[Histonet] Re: Histonet Digest, Vol 99, Issue 2

Madeleine Huey madeleinehuey <@t> gmail.com
Sun Feb 5 00:27:06 CST 2012


Gudrun,

I don't understand why you say "IF on frozen unfixed tissue is the
usual way found in literature".

IF can be done on any type of tissues.

By any chance your doctors ask for multiple staining with the IF?
Maybe he's looking for co-localization, that's the best way.

Here's an example of IF;
1) Dep. & hydration
2) HIER; AR with Diva (Biocare) for 3'-5' with pressure cooker, rinse
with washing buffer immediately (no need cooling for 20 min @ RT)
3) 1st Ab; incubate Rabbit Monoclonal anti - C4d 1:50 (Spring
Bioscience) for 60 min in dark "you can use your existing 1st ab, but
do different titration to optimized"
4) 2nd Ab; incubate Alexa 594/___ anti-Rabbit (1:100) for 60 min in
dark "whatever 2nd Abs & titre you have in your lab"
5) Counterstain with DAPI (whatever you have in the lab) for 5 min in dark
6) Coverslip with your usual IF mounting solution (ie. Prolong gold, etc)

Madeleine Huey, HTL (ASCP) QIHC
Supervisor - Pathology (IPOX & Histology)
madeleine_h <@t> elcaminohospital.org







On Thu, Feb 2, 2012 at 10:00 AM,
<histonet-request <@t> lists.utsouthwestern.edu> wrote:
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> Today's Topics:
>
>   1. Re: Looking for Part-Time Volunteer Work (Paula)
>   2. CLIA Inspection (Fawn Bomar)
>   3. Job Listing Houston,Tx (Stella Mireles)
>   4. On call Position in Chicago (Lester Raff MD)
>   5. RE: RAC Medicare Audits - PAs (Richard Cartun)
>   6. C4d IF on FFPE kidney (Gudrun Lang)
>   7. Re: Bleaching in the histo lab (Lee & Peggy Wenk)
>   8. Re: RAC Medicare Audits - PAs (Kim Donadio)
>   9. Re: Bleaching in the histo lab (Kim Donadio)
>  10. RE: IHCRG where are you?  (JMyers1 <@t> aol.com)
>  11. AW: [Histonet] C4d IF on FFPE kidney (Gudrun Lang)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Wed, 1 Feb 2012 10:03:20 -0800 (PST)
> From: Paula <araniqkslvr <@t> yahoo.com>
> Subject: [Histonet] Re: Looking for Part-Time Volunteer Work
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID:
>        <1328119400.2279.YahooMailClassic <@t> web30302.mail.mud.yahoo.com>
> Content-Type: text/plain; charset=us-ascii
>
> I am in Raleigh, NC. Looking for part-time volunteer work to build skills up.
>
> Paula
>
>
>
> ------------------------------
>
> Message: 2
> Date: Wed, 1 Feb 2012 18:46:04 +0000
> From: Fawn Bomar <Fawn.Bomar <@t> HalifaxRegional.com>
> Subject: [Histonet] CLIA Inspection
> To: "histonet <@t> lists.utsouthwestern.edu"
>        <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>        <0111BC10D77DC54EAB99B2DDA3BCE4B916B41A <@t> EXCH-2K10.hrhs.com>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Hi everyone!
>
>
>
> I was wondering if anyone out there that has knowledge of CLIA regs and inspections would be willing to contact me directly to help me answer a few questions.
>
>
>
> My contact info is Fawn.Bomar <@t> halifaxregional.com<mailto:Fawn.Bomar <@t> halifaxregional.com> or call if it is easier at 434-517-3033.
>
>
>
> Thank you in advance,
>
> Fawn
>
>
> ------------------------------
>
> Message: 3
> Date: Wed, 1 Feb 2012 13:24:48 -0600
> From: Stella Mireles <estellamireles <@t> gmail.com>
> Subject: [Histonet] Job Listing Houston,Tx
> To: "histonet <@t> lists.utsouthwestern.edu"
>        <Histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>        <CABm4S9PmKmUWGe3MmOWMDLRRNNiYQfFGrLRbo_GFCSq4FoFHVg <@t> mail.gmail.com>
> Content-Type: text/plain; charset=windows-1252
>
> ------------------------------
>  *ANNOUNCEMENT NUMBER: 14905- T*
> * *
> *JOB TITLE:               Histology Technician II *
> * *
> *DEPARTMENT:        Institute of Forensic Sciences*
> * *
> *HOURS:                     7:30 a.m. – 4:40 p.m. / Flexible*
> *                                    Monday - Friday*
> * *
> *SALARY:                    Commensurate With Experience*
> *                                    Based On 26 Pay Periods*
> * *
> *EDUCATION:  *Completion of an Associate’s degree and completion of
> histology school with histo-technician certification (ASCP) American
> Society of Clinical Pathologist *at the time of employment or within one
> year of employment.  *
> * *
> *EXPERIENCE:  *One year of experience in a histology laboratory in which
> responsibilities included production of stained slides and preservation of
> tissue samples in both paraffin and formalin is *required*.**
> * *
> *JOB SKILLS:  *The successful applicant must have expertise in the use of
> microtomes, manual staining procedures, manual slide coverslipping,
> automated slide stainers, automated slide coverslipper, tissue processors
> and tissue embedding.  Must be capable of understanding and adhering to
> strict protocols for the handling, trimming and archival of tissue samples
> and blocks; knowledge of histology laboratory safety rules and procedures *is
> essential*.  Good interpersonal skills *are a must*.
>
> *JOB DESCRIPTION:  *Prepares stained slides of autopsy tissues in a
> careful, controlled environment.  Prepares paraffin blocks of tissue for
> long-term storage and labels the samples in accordance with histology
> laboratory protocols; assists with archival of paraffin-embedded and
> non-paraffin-embedded formalin fixed tissues. Provides all tissue slides to
> the assigned Assistant Medical Examiner, Deputy Chief Medical Examiner or
> Chief Medical Examiner on a timely basis; assists with data entry into a
> computerized database to track laboratory efficiency as required.  Other
> job assignments as assigned by the Deputy Chief Medical Examiner.  *Position
> requires a high level of confidentiality, responsibility and dependability.*
>
> *PHYSICAL REQUIREMENTS:**  *Must be able to sit and stand for prolonged
> periods of time; able to lift up to 40lbs; stooping and bending may be
> required.
> * *
> *EMPLOYMENT IS CONTINGENT UPON PASSING A CRIMINAL BACKGROUND CHECK.*
> * *
> * *
> *HARRIS COUNTY HAS AN EMPLOYMENT AT WILL POLICY.*
> *CLOSING DATE:      Open Until Filled*
> *APPLY AT:                1310 PRAIRIE - SUITE 170*
> * *
> * *
> *UPON RECEIVING A CONDITIONAL OFFER OF EMPLOYMENT, ALL APPLICANTS ARE
> SCREENED FOR THE PRESENCE OF ILLEGAL DRUGS.***
>
>
> ------------------------------
>
> Message: 4
> Date: Wed, 1 Feb 2012 13:37:55 -0600
> From: "Lester Raff MD" <LRaff <@t> uropartners.com>
> Subject: [Histonet] On call Position in Chicago
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID:
>        <FE0368FE5AA6684DB5990BBEED742C53010BDC07 <@t> UPHQMSX01.uropartners.local>
> Content-Type: text/plain;       charset="us-ascii"
>
> Our private outpatient specialty lab has an opening for an on-call
> part-time second shift and weekend histologist. We are located in the
> western Chicago suburbs about a mile east of Oak Brook Shopping Center.
> If interested, please contact Andrea O'Brien at 708-486-0076
> <about:blank> . Experienced histologists only, please.
>
>
>
>
>
>
>
> Lester J. Raff, MD
>
> Medical Director
>
> UroPartners Laboratory
>
> 2225 Enterprise Dr. Suite 2511
>
> Westchester, Il 60154
>
> Tel 708.486.0076
>
> Fax 708.492.0203
>
>
>
>
>
> ------------------------------
>
> Message: 5
> Date: Wed, 01 Feb 2012 14:45:06 -0500
> From: "Richard Cartun" <Rcartun <@t> harthosp.org>
> Subject: RE: [Histonet] RAC Medicare Audits - PAs
> To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>,     "Loralee A
>        McMahon" <Loralee_Mcmahon <@t> URMC.Rochester.edu>
> Message-ID: <4F294FF2.7400.0077.1 <@t> harthosp.org>
> Content-Type: text/plain; charset=US-ASCII
>
> I don't know.  I was hoping that there is someone in the Histonet community that is familiar with this.
>
> Richard
>
>>>> "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu> 2/1/2012 8:56 AM >>>
> I haven't heard that one.   Do they mean present physically or present as in their office reading slides, but a phone call or page away?
>
> Loralee McMahon, HTL (ASCP)
> Immunohistochemistry Supervisor
> Strong Memorial Hospital
> Department of Surgical Pathology
> (585) 275-7210
> ________________________________________
> From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Richard Cartun [Rcartun <@t> harthosp.org]
> Sent: Tuesday, January 31, 2012 7:19 PM
> To: Histonet
> Subject: [Histonet] RAC Medicare Audits - PAs
>
> Is anyone familiar with the new requirement effective January 1st, 2012 that states that Pathologists' Assistants can no longer teach residents unless a pathologist is present?
>
> Richard
>
> Richard W. Cartun, MS, PhD
> Director, Histology & Immunopathology
> Director, Biospecimen Collection Programs
> Assistant Director, Anatomic Pathology
> Hartford Hospital
> 80 Seymour Street
> Hartford, CT  06102
> (860) 545-1596 Office
> (860) 545-2204 Fax
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 6
> Date: Wed, 1 Feb 2012 21:17:14 +0100
> From: "Gudrun Lang" <gu.lang <@t> gmx.at>
> Subject: [Histonet] C4d IF on FFPE kidney
> To: <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <138B99031E744674A4C020CB44691D27 <@t> dielangs.at>
> Content-Type: text/plain;       charset="us-ascii"
>
> Hi!
>
> Can someone provide me a immunofluorescence protocol for C4d on formalin
> fixed human paraffin sections?
>
>
>
> thanks in advance
>
>
>
> Gudrun Lang
>
>
>
> ------------------------------
>
> Message: 7
> Date: Wed, 1 Feb 2012 18:03:21 -0500
> From: "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net>
> Subject: Re: [Histonet] Bleaching in the histo lab
> To: "angela smith" <we3smitty <@t> yahoo.com>,
>        <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <A4060F6BA3CA447B99D81738529CCF0F <@t> HP2010>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
>        reply-type=original
>
> Several questions and comments, in no particular order:
>
> 1. What percent of bleach?
> - 10% is all that is needed for biohazards. If you are concerned about the
> smell, it might be too high a percent.
>
> 2. How good is your ventilation? How long do you continue to smell the
> chlorine?
> - If you continue to smell it hours later, or even the next day, have your
> safety officer and maintenance people check out the ventilation.
>
> 3. After wiping down with 10% bleach, are you wiping down the counter with
> water?
> - Need to clean off the corrosive bleach off the surfaces. That would also
> help with the smell. But takes more time.
>
> 3. What locations in the lab are you cleaning with dilute bleach?
> - The only areas that need to be cleaned with a disinfectant are those areas
> that have fresh or not completely fixed tissue, so around the grossing
> stations and the cryostat. Maybe where specimens are received into the lab.
> - The areas where you process tissue, embed, microtome, do staining, file
> slides and blocks should not need to be disinfected with bleach. The tissue
> has been fixed in formalin, and gone through alcohol, xylene (or
> substitute), and placed in 60 degree C (140 degree F) paraffin. That should
> kill almost all microorganisms. Therefore, should not need to clean up with
> anything beyond soap and water. If you have a very underprocessed tissue
> block, and it's oozing and weeping all over the counter and microtome, you
> may want to disinfect the area. (If it's a CJD case, you are going to need
> strong solutions than 10% bleach, but that's a whole new conversation.)
> - So talk with your safety officer, about how there are no biohazards in the
> other parts of the lab. They may be thinking more of the clinical pathology
> labs, with blood tubes and petri dishes, needing to be disinfected with
> bleach every day/shift.
>
> 4. Chemical incompatibility:
> Bleach is incompatible with ammonia (makes chlorine gas - deadly)
> Bleach is incompatible with acids
> Bleach is an oxidizer, and formaldehyde is supposed to be kept away from
> oxidizers.
> So, yes, I would be a little worried about chemical interaction. However,
> wiping down the area first with water, to remove other chemicals, before the
> bleach, would take care of this problems.
>
> 5. What does Epidemiology suggest for disinfectant?
> Our epidemiology is suggesting other cleaning solutions for disinfecting,
> rather than bleach, in many cases.
> - not as corrosive
> - less obnoxious fumes
> - more "green"
> - better disinfectant and faster, than bleach
>
> Peggy Wenk, HTL(ASCP)SLS
> Beaumont Health Systems
> Royal Oak, MI 48073
> (Comments reflect my opinions, not that of my hospital)
>
> -----Original Message-----
> From: angela smith
> Sent: Wednesday, February 01, 2012 8:57 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Bleaching in the histo lab
>
> I have been told by our safety officer that it is standard practice too
> clean the lab at the end of the day with diluted bleach. I have noticed a
> chemical reaction (smell) when cleaning the main area of the lab. I have
> concerns that this is not a good practice due to chemical reactions as we
> use so many chemicals in histology. What do other people do?  Also I believe
> it is unsafe to use bleach with anything formalin related.
> Please let me know if you have a "standard" practice or mandated cleaning
> from your facility.
> Angela
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 8
> Date: Wed, 1 Feb 2012 19:26:45 -0500
> From: Kim Donadio <one_angel_secret <@t> yahoo.com>
> Subject: Re: [Histonet] RAC Medicare Audits - PAs
> To: Richard Cartun <Rcartun <@t> harthosp.org>
> Cc: Histonet <histonet <@t> lists.utsouthwestern.edu>,       Loralee A McMahon
>        <Loralee_Mcmahon <@t> URMC.Rochester.edu>
> Message-ID: <65FAEDF0-7CFC-4188-8061-2382A42A7C00 <@t> yahoo.com>
> Content-Type: text/plain;       charset=us-ascii
>
> Sorry.  But I thought RAC was a group of individuals who go over your billing looking for over payments to Medicare   They typically get a % of what they find so it motivates them. If u have this group of auditors in your area and they Are saying this then I would just ask them to show you the rule they must be referring to a CLIA guideline somewhere.
> Best of luck
> Kim D
>
> Sent from my iPhone
>
> On Feb 1, 2012, at 2:45 PM, "Richard Cartun" <Rcartun <@t> harthosp.org> wrote:
>
>> I don't know.  I was hoping that there is someone in the Histonet community that is familiar with this.
>>
>> Richard
>>
>>>>> "McMahon, Loralee A" <Loralee_Mcmahon <@t> URMC.Rochester.edu> 2/1/2012 8:56 AM >>>
>> I haven't heard that one.   Do they mean present physically or present as in their office reading slides, but a phone call or page away?
>>
>> Loralee McMahon, HTL (ASCP)
>> Immunohistochemistry Supervisor
>> Strong Memorial Hospital
>> Department of Surgical Pathology
>> (585) 275-7210
>> ________________________________________
>> From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Richard Cartun [Rcartun <@t> harthosp.org]
>> Sent: Tuesday, January 31, 2012 7:19 PM
>> To: Histonet
>> Subject: [Histonet] RAC Medicare Audits - PAs
>>
>> Is anyone familiar with the new requirement effective January 1st, 2012 that states that Pathologists' Assistants can no longer teach residents unless a pathologist is present?
>>
>> Richard
>>
>> Richard W. Cartun, MS, PhD
>> Director, Histology & Immunopathology
>> Director, Biospecimen Collection Programs
>> Assistant Director, Anatomic Pathology
>> Hartford Hospital
>> 80 Seymour Street
>> Hartford, CT  06102
>> (860) 545-1596 Office
>> (860) 545-2204 Fax
>>
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet <@t> lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 9
> Date: Wed, 1 Feb 2012 19:51:45 -0500
> From: Kim Donadio <one_angel_secret <@t> yahoo.com>
> Subject: Re: [Histonet] Bleaching in the histo lab
> To: Lee & Peggy Wenk <lpwenk <@t> sbcglobal.net>
> Cc: "<histonet <@t> lists.utsouthwestern.edu>"
>        <histonet <@t> lists.utsouthwestern.edu>
> Message-ID: <E3306E2A-EEB7-4E87-9651-6B7B6BD5CA7F <@t> yahoo.com>
> Content-Type: text/plain;       charset=us-ascii
>
> Maybe those scented bleach wiped would work. I love those things and they smell pretty good.  Just a thought. I like the greencan fresh scent
> products/clorox-disinfecting-wipes/
> KimD
> Sent from my iPhone
>
> On Feb 1, 2012, at 6:03 PM, "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net> wrote:
>
>>
>> - Need to clean off the corrosive bleach off the surfaces. That would also
>
>
>
> ------------------------------
>
> Message: 10
> Date: Thu, 2 Feb 2012 06:38:33 -0500 (EST)
> From: JMyers1 <@t> aol.com
> Subject: RE: [Histonet] IHCRG where are you?
> To: histonet <@t> lists.utsouthwestern.edu
> Cc: broyce <@t> nsh.org, cdiamond <@t> nsh.org, Carol.Freeman <@t> utoledo.edu
> Message-ID: <9a4e.313de094.3c5bcfb9 <@t> aol.com>
> Content-Type: text/plain; charset="US-ASCII"
>
> Carol:
> I'm happy to report that the IHC Resource Group remains alive and  well.
> For reasons unkown, the group's website is currently not working  properly,
> and through this message I've notified members of the NSH's staff  of the
> problem.  We are actively working on a number of projects, most  notably the
> annual IHC Forum event, and once our site is working again we hope  to post new
> information.  Stay tuned for details...
> Sincerely,
> Joe Myers, M.S, CT(ASCP)QIHC
> Chair, IHC Resource Group
> ------------------------------
>
> Message: 9
> Date: Wed, 1 Feb 2012  10:33:08 -0500
> From: "Freeman, Carol"  <Carol.Freeman <@t> utoledo.edu>
> Subject: [Histonet] IHCRG where are  you?
> To: _histonet <@t> lists.utsouthwestern.edu_
> (mailto:histonet <@t> lists.utsouthwestern.edu)
>
> Good  Morning,
>
> Just curious if any one out there knows what happened to the  IHCRG (IHC
> review group?) It was an organization advertised in the NSH  material I
> brought home from last September but the website doesn't exist and  the
> email address listed was undeliverable...Just curious It sounded like  an
> interest idea, just wondered what happened.. Thank you for any  response
> :)
>
>
> ------------------------------
>
> Message: 11
> Date: Thu, 2 Feb 2012 15:31:04 +0100
> From: "Gudrun Lang" <gu.lang <@t> gmx.at>
> Subject: AW: [Histonet] C4d IF on FFPE kidney
> To: "'Richard Cartun'" <Rcartun <@t> harthosp.org>
> Cc: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <206DA8F070EC4221B7C45F02B3A99C1C <@t> dielangs.at>
> Content-Type: text/plain;       charset="iso-8859-1"
>
> We have been performing C4d IHC on FFPE kidney for a couple of years. We
> receive only fixed samples.
> Yesterday my doctors came with this idea of IF - I still have to ask for the
> special reasons. But I have the suspicion, that they are not aware of the
> fact, that IF on frozen unfixed tissue is the usual way found in literature.
>
> There are some articles that deal with comparison of IF(frozen) and
> IHC(ffpe). The results are usually an equal outcome. IF(frozen) shows
> additional staining in glomeruli.
>
> Perhaps someone told them, that IF is the golden standard and recommended.
>
> Gudrun
>
> -----Ursprüngliche Nachricht-----
> Von: Richard Cartun [mailto:Rcartun <@t> harthosp.org]
> Gesendet: Mittwoch, 01. Februar 2012 21:21
> An: gu.lang <@t> gmx.at
> Betreff: Re: [Histonet] C4d IF on FFPE kidney
>
> Hi Gudrun:
>
> Is there reason why you want to use IF and not immunoperoxidase?
>
> Richard
>
> Richard W. Cartun, MS, PhD
> Director, Histology & Immunopathology
> Director, Biospecimen Collection Programs
> Assistant Director, Anatomic Pathology
> Hartford Hospital
> 80 Seymour Street
> Hartford, CT  06102
> (860) 545-1596 Office
> (860) 545-2204 Fax
>
>
>>>> "Gudrun Lang" <gu.lang <@t> gmx.at> 2/1/2012 3:17 PM >>>
> Hi!
>
> Can someone provide me a immunofluorescence protocol for C4d on formalin
> fixed human paraffin sections?
>
>
>
> thanks in advance
>
>
>
> Gudrun Lang
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> End of Histonet Digest, Vol 99, Issue 2
> ***************************************



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