[Histonet] IHC staining weaker in...

From:"Department of Pathology"



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

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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 
> 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 
> 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 
> 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 
> 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 
> 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 
> 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. 
> For example, while for years EBS microwave processors incorporated 
> safety shutdown modes in the event of vent failure, probe failure (open 
> and closed) and many other component-related issues, about two years ago 
> we determined that the microwave did not have a comprehensive set of 
> safeguards to deal with user errors, for example, temperature overshoots 
> caused by too small a container for a given power setting, or failure to 
> place the temperature probe in solution. So we developed multiple safety 
> 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 
> > for the traditional processing. Call me a dinosaur, but I really don't 
> > 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 
> > 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). :-) 
> >> I'd still suggest that $1749 is a heck of a lot better (and a lot less 
> >> 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 
> >> equipment or reagents on the basis of cost, not just microwaves. We 
> >> all know of everything from saliva to cheap rice steamers being used 
> >> in histo labs, and while they may actually be perfectly serviceable, 
> >> from the standpoint of repeatability or liability, this kind of thing 
> >> 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 
> >> 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 
> >>> for $79.
> >>>    Not to make you angry or anything, but I'm wondering how long has 
> >>> 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 
> >>> 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 
> >>>> flames), so read at your own risk. :-)
> >>>>
> >>>> At the risk of sounding overly and overtly commercial, after reading 
> >>>> post after post of $30,000+ and $18,000 and similarly high figures 
> >>>> 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 
> >>>> 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 
> >>>>> 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 
> >>>> for the use of the individual or entity to which it is addressed and 
> >>>> may contain information that is legally privileged, confidential and 
> >>>> 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 
> >>>> mistake anyway. So please do the right thing and make this e-mail go 
> >>>> 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 
> >> may contain information that is legally privileged, confidential and 
> >> 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 
> >> mistake anyway. So please do the right thing and make this e-mail go 
> >> 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 
> the use of the individual or entity to which it is addressed and may 
> contain information that is legally privileged, confidential and 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 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 
> 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
> 
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> 
> ------------------------------
> 
> 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 
> 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 
> and an acetone/dry ice slurry.  Either way the testes crack, usually one 
> big crack end to end.  Doesn't seem to be as much of a problem with our 
> 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 
> 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 
> greatly appreciated.
> 
> Thanks,
> Renee'
> 
> 
_______________________________________________________________________________________
> 
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> 
> ------------------------------
> 
> 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
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> 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
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> ------------------------------
> 
> 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
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> 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 
> 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 
> would handle cases like these separately. They were processed, embedded, cut 
> and stained separately. Once the case was completed, we changed all the 
> 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. 
> > 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
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> 
> ------------------------------
> 
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> 
> End of Histonet Digest, Vol 46, Issue 6
> ***************************************
------- End of Original Message -------


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