.
> 
> My key questions are as follows:
> How many antibodies is your lab running?
> How many users do you allow?
> How many people do you allow programming rights and at what level of
> supervision are they?
> 
> How many of these antibodies are from Lab Vision?
>         a)  are they concentrates or pre-dilutes?
>         b)  for HIER are you using their PT modules/procedures or
> your own equipment (microwave, steamer, pressure cooker etc) and in
> house designed protocols for retrieval?
>        c) for digestion do you only use their Pro-K or have you
> designed your own in-house methods using other reagents for digestion?
>        d)  do you put your controls on the same slide as the patient?
>        e)  are your controls in-house or commercial?
>        f)  do you have more than one stainer hooked up to one computer
system?
> 
> What Version of software do you currently have on your system?
> 
> Any feed back would be very helpful.
> 
> Thanks in advance.
> 
> Vikki Baker
> Interim Histology Manager
> Mission Hospital System
> Asheville, NC
> 
> ------------------------------
> 
> Message: 9
> Date: Wed, 05 Sep 2007 16:00:47 -0400
> From: Phil McArdle 
> Subject: Re: [Histonet] Microwaves VENDOR RESPONSE
> To: Joe Nocito 
> Cc: histonet@lists.utsouthwestern.edu
> Message-ID: <46DF0AEF.7030203@ebsciences.com>
> Content-Type: text/plain; charset=UTF-8; format=flowed
> 
> Hi Joe:
> 
> Obviously, a microwave vendor hates to hear microwave horror stories, so=20
> again, no argument - even though I'm not privy to details of the fried 
> biopsies in question or what type/vintage of microwave, anyone who's 
> experienced a malfunction involving patient samples doesn't want a 
> repeat performance. And pathology is, must be, risk averse.
> 
> That said, again, any mechanical or electronic equipment can fail, or 
> user error can contribute; just look at the "hang-up" problems with 
> older tissue processors that are now ancient history. It's up to 
> manufacturers to minimize the possibilities of failure, since patient 
> care is at stake. Improvement is therefore a continual, ongoing process.=20
> For example, while for years EBS microwave processors incorporated 
> safety shutdown modes in the event of vent failure, probe failure (open=20
> and closed) and many other component-related issues, about two years ago=20
> we determined that the microwave did not have a comprehensive set of 
> safeguards to deal with user errors, for example, temperature overshoots=20
> caused by too small a container for a given power setting, or failure to=20
> place the temperature probe in solution. So we developed multiple safety=20
> mechanisms to head off user errors of this sort.
> 
> PMM
> -- 
> Phil McArdle
> Microwave Product Manager
> 
> Energy Beam Sciences, Inc.
> 29-B Kripes Rd.
> East Granby, CT 06026
> 
> Tel:  800.992.9037 x 341
> Mobile: 860.597.6796
> Fax: 860.653.0422
> 
> pmcardle@ebsciences.com
> www.ebsciences.com
> 
> Joe Nocito wrote:
> > are you sure it's the willies and not the johnnies?
> > Since the magnetron or whatever it was that fried my tissue, I'd wait=20
> > for the traditional processing. Call me a dinosaur, but I really don't=20
> > like doing special stains in the microwave. The only thing I use a 
> > microwave for at my house is to defrost and reheat stuff (technical 
> > term). I'm sure there are people out there who can cook a 6 course 
> > gourmet meal. My best friend can process all types of tissue from 
> > biopsies to uterus.
> >    As a matter of fact, he was there grossing when something went wrong=20
> > and told me that he's never seen tissue like that before.
> > 
> > JTT
> > ----- Original Message ----- From: "Phil McArdle"

> > To: "Joe Nocito" 
> > Cc: 
> > Sent: Wednesday, September 05, 2007 11:16 AM
> > Subject: Re: [Histonet] Microwaves VENDOR RESPONSE
> > 
> > 
> >> Hi Joe:
> >>
> >> No argument there. I'm painfully aware of both a mindset of "a 
> >> microwave 'should' cost less than $100," and of a dearth of funding 
> >> for pathology in general (popular shows like CSI to the contrary). :-)=20
> >> I'd still suggest that $1749 is a heck of a lot better (and a lot less=20
> >> laughable) than the $18,000 or $30,000 that's widely quoted and 
> >> posted, and it's the exact reason we brought an under-$2000 lab 
> >> microwave to market in the first place.
> >>
> >> One could argue just as convincingly against all kinds of specialized=20
> >> equipment or reagents on the basis of cost, not just microwaves. We 
> >> all know of everything from saliva to cheap rice steamers being used=20
> >> in histo labs, and while they may actually be perfectly serviceable,=20
> >> from the standpoint of repeatability or liability, this kind of thing=20
> >> gives me the willies (and that's a technical term). My yardstick is 
> >> always "what would I be comfortable with if my kid's diagnosis hung in=20
> >> the balance?"
> >>
> >> Healthy debate is good!
> >>
> >> Phil
> >> -- 
> >> Phil McArdle
> >> Microwave Product Manager
> >>
> >> Energy Beam Sciences, Inc.
> >> 29-B Kripes Rd.
> >> East Granby, CT 06026
> >>
> >> Tel:  800.992.9037 x 341
> >> Mobile: 860.597.6796
> >> Fax: 860.653.0422
> >>
> >> pmcardle@ebsciences.com
> >> www.ebsciences.com
> >>
> >>
> >>
> >>
> >>
> >>
> >> Joe Nocito wrote:
> >>> ok, but with the budgets today, many people can't afford a $1749 
> >>> microwave when they can buy one at Walmart, K-Mart, or somewhere else=20
> >>> for $79.
> >>>    Not to make you angry or anything, but I'm wondering how long has=20
> >>> it been since you worked in a lab? Histo's budget is the first one 
> >>> cut in the lab because we are not essential.
> >>> I can't count how many times I fought and fought for my budgets.
> >>>    If I tried to justify a $1749 microwave for special stains, HIER=20
> >>> or whatever, I would have been laughed out the manager's office.
> >>>    Just my 4 cents.
> >>>
> >>> JTT
> >>> ----- Original Message ----- From: "Phil McArdle" 
> >>> 
> >>> To: "Kathleen Boozer" 
> >>> Cc: 
> >>> Sent: Wednesday, September 05, 2007 9:07 AM
> >>> Subject: Re: [Histonet] Microwaves VENDOR RESPONSE
> >>>
> >>>
> >>>> Again, a microwave vendor weighs in (so far I haven't received any=20
> >>>> flames), so read at your own risk. :-)
> >>>>
> >>>> At the risk of sounding overly and overtly commercial, after reading=20
> >>>> post after post of $30,000+ and $18,000 and similarly high figures=20
> >>>> for lab microwaves, I really feel the need to set the record 
> >>>> straight. Depending on the usage requirements, we have laboratory 
> >>>> microwaves as low as $1749 for a "bare bones" model for simple 
> >>>> operations, to mid-priced units, to under $11,000 for a vacuum 
> >>>> equipped microwave processor capable of the +/- 0.5 degree C 
> >>>> temperature control necessary for tissue processing.
> >>>>
> >>>> (I can feel the heat already!)
> >>>>
> >>>> There are many compelling reasons to replace a kitchen microwave 
> >>>> with a lab model; feel free to download, read, and even share with=20
> >>>> colleagues our Microwave Companion at
> >>>>
> >>>> http://www.ebsciences.com/pdf/EBS_MW_COMPANION.pdf
> >>>>
> >>>> Best regards, and see you at NSH,
> >>>>
> >>>> Phil McArdle
> >>>>
> >>>> -- 
> >>>> Phil McArdle
> >>>> Microwave Product Manager
> >>>>
> >>>> Energy Beam Sciences, Inc.
> >>>> 29-B Kripes Rd.
> >>>> East Granby, CT 06026
> >>>>
> >>>> Tel:  800.992.9037 x 341
> >>>> Mobile: 860.597.6796
> >>>> Fax: 860.653.0422
> >>>>
> >>>> pmcardle@ebsciences.com
> >>>> www.ebsciences.com
> >>>>
> >>>> Kathleen Boozer wrote:
> >>>>> What is the best microwave for a small lab using it only for 
> >>>>> heating Bouin's and Silver Nitrate for special stains?  I just 
> >>>>> can't believe I would have to spend $30,000+ or slow down and use a=20
> >>>>> waterbath.
> >>>>>
> >>>>>
> >>>>> _______________________________________________
> >>>>> Histonet mailing list
> >>>>> Histonet@lists.utsouthwestern.edu
> >>>>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>>>
> >>>>
> >>>>
> >>>>
> >>>> I skate to where the puck is going to be, not to where it's been.
> >>>> - Wayne Gretsky
> >>>>
> >>>> You must be the change you want to see in the world.
> >>>> - Mahatma Gandhi
> >>>>
> >>>> NOTE: This message, together with any attachments, is intended only=20
> >>>> for the use of the individual or entity to which it is addressed and=20
> >>>> may contain information that is legally privileged, confidential and=20
> >>>> exempt from disclosure. If you are not the intended recipient, 
> >>>> however, there's not a lot I can do about it, and it was probably my=20
> >>>> mistake anyway. So please do the right thing and make this e-mail go=20
> >>>> away. Thank you.
> >>>>
> >>>> _______________________________________________
> >>>> Histonet mailing list
> >>>> Histonet@lists.utsouthwestern.edu
> >>>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>>
> >>>
> >>> _______________________________________________
> >>> Histonet mailing list
> >>> Histonet@lists.utsouthwestern.edu
> >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> >>
> >>
> >> -- 
> >> Phil McArdle
> >> Microwave Product Manager
> >>
> >> Energy Beam Sciences, Inc.
> >> 29-B Kripes Rd.
> >> East Granby, CT 06026
> >>
> >> Tel:  800.992.9037 x 341
> >> Mobile: 860.597.6796
> >> Fax: 860.653.0422
> >>
> >> pmcardle@ebsciences.com
> >> www.ebsciences.com
> >>
> >> I skate to where the puck is going to be, not to where it's been.
> >> - Wayne Gretsky
> >>
> >> You must be the change you want to see in the world.
> >> - Mahatma Gandhi
> >>
> >> NOTE: This message, together with any attachments, is intended only 
> >> for the use of the individual or entity to which it is addressed and=20
> >> may contain information that is legally privileged, confidential and=20
> >> exempt from disclosure. If you are not the intended recipient, 
> >> however, there's not a lot I can do about it, and it was probably my=20
> >> mistake anyway. So please do the right thing and make this e-mail go=20
> >> away. Thank you. 
> >
> 
> I skate to where the puck is going to be, not to where it's been.
> - Wayne Gretsky
> 
> You must be the change you want to see in the world.
> - Mahatma Gandhi
> 
> NOTE: This message, together with any attachments, is intended only for=20
> the use of the individual or entity to which it is addressed and may 
> contain information that is legally privileged, confidential and exempt=20
> from disclosure. If you are not the intended recipient, however, there's=20
> not a lot I can do about it, and it was probably my mistake anyway. So 
> please do the right thing and make this e-mail go away. Thank you.
> 
> ------------------------------
> 
> Message: 10
> Date: Wed, 5 Sep 2007 13:01:21 -0700 (PDT)
> From: Rene J Buesa 
> Subject: Re: [Histonet] Xylene
> To: themagoos@rushmore.com, histonet@lists.utsouthwestern.edu
> Message-ID: <329330.36844.qm@web61219.mail.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
> 
> Tissues usually become brittle, which difficulties sectioning.
>   Ren J.
> 
> themagoos  wrote:
>   Can somebody tell me if there are any effects on tissue if
> there is prolonged processing time in xylene?
> 
> Jason McGough HT(ASCP)
> Clinical Laboratory of the Black Hills
> Account Representative - Anatomic Pathology 
> 2805 5th Street
> Rapid City, SD 57701
> 605-343-2267
> jmcgough@clinlab.com
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> ---------------------------------
> Boardwalk for $500? In 2007? Ha! 
> Play Monopoly Here and Now (it's updated for today's economy) at Yahoo!
Games.
> 
> ------------------------------
> 
> Message: 11
> Date: Wed, 5 Sep 2007 14:09:02 -0600
> From: "Liz Chlipala" 
> Subject: RE: [Histonet] IHC anti-cleaved caspase 3
> To: "Cheryl Cross" , "Thomas Pier"
> 	
> Cc: carl.hobbs@kcl.ac.uk, histonet@lists.utsouthwestern.edu
> Message-ID:
> 	
> Content-Type: text/plain;	charset="windows-1250"
> 
> I like the cell signaling antibody also, I tried biocare's but did not
have 
> much success with it.  The cell signaling antibody also works with pronase

> digestion, as well as the EDTA pH9 HIER.  We have even used it on bone 
> sections with the pronase digestion. Works in multiple species, I have
used it 
> on human, rat, mouse, guinea pig, porcine and canine.  Our protocol is
similar 
> to Cheryl's.  It’s a bit pricy as antibodies go, but I feel its
worth it.  
> Good lot to lot consistency.  We have probably gone through about 5
different 
> lots, with not changing the protocol at all.  I have a written protocol if

> anyone is interested.
> 
> Liz
> 
> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
> Manager
> Premier Laboratory, LLC
> P.O. Box 18592
> Boulder, CO 80308
> phone (303) 735-5001
> fax (303) 735-3540
> liz@premierlab.com
> www.premierlab.com
> 
> Ship to Address:
> 
> Premier Laboratory, LLC
> University of Colorado at Boulder
> MCDB, Room A3B40
> Boulder, CO 80309
> 
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-
> bounces@lists.utsouthwestern.edu] On Behalf Of Cheryl Cross Sent:
Wednesday, 
> September 05, 2007 2:01 PM To: Thomas Pier Cc: carl.hobbs@kcl.ac.uk; 
histonet@lists.utsouthwestern.edu
> Subject: Re: [Histonet] IHC anti-cleaved caspase 3
> 
> I will second the Cell Signaling antibody - we have tried antibodies from=20
> Promega (which gave tons of background staining)...then via this site i
was 
> pointed to Cell Signaling's monoclonal which we tried; the polyclonal
actually 
> has been working very well with minimal background (we were getting lots
of 
> respiratory epithelium and endothelium lighting up in control animals). I
have 
> been using it in mice with this protocol:
> 
> EDTA with PASCAL
> primary antibody 60 minutes, dilution 1:125 Rabbit envision DAB plus
> 
> Hope this helps!
> 
> Cheryl Cross, DVM, Dipl. ACVP
> Researcher
> University Corporation for Atmospheric Research College of Veterinary
Medicine 
> University of Tennessee Department of Pathology 2407 River Drive, Room
A201 
> Knoxville, TN 37996-4542
> (423) 967-2724 fax: 865-974-5616 ccross@ucar.edu
> 
> >
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> No virus found in this incoming message.
> Checked by AVG Free Edition. 
> Version: 7.5.485 / Virus Database: 269.13.5/990 - Release Date: 9/4/2007
10:36 
> PM
> 
> No virus found in this outgoing message.
> Checked by AVG Free Edition. 
> Version: 7.5.485 / Virus Database: 269.13.5/990 - Release Date: 9/4/2007
10:36 
> PM
> 
> ------------------------------
> 
> Message: 12
> Date: Wed, 05 Sep 2007 16:13:36 -0400
> From: Sarah Clatterbuck Soper 
> Subject: [Histonet] Freezing mouse testes for frozen sections
> To: histonet@lists.utsouthwestern.edu
> Message-ID: <46DF0DF0.2030306@ciwemb.edu>
> Content-Type: text/plain; charset=ISO-8859-1; format=flowed
> 
> Hi all,
> 
> I've started attempting to section unfixed frozen mouse testes in order=20
> to placate a specific antibody we have to use.  I am new to frozen 
> sections and I'm having trouble with the testes cracking when I freeze 
> them.  I've tried both freezing in isopentane cooled on liquid nitrogen=20
> and an acetone/dry ice slurry.  Either way the testes crack, usually one=20
> big crack end to end.  Doesn't seem to be as much of a problem with our=20
> mutant testes, which are about 1/3 the size of wild-type.  I wish I 
> could just trim the wild-type down to a smaller size, but obviously 
> that's not an option!
> 
> Any recommendations?
> 
> Thanks so much!
> 
> Sarah
> 
> ------------------------------
> 
> Message: 13
> Date: Wed, 5 Sep 2007 15:49:48 -0500
> From: "Mike Pence" 
> Subject: [Histonet] Olympus BX40
> To: 
> Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C701@IS-E2K3.grhs.net>
> Content-Type: text/plain;	charset="us-ascii"
> 
> Need some help,
> 
> I am looking for a 60x objective for an Olympus microscope BX40.
> Would anyone know where I might get a used one or if they even make one
> this size for this scope?
> 
> Thanks,
> Mike
> 
> ------------------------------
> 
> Message: 14
> Date: Wed, 5 Sep 2007 16:58:12 -0400
> From: "Yu, Jian" 
> Subject: [Histonet] Look for a used stereoscope
> To: 
> Message-ID:
>
<7E0A77BFEB9A1E47A63F978E7116821F07986D6D@1upmc-msx11.acct.upmchs.net>
> Content-Type: text/plain;	charset="us-ascii"
> 
> Does anyone know a good place to get a used stereoscope?  I plan to use
> it to examine intestinal tumors in mice.
> 
> Thanks a lot for your information.
> 
> *******************************************************************
> Jian Yu, Ph.D.
> University of Pittsburgh Cancer Institute
> Hillman Cancer Center Research Pavilion
> Office Suite 2.26h
> 5117 Centre Avenue, Pittsburgh, PA 15213
> *******************************************************************
> 
> ------------------------------
> 
> Message: 15
> Date: Thu, 6 Sep 2007 09:44:45 +1000
> From: "Tony Henwood" 
> Subject: RE: [Histonet] RDO decal
> To: "Rene J Buesa" , "RENEE FISHER"
> 	,	
> Message-ID: 
> Content-Type: text/plain; charset="iso-8859-1"
> 
> There is also the issue of EDTA, which is common in many RDO formulations.
> Checking one MSDS for EDTA reveals:
> 
> Ecological Information
>   Environmental Fate: 
>   When released into the soil, this material is expected to leach into 
>   groundwater. When released into the soil, this material may biodegrade
to a 
>   moderate extent. When released into the soil, this material is not
expected 
> to   evaporate significantly. When released into water, this material is
not   
> expected to evaporate significantly. This material is not expected to  
>  significantly bioaccumulate. When released into the air, this material is

> expected to be readily degraded by photolysis.   Environmental Toxicity:

> This material is not expected to be toxic to aquatic life. The
LC50/96-hour  
>  values for fish are over 100 mg/l.
> 
> So neutralisation may not be required.
> If anyone has info to the contrary please advise.
> 
> Regards
> 
> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
> Laboratory Manager & Senior Scientist
> The Children's Hospital at Westmead,
> Locked Bag 4001, Westmead, 2145, AUSTRALIA.
> Tel: 612 9845 3306
> Fax: 612 9845 3318
> 
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-
> bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent:
Thursday, 6 
> September 2007 1:07 AM To: RENEE FISHER; histonet@lists.utsouthwestern.edu

> Subject: Re: [Histonet] RDO decal
> 
> Rene:
>   I have not heard of neutralizing RDO, but it would make sense if you
want to 
> be "gentle on your sewer system" BUT prepare to a large emission of carbon

> dioxide when attempting to neutralize it with baking soda.  RDO + baking
soda 
> (or sodium bicarbonate) will produce water, salt with the acid in RDO 
> (probably sodium chloride or common salt), and carbon dioxide in 
> stoichiometrical amounts (1 CO2 per every 1 NaCl).  Therefore that 
> neutralization has to take place in a fumes hood, and you will have to
decide 
> which is worst: delivering acid to the sewer system, or carbon dioxide
(the 
> Greenhouse gas per excellence) to the atmosphere.  It will be "your call".

>  Ren J.
> 
> RENEE FISHER  wrote:
>   Has anyone heard of neutralizing RDO with baking soda to P.H. 7.0. Our
Histo 
> lab had a hazardous waste assessment, and the consultant suggested we not=20
> throw the RDO down the drain but that we either collect it for waste
removal 
> or neutralize it with baking soda to p.h. 7.0. I have not heard of doing
this 
> and do not know of any procedure for it, everyone, anyone's help will be=20
> greatly appreciated.
> 
> Thanks,
> Renee'
> 
> 
____________________________________________________________________________
___________
> 
> This email may contain confidential protected health information and/or=20
> proprietary information belonging to the sender that is legally privileged

> under local, state, or federal law. This information is intended only for
the 
> use of the individual or individuals who have received this. The
authorized 
> recipient of this information is prohibited from disclosing this
information 
> to any other party unless required to do so by law. If you are not the 
> intended recipient, you are hereby notified that any disclosure, copying,
>  distribution, or action taken in reliance on the contents of this email
is 
> strictly prohibited. If you have received this email in error, please
notify 
> the sender immediately to arrange for the disposal of this information.
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> ---------------------------------
> Take the Internet to Go: Yahoo!Go puts the Internet in your pocket: mail,
news,
>  photos & more. _______________________________________________ Histonet=20
> mailing list Histonet@lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> *********************************************************************
> This email and any files transmitted with it are confidential and intended

> solely for the use of the individual or entity to whom they are addressed.
If 
> you are not the intended recipient, please delete it and notify the
sender.
> 
> Views expressed in this message and any attachments are those of the 
> individual sender, and are not necessarily the views of The Children's 
> Hospital at Westmead
> 
> This note also confirms that this email message has been
> virus scanned and although no computer viruses were detected, The
Childrens 
> Hospital at Westmead accepts no liability for any consequential damage 
> resulting from email containing computer viruses. 
**********************************************************************
> 
> ------------------------------
> 
> Message: 16
> Date: Thu, 6 Sep 2007 11:23:00 +0900
> From: ChoiUl Soo 
> Subject: [Histonet] PTH antibody reacted with dog tissue...
> To: "histonet@lists.utsouthwestern.edu"
> 	
> Message-ID: 
> Content-Type: text/plain; charset="ks_c_5601-1987"
> 
> Hi Histonetters,
> 
> I am interested in anti PTH antibody reacted with dog tissue.
> If anyone has successful experience with any PTH antibody with dog tissue,
> please tell me one.
> 
> I have searched through the internet, and results came back with abcam and

> SantaCruz PTH antibodies predicted to react with them. But they are not
sure 
> on it, just predicted on the basis of sequence homology. 
> (abcam say 94% homology with human) I don't have good exprience with Santa

> Cruz, and never used one by Abcam. Should I rely on it, or find another
one?
> 
> Let me hear your experience.
> 
> I would appreciate your advice or comment on this.
> 
> Thank you~.
> 
> Ul Soo Choi, DVM, PhDKRF priority zoonotic disease research institute,
College 
> of Veterinary Medicine, Seoul National University, Shilim9 dong, Gwanakgu,

> Seoul, Korea 151-742Tel. 82-02-880-8688 Mobile. 82-016-9228-8634Fax.
82-02-880-
> 8662
> 
> _________________________________________________________________
>  ۷ι θ, Windows Live Space!  
> http://www.spaces.live.com
> 
> ------------------------------
> 
> Message: 17
> Date: Thu, 6 Sep 2007 01:57:04 -0400
> From: "Michelle McCoy" 
> Subject: [Histonet] Floater sources
> To: "histonet@lists.utsouthwestern.edu"
> 	
> Message-ID:
> 	<25355ef80709052257i32794198gd5a64b94758cc1ea@mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
> 
> I was reading some of the old archived messages on floaters, and being
> somewhat new to the field of histotechnology was curious about how
floaters
> can be attributed to a particular tech who performed work on the block.
For
> example, couldn't other sources of contamination be from the automatic
> stainer, processor carryover (cassette not completely closed/or if closed,
> friable tissue through the slats eg if sponge not used on larger
specimen).
> In some previous labs I've worked in- floaters were quickly attributed to
> the cutter, embedder or grosser and might result in a "write up". If the
> floater is seen in the block how can you differentiate if it came from the
> grosser carryover/embedder carryover/processing carryover/unsigned
> re-embedder/or even possibly even client carry over between patients or
> different specimen types of the same patient.
> And if it is not in the block --differentiating between the
cutter/automatic
> stainer (I've seen specks of tissue debris in automatic stainers/ sections
> falling off the slides and into the reagents etc). Obviously all should be
> done to minimize the factors, but I'm just not clear how a single source
is
> pinpointed and potentially blamed for the event (depending on the lab
> policy). Thanks for any ideas on this.
> 
> ------------------------------
> 
> Message: 18
> Date: Thu, 6 Sep 2007 08:07:38 +0100
> From: "Kemlo Rogerson" 
> Subject: RE: [Histonet] Xylene
> To: , 
> Message-ID:
>
<86ADE4EB583CE64799A9924684A0FBBF0222EC6C@wahtntex2.waht.swest.nhs.uk>
> Content-Type: text/plain;	charset="us-ascii"
> 
> Can somebody tell me if there are any effects on tissue if there is
> prolonged processing time in xylene?
> 
> Jason McGough HT(ASCP)
> Clinical Laboratory of the Black Hills
> Account Representative - Anatomic Pathology 
> 2805 5th Street
> Rapid City, SD 57701
> 605-343-2267
> jmcgough@clinlab.com
> 
> Classically it is said to harden, plus more lipids could be removed.
> Personally I'm equivocal about that thought; if properly fixed prior to
> processing I would have thought the hardening effects of xylene were
> much less than that of the coagulant fixative ethanol. If you do have
> hard tissue then there is a restorative fluid one can use which I think
> has oil of cedarwood in it but I don't know the formula off hand. I know
> it works cos when I was a pup I used it sometimes.
> 
> Kemlo Rogerson
> Pathology Manager
> DD   01934 647057 or extension 3311
> Mob 07749 754194; Pager 07659 597107;
> 
> Sunshine is delicious, rain is refreshing, wind braces us up, snow is
> exhilarating; there is really no such thing as bad weather, only
> different kinds of good weather. --John Ruskin
> 
> This e-mail is confidential and privileged. If you are not the intended
> recipient please accept my apologies; please do not disclose, copy or
> distribute information in this e-mail or take any action in reliance on
> its contents: to do so is strictly prohibited and may be unlawful.
> Please inform me that this message has gone astray before deleting it.
> Thank you for your co-operation
> 
> ------------------------------
> 
> Message: 19
> Date: Thu, 6 Sep 2007 08:17:36 +0100
> From: "Kemlo Rogerson" 
> Subject: RE: [Histonet] Floater sources
> To: "Michelle McCoy" ,
> 	
> Message-ID:
>
<86ADE4EB583CE64799A9924684A0FBBF0222EC6D@wahtntex2.waht.swest.nhs.uk>
> Content-Type: text/plain;	charset="us-ascii"
> 
> I was reading some of the old archived messages on floaters, and being
> somewhat new to the field of histotechnology was curious about how
> floaters can be attributed to a particular tech who performed work on
> the block. For example, couldn't other sources of contamination be from
> the automatic stainer, processor carryover (cassette not completely
> closed/or if closed, friable tissue through the slats eg if sponge not
> used on larger specimen). In some previous labs I've worked in- floaters
> were quickly attributed to the cutter, embedder or grosser and might
> result in a "write up". If the floater is seen in the block how can you
> differentiate if it came from the grosser carryover/embedder
> carryover/processing carryover/unsigned re-embedder/or even possibly
> even client carry over between patients or different specimen types of
> the same patient. And if it is not in the block --differentiating
> between the cutter/automatic stainer (I've seen specks of tissue debris
> in automatic stainers/ sections falling off the slides and into the
> reagents etc). Obviously all should be done to minimize the factors, but
> I'm just not clear how a single source is pinpointed and potentially
> blamed for the event (depending on the lab policy). Thanks for any ideas
> on this.
> 
> You are exactly correct, floaters can be attributed to a variety of
> causes, processing machines that aren't regularly changed, transfer on
> the cutting up forceps, on the waterbath and on the forceps of the
> embedder. If the floater was from a block that was cut, embedded, cut up
> before the section with the floater then the culprit is obvious. If the
> section was cut by someone who didn't cut the block from which the
> floater floated, then the culprit is the embedder, machine,or cutter up.
> Realisticaly I would have thought that most floaters would be from the
> embedder, cutter up, then sectioner as the latter has the greater
> likelihood of seeing the error of his/ her ways. Waxy forceps of messy
> forceps, in my experience are the usual culprit but in some instances,
> necrotic tumours can shed cells in the processor and even the stainer.
> 
> Floaters are usually obvious as they tend to be at a different level
> than the tissue section and I'm afraid they are a fact of life. You can
> reduce the incidence but sadly never eradicate them.
> 
> Kemlo Rogerson
> Pathology Manager
> DD   01934 647057 or extension 3311
> Mob 07749 754194; Pager 07659 597107;
> 
> Sunshine is delicious, rain is refreshing, wind braces us up, snow is
> exhilarating; there is really no such thing as bad weather, only
> different kinds of good weather. --John Ruskin
> 
> This e-mail is confidential and privileged. If you are not the intended
> recipient please accept my apologies; please do not disclose, copy or
> distribute information in this e-mail or take any action in reliance on
> its contents: to do so is strictly prohibited and may be unlawful.
> Please inform me that this message has gone astray before deleting it.
> Thank you for your co-operation
> 
> ------------------------------
> 
> Message: 20
> Date: Thu, 6 Sep 2007 04:18:04 -0400
> From: "Victoria Baker" 
> Subject: [Histonet] Fwd: Users of Lab Vision autostainer
> To: histonet 
> Message-ID:
> 	<4f016b690709060118t532d22c1if250116e0a6c9af1@mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
> 
> ---------- Forwarded message ----------
> From: Victoria Baker 
> Date: Sep 5, 2007 9:02 AM
> Subject: Users of Lab Vision autostainer
> To: Histo Net list server 
> 
> Hi
> 
> I'm a new user of the Lab Vision autostainer and I'm looking to see if
> I can find  users in Histo-land that have experience with it.  The
> facility only works with human tissue and all of the antibodies are
> for dx purposes.
> 
> My key questions are as follows:
> How many antibodies is your lab running?
> How many users do you allow?
> How many people do you allow programming rights and at what level of
> supervision are they?
> 
> How many of these antibodies are from Lab Vision?
>         a)  are they concentrates or pre-dilutes?
>         b)  for HIER are you using their PT modules/procedures or
> your own equipment (microwave, steamer, pressure cooker etc) and in
> house designed protocols for retrieval?
>        c) for digestion do you only use their Pro-K or have you
> designed your own in-house methods using other reagents for digestion?
>        d)  do you put your controls on the same slide as the patient?
>        e)  are your controls in-house or commercial?
>        f)  do you have more than one stainer hooked up to one computer
system?
> 
> What Version of software do you currently have on your system?
> 
> Any feed back would be very helpful.
> 
> Thanks in advance.
> 
> Vikki Baker
> Interim Histology Manager
> Mission Hospital System
> Asheville, NC
> 
> ------------------------------
> 
> Message: 21
> Date: Thu, 6 Sep 2007 02:28:57 -0600
> From: "Joseph Kapler" 
> Subject: RE: [Histonet] Xylene
> To: "Rene J Buesa" , ,
> 	
> Message-ID: 
> Content-Type: text/plain;	charset="iso-8859-1"
> 
> Tissues processed in Xylene (especially over long periods of time) the
> tissue becomes very brittle.
> 
> Hope that is the response you were looking for.
> 
> Joseph "DarkWolfe" Kapler
> I'm an outsider outside of everything
> I'm an outsider outside of everything
> I'm an outsider outside of everything
> Everything you know. Everything you know
> It disturbs me so
> 
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rene J
> Buesa
> Sent: Wednesday, September 05, 2007 14:01
> To: themagoos@rushmore.com; histonet@lists.utsouthwestern.edu
> Subject: Re: [Histonet] Xylene
> 
> Tissues usually become brittle, which difficulties sectioning.
>   Ren J.
> 
> themagoos  wrote:
>   Can somebody tell me if there are any effects on tissue if
> there is prolonged processing time in xylene?
> 
> Jason McGough HT(ASCP)
> Clinical Laboratory of the Black Hills
> Account Representative - Anatomic Pathology
> 2805 5th Street
> Rapid City, SD 57701
> 605-343-2267
> jmcgough@clinlab.com
> 
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> ---------------------------------
> Boardwalk for $500? In 2007? Ha!
> Play Monopoly Here and Now (it's updated for today's economy) at Yahoo!
> Games.
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> No virus found in this incoming message.
> Checked by AVG Free Edition.
> Version: 7.5.485 / Virus Database: 269.13.5/988 - Release Date: 9/4/2007
> 09:14
> 
> No virus found in this outgoing message.
> Checked by AVG Free Edition.
> Version: 7.5.485 / Virus Database: 269.13.5/988 - Release Date: 9/4/2007
> 09:14
> 
> ------------------------------
> 
> Message: 22
> Date: Thu, 6 Sep 2007 09:59:13 +0100 
> From: Malam Jacqueline 
> Subject: [Histonet] Unsubscribe
> To: histonet@lists.utsouthwestern.edu
> Message-ID: 
> Content-Type: text/plain
> 
> Please would you  unsubscribe me as I am retiring tomorrow - thanks
> 
> Jacqui malam
> Lancaster
> uk
> 
> DISCLAIMER: This e-mail is confidential and privileged. If you are not the
> intended recipient please accept our apologies; please do not disclose,
copy
> or distribute information in this e-mail or take any action in reliance on
> its contents: to do so is strictly prohibited and may be unlawful. Please
> inform postmaster@rli.mbht.nhs.uk that this message has gone astray before
> deleting it.  Comments or opinions expressed in this email are those of
> their respective contributors only. The views expressed do not represent
the
> views of the Trust, its management or employees. University Hospitals of
> Morecambe Bay NHS Trust is not responsible and disclaims any and all
> liability for the content of comments written within.Thank you for your
> co-operation.
> 
> ------------------------------
> 
> Message: 23
> Date: Thu, 6 Sep 2007 06:31:58 -0500
> From: "Joe Nocito" 
> Subject: Re: [Histonet] Floater sources
> To: "Michelle McCoy" ,
> 	
> Message-ID: <004201c7f079$8f5aa560$0202a8c0@yourxhtr8hvc4p>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> 	reply-type=original
> 
> Michelle,
> first. welcome to world of histology.
> Let's begin at the grossing table- all grossers are trained to wipe off
the 
> table after each case- but that doesn't mean floaters can't happen, but
the 
> incidence is low
> third- embedders should have been taught to clean the embedding area after

> each case and at the end of the day, but this is a good source for 
> contamination. You can see this is the block.
> fourth- the cutters should wipe off their waterbaths after each block.
This 
> probably is the most likely source of contamination. I've inspected some=20
> labs where one waterbath was just covered with previous ribbons, attached
to 
> the side and floating on the waterbath.
> fifth- in my experience, it is highly unlikely that floaters came from the

> stainer just by the shear movement of the slides and water.
> 
> The bottom line is that everyone needs to be neat and clean and be 
> meticulous. Remember, cleanliness is next to Godliness.
> I had a testicular seminoma where no matter what we did, floaters still
were 
> on cervical bxs and endometrial bxs.  The medical and I decided that we=20
> would handle cases like these separately. They were processed, embedded,
cut 
> and stained separately. Once the case was completed, we changed all the=20
> solutions on the tissue processor and stainer. Who ever cut the blocks had

> to dismantle their knife holder and microtome to clean it thoroughly. A
pain 
> in the butt, but was a necessary evil.
> 
> I hope this helped a little. Good luck.
> 
> Joe The Toe
> ----- Original Message ----- 
> From: "Michelle McCoy" 
> To: 
> Sent: Thursday, September 06, 2007 12:57 AM
> Subject: [Histonet] Floater sources
> 
> >I was reading some of the old archived messages on floaters, and being
> > somewhat new to the field of histotechnology was curious about how 
> > floaters
> > can be attributed to a particular tech who performed work on the block.=20
> > For
> > example, couldn't other sources of contamination be from the automatic
> > stainer, processor carryover (cassette not completely closed/or if
closed,
> > friable tissue through the slats eg if sponge not used on larger 
> > specimen).
> > In some previous labs I've worked in- floaters were quickly attributed
to
> > the cutter, embedder or grosser and might result in a "write up". If the
> > floater is seen in the block how can you differentiate if it came from
the
> > grosser carryover/embedder carryover/processing carryover/unsigned
> > re-embedder/or even possibly even client carry over between patients or
> > different specimen types of the same patient.
> > And if it is not in the block --differentiating between the 
> > cutter/automatic
> > stainer (I've seen specks of tissue debris in automatic stainers/
sections
> > falling off the slides and into the reagents etc). Obviously all should
be
> > done to minimize the factors, but I'm just not clear how a single source

> > is
> > pinpointed and potentially blamed for the event (depending on the lab
> > policy). Thanks for any ideas on this.
> > _______________________________________________
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> > Histonet@lists.utsouthwestern.edu
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> 
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> End of Histonet Digest, Vol 46, Issue 6
> ***************************************
------- End of Original Message -------


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