Re: [Histonet] Fast Green / Sirius Red - Unknown blue features

David We have found that mordanting in bouins (just like you would a Masson's trichrome) prior to completing a standard Picrosirius Red stain protocol that we get very consistent results. I think this method has been published but I do not have access to the reference currently. Its worth a

Re: [Histonet] Desperately seeking a certain IHC expert

Joe Myers biocate Sent from my Windows 10 device From: Maria Cruz via Histonet Sent: Monday, February 3, 2020 7:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Desperately seeking a certain IHC

Re: [Histonet] IHC on Pig Tissues

Lori You can't use a dual label detection system. We have found most mouse polymers will cross react with porcine tissue. We know that the dako one does. You need to use the rabbit envision for rabbit monoclonal antibodies or with mouse monoclonals you need to use a rabbit anti-mouse and

Re: [Histonet] H trouble shooting

That an image of incomplete dehydration the samples have not been processed properly and adequately dehydrated. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell

Re: [Histonet] guidelines for using a microwave designated for food, but used to heat up histogel

You don’t need to heat up histogel in a microwave its quicker that way but if you set an oven at around 65 to 70 it will melt in around 20 minutes or so, that’s what we do. It can get messy in a microwave. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592

Re: [Histonet] Roche Symphony and Ultra users

It might be the coverglass we have seen bubbles in the glass itself, it looks like there is water on the slide, but it's actually bubbles in the coverglass Sent from my Windows 10 device From: Victoria Baker via Histonet Sent: Friday, August 30, 2019

Re: [Histonet] Antibody Validation CLIA

Keep in mind like Lacy said its cases and not slides so you could place multiple cases on one slide or create a simple array using disposable biopsy punches. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060

Re: [Histonet] Image analysis on breast markers

Teresa CAP should be releasing a paper soon, within the next couple months. Quantitative Image Analysis (QIA) for HER2 Immunohistochemistry for Breast Cancer, it will address the question that you have. Keep any eye out for it. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier

Re: [Histonet] Mouse bladder

I would try histogel instead of agar. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com

Re: [Histonet] Beaker with or without Vantage

I'm a member of API - Association for Pathology Informatics and they have a list server similar to the histonet and there is always posts about Epic Beaker. You might find some information on their website. https://www.pathologyinformatics.org/ Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC

Re: [Histonet] Tracking lot numbers

Teri When we change the tissue processor or H stainer instead of just recording a "check mark" that the solution has been changed we record the lot numbers on the form instead. We also have a from that is posted above the tissue processor that we fill in with dry erase the current lot numbers

Re: [Histonet] Old control blocks

Elaine We have the temperature of our cryostat calibrated yearly by an outside calibration company. If the thermometer you are using inside the cryostat has been calibrated and is traceable I think you might have an issue with the temperature reading that is being displayed on your cryostat.

RE: [Histonet] Paraffin processing and embeding of Chondrocyte pellets.

Bret We have done a bit of work with these types of samples, the first thing we do is remove the formalin and add eosin to the container for about 5 minutes. Then we place the pellet (using a plastic disposable pipettor) in a mesh bag and process on our tissue processor on a cycle that is set at

RE: [Histonet] (no subject)

Dorothy I would assume that you would just run the validation as you would any other antibody that is not a pre dilute. Run the antibody protocol on the number of tissues that you normally do to validate, evaluate and record the results. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager

[Histonet] RE: [IHCRG] laminin and collagen4 in cat and dog

Margaret We have used the Collagen IV listed below on canine but not feline, for laminin we have not used the antibody from abcam on canine or feline but the spec sheet states it works on most mammals. Good Luck Liz Collagen IV SouternBio 0340-01 UNLB Goat Laminin abcam

RE: [Histonet] Fume hood

We have a service come out and inspect our hoods once a year. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont,

RE: [Histonet] IHC pos. neg. control question

Amos Isotype negative controls are based upon protein concentration not dilution. They must be the same protein concentration of the primary antibody at the dilution you are using. Using the same dilution will not work unless both stock solutions (primary antibody and isotype control) are at

RE: [Histonet] IHC pos. neg. control question

, Colorado 80504 -Original Message- From: Amos Brooks [mailto:amosbro...@gmail.com] Sent: Friday, May 20, 2011 3:05 PM To: Liz Chlipala Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IHC pos. neg. control question Thanks Liz, You are absolutly right there, but have you

RE: [Histonet] Looking for Aperio or similar Slide Scanner

Kathy We have an Aperio Scanscope XT and scan for a fee Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont,

RE: [Histonet] sections falling off slides

Robin We run quite a few IHC's on goat bone femurs. We use a very good plus slide. Dry flat at 40C overnight and let them sit a few days before we stain, we do not place them in a 60C oven since we find that the articular cartilage may flip. We have good success with proteinase K with staining

RE: [Histonet] preference: fat stains

Michelle We use osmium all of the time, we find it works better for image analysis than the Oil Red O. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to

RE: [Histonet] TNF alpha antibody

We have used the antibody from abcam on FFPE mouse tissue with good success, have not tired it on frozen sections Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com

RE: [Histonet] decalcification of articular cartilage

We do a lot of IHC staining on bone and cartilage samples and we use 10% formic acid for decalcification for immunohistochemistry. I'm afraid you may not be able to turnaround this in a week. The tissue needs to be adequately fixed prior to decalcification, and properly decaled. I would trim

RE: [Histonet] processing times for skin specimens

Carol We are a research lab but we have always processed our skin samples on longer processing cycles, even the punch biopsy samples. The shaves might be different, if I wanted to cut down I would start with how you normally process them and then cut down at 5 minute increments to start and see

RE: [Histonet] Sad news from Nebraska

That has to be an April Fool joke, got ya!!! Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504

RE: [Histonet] HercepTest

We use the PT Link unit for Hercept Test Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504

RE: [Histonet] Verhoeff/Masson's Stain

John We do this stain all of the time, but we make up our reagents from scratch we don't use a kit. My suggestion is to make up your elastic stain fresh. We make up our elastic stain fresh and use it only once (or for that day) and then discard. I have attached our procedure in a different

RE: [Histonet] RE: comprehensive list?

Vendor sheets may not be accurate we keep an internal log of all of our antibodies and what species we have tested them in. So far in our lab we have found several antibodies that the vendor states works in a species and it does not in our hands, in these cases we always contact the vendor,

RE: [Histonet] Processing animal fat

5mm is pretty thick still. We process skin and fat and we use a longer processing cycle. 1 to 1.5 hours per station. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com

RE: [Histonet] Processing animal fat

The laser treatment should not be an issue, we do that type of work all of the time. I would trim to about 2-3 mm. I did notice that you only have 2 changes of citrisolv a xylene substitute I have not worked with that one in particular but if I do work with a xylene substitute we use three

RE: [Histonet] Research and Clinical Labs

Amos We are a GLP compliant contract research lab, and we are currently in the process of applying for a CLIA accreditation. I don't see why you would need to have separate equipment, etc. In our case we will not be processing standard diagnostic samples but we feel that the CLIA accreditation

RE: [Histonet] VALIDATION of a new clone

If it's a different clone then it's essentially a different antibody you would have to go through the same process as you did initially. LIz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060

RE: [Histonet] Her2 Gastric cases

Margaret Dako has a bunch of info on Her2 staining in gastric cancer its in the most recent Connection Volume 15 and they also have a companion document that covers Her 2 staining and scoring for gastric cancer, just go to the Dako website and you should be able to view these documents. Liz

RE: [Histonet] Competancies for handling hazardous material

) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala Sent: Friday, March 04, 2011

RE: [Histonet] Mouse cardiac myosin antibodies

Mark I took a brief look at the abcam antibody and in particular the review with the paraffin staining. If I was purchasing the antibody I would question the results of that review, since they used a goat anti-mouse detection system which is not ideal for mouse tissue. Plus I'm not sure that

RE: [Histonet] Polymer detection kit on rat tissue

Anita Since you are working on rat tissue, you have to be careful on what detection systems you use. You will need two different detection systems for your work. One for the mouse monoclonal antibodies and one for the rabbit polyclonal. If you want to use a polymer based detection system

RE: [Histonet] Digital Pathology/Telepathology

Andrew We are a research contract lab and have been scanning slides (WSI) since 2007. We have an Aperio Scanscope XT. Our clients have secure remote access to their images. We manage our own server, storage (20TB) and backup locally. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager

RE: [Histonet] Competancies for handling hazardous material

Rae We have, you need standard training, but the individuals who inspected our lab did not mention compentances. But our inspection happened because we needed to switch from a conditionally exempt small quantity generator to a small quantitiy generator and we then needed to register with the

RE: SPAM-LOW: [Histonet] TRAP staining kit

I'm in agreement with Patsy. We have found in our hands that the Sigma Kit will not work on formic acid decaled bone. It may work on EDTA decaled bone, the reason I say may is that we have gotten variable staining results with EDTA decaled bone. For mouse knees and ankles we have gotten

[Histonet] IgG and IgM on mouse kidneys

Hello all I have a quick question. Can I perform IgG and IgM staining on mouse kidneys on FFPE tissue samples? I thought that this type of staining is normally completed on frozen sections via Immunofluorescence. Any advice would be helpful Thanks Liz Elizabeth A. Chlipala, BS,

RE: [Histonet] Information Needed

Pamela We use a hepa filtered vacuum from Marmed, Inc. (440) 572-5175 LiZ Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E

RE: [Histonet] processing histogel

Luciana You process histogel samples essentially the same way you process regular tissue samples. I'm assuming you will be dealing with a about a 1 cm cube sample that you want embedded into the histogel and then you would then slice the histogel cube (with the collagen construct) into 3 mm

RE: [Histonet] bar-coded chemical inventory

I would be interested in that info too. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504

RE: [Histonet] Help with OCT problem

Donna I have seen soft or rather sticky OCT samples in the past. If you freeze in isopentane and don't let the isopentane evaporate off the samples prior to wrapping the sample in foil, that excess isopentane changes the OCT, it makes it sticky. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC

RE: [Histonet] IHC validation

Joe If you are following the recommendations from the CAP paper on IHC standardization then it would be 25 tissues (10 strong positive, 10 weak to moderate positive and 5 negative). Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado

RE: [Histonet] IHC validation

Chlipala; Joe Nocito; Histonet Subject: RE: [Histonet] IHC validation But that is for receptors, correct? Do you do that for everything? Thanks, j -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala

RE: [Histonet] IHC validation

...@ucsfmedctr.org] Sent: Tuesday, February 08, 2011 4:38 PM To: Liz Chlipala; Weems, Joyce; Joe Nocito; Histonet Subject: RE: [Histonet] IHC validation When changing instruments you are validating the instrument, not the test. For each antibody you just need to run parallel tests in each instrument showing

RE: [Histonet] Rat lung histology for mast cells and eosinophils

Chen For mast cells you can fix in 10% NBF and be fine with toluidine blue. We stain with toluidine blue on formalin fixed samples all of the time. The trick is not to dehydrate the sections, let them air dry and then mount. I would stick with formalin fixation that's going to be your best bet

RE: [Histonet] Her2 Fixation Requirement

Lori I would think both if you could since the Her2 paper does address those issues as potentials for problems in testing variation (time to fixation). I believe the CAP/ASCO paper states not less then 6 and no more than 48 hours for fixation (2007 paper). I believe you also should document

[Histonet] EGF IHC

Hello everyone Does anyone out there have a good EGF (not EGFR) antibody that works well in formalin fixed paraffin embedded samples - species is human Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308

[Histonet] slide labeler advice

Hello again We are in the market for a slide labeler. We currently have the SlideMate from Thermo Scientific. We have had it for about 2 years now and we are just not happy with the quality and consistency of the printing. We are a contract research lab and not a clinical lab. Entering data

RE: [Histonet] stains for visualizing new bone growth

Robin I'm not sure how accurate this is, but we do a lot of massons trichrome staining in rat muscle pouch studies (they inject into the muscle pouch something that will induce bone formation) and we have noticed that new bone formation has a tendency to stain blue rather than red. You could

RE: [Histonet] ventilation and exposure

Curt We fill our bottles in the hood. They are emptied into a vented flammable cabinet that contains a 55 gallon drum. You know you can do your own chemical badges for both xylene and formalin. I like to run a couple TWA (8 hour) exposures and then I badge a lot of STEL (15 minutes) exposures

RE: [Histonet] Cytoplasmic staining with Ki67 in frozen sections

James I believe the nuclear staining pattern that you describe is normal for Ki-67 in the spleen, not sure about the cytoplasmic or particulate staining. Since you viewing with fluorescence the particulate staining may be autofluorescence of red blood cells. Normally our primary antibody

RE: [Histonet] Oil Red O for FS on Muscle

Allison We make up our Oil Red O from scratch same day we use it, let it stand 10 minutes, and then filter with a Millipore Stericup 0.22µm, GP Express Plus Membrane, 250ml Receiver Bottle, catalog number: SCGPU02RE under vacuum. That seems to help a bit with the background and we are working

RE: [Histonet] Consult Fees for IHC

Joe Keep in mind that that what you charge you need to take into consideration the taxes you will be responsible for paying. Most consultants are paid a simple fee and the company paying you will not take out any taxes. So you need to account for that. I have known quite a few individuals who

RE: [Histonet] coverglass

That's who we use and we don't have any problems, with have the Tissue Tek Glas coverslipper Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567

[Histonet] Feulgen Stain for DNA

Hello Everyone I have tried to find this answer on the web and in references but I need some help. With respects to the Feulgen stain, how sensitive is it? Can it detect smaller fragments of DNA? Does it have to be double stranded or will single stranded stain as well? Thanks in advance.

RE: [Histonet] Research charges - long reply

Mary Mary We are a private contract lab and do not charge for recuts or repeats that are related to quality issues. We run a lot of bone samples through our lab and if there is a fold in the lesion area we recut the block at no charge to the client. All of our slides are QC'd and we will

RE: [Histonet] Lab chairs

Sharon We got chairs from VWR - catalog number 80086-440 VWR(r) Contour(tm) Self-Skinned Urethane Chairs, we got these back in 2007 or 2008 and they have held up great, they are comfortable and clean very easily. They might be a bit pricey. I think I paid around $150.00 to $200.00 for each back

[Histonet] FW: ASCP Action Alert - Physician Signature Requirement _FYI

FYI - saw alot about this today, just got this e-mail from ASCP Liz -Original Message- From: American Society for Clinical Pathology [mailto:a...@site1.ascpmail.org] Sent: Wed 12/22/2010 2:42 PM To: Liz Chlipala Subject: ASCP Action Alert - Physician Signature Requirement http

RE: [Histonet] Reg: Beta-2 minroglobin IHC

Robin Have you tried other retrieval methods, its best to normally try to stay away from HIER on bone but sometimes you have to use it. If you have not already tried other retrieval methods I would try no retrieval, proteinase K and then trypsin or pronase, my favorite is proteinase K. Since the

RE: [Histonet] Decalcification

Try surface decaling the samples in the block. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504

RE: [Histonet] neurodegeneration stains?

Anjum Those are the two that I'm familiar with. I'm not aware of any others. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive,

RE: [Histonet] Cheap slide scanner for rat brains?

Caroline The older models of the Nikon scanner for kodachromes had an attachment that would scan glass slides, I'm not sure about the new ones but we had one back in the late 90's early 2000's that would hold a slide and we could scan that. You could also take some low mag images on a

RE: [Histonet] Recommendations for decal solution

I like 5 to 10% formic acid Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original

RE: [Histonet] Amputation transport

Try mopec -Original Message- From: Fleming, Jackie M jackie.flem...@allina.com Sent: Wednesday, October 13, 2010 7:59 AM To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Subject: [Histonet] Amputation transport Does anyone have a transport/ carrier for

[Histonet] ECM stain for growing cartilage

Hello Everyone Is anyone out there aware of an ECM stain that can be used on growing tissue culture cells in a hydrogel. What I'm really looking for is a live stain I can use on the whole gel and see with a light microscope. I want to get a general idea of the amount of ECM that is growing.

RE: [Histonet] Bone IHC

We lower the temp of retrieval to 70C for 2 hours and have good success with that. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway

RE: [Histonet] TUNEL

We use the Roche Kit with good success. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504

RE: [Histonet] Collagen Type II antibody

Joel We use an antibody we purchase from MD Biosciences - its quite expensive but we find it works really well for us on multiple species with chondroitinase digestion. Protocol: Rabbit anti Human Collagen Type II Clone: Polyclonal Vendor: MD Biosciences Catalog Number: MD20211 Species: Rabbit

RE: [Histonet] FW: anti-Ly-6G (aka GR-1 ) IHC question

Brett We have not used that antibody but we have used the following antibody from Serotec with good success. Protocol: Mouse Neutrophil Immunohistochemical Stain Clone: 7/4 Vendor: Serotec Catalog Number: MCA771 Species: Mouse Isotype: Rat IgG2a Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC

RE: [Histonet] Procollagen

Mary We use the one from Abcam ab64409, it works really nice Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont,

RE: [Histonet] Canine bone sectioning trouble

Overall your fixation, decalcification and processing cycle is too short. The sample size is really big, why do you need it so thick? I would first of all let the samples fix longer 24 to 48 hours or I would trim them so that they are about 3-5 mm in thickness. Either way I like fixing bone

RE: [Histonet] cryojane question

Emily You can put more than one section on a slide if you need to, but in our experience it does not work as nicely as depicted all of the time, it can be a bit tricky to work with on undecalcifed bone and harder tissues. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory,

RE: [Histonet] ISH on histogel cell pellets

Kim the histogel should not interfere with the IHC staining, I'm not sure about the ISH Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway

RE: [Histonet] how to prevent foldings on femoral head cartilage tissue

Sometimes placing them on a hot plate at about 60C will help get out the folds, the paraffin needs to melt and the sections need to turn clear then take it off the hot plate. If you leave it too long on the hotplate the articular cartilage may fold over on itself. Liz Elizabeth A. Chlipala, BS,

RE: [Histonet] Antibody Validation - long response

Bottom line it's not the vendors responsibility to validate their equipment or antibodies in your lab. Some vendors may help you do this, but ultimately the lab needs to validate the equipment and IHC in their lab. The vendors normally calibrate the equipment prior to shipment and once they

RE: [Histonet] Mounting Medium for Immunofluorescence

I think that the propidium iodide will stain the nuclei or DNA, similar to DAPI, so this mounting media will counterstain the nuclei and its visible in the FITC wavelength (488), while if you conterstain with DAPI it is excited with a different wavelength, therefore it might interfere with the

[Histonet] RE: PBA for Immunofluorescence

80504 -Original Message- From: Nails, Felton [mailto:flna...@texaschildrens.org] Sent: Thursday, May 20, 2010 1:29 PM To: Liz Chlipala; Laurie Colbert; histonet@lists.utsouthwestern.edu Subject: PBA for Immunofluorescence Concerning Immunofluorescence, where are you getting your (PBA

RE: [Histonet] weekend fixation

I was talking to Peggy Wenk over the weekend at the MSH meeting and they had a paper that was published regarding fixation and ER/PR staining sensitivity etc. The biggest problem that they reported is underfixation is much worse than over fixation. I think a minimum of 10 hours of fixation

RE: [Histonet] IHC Validation on new instrument

We perform our entire validation process as a new piece of equipment. Our validation protocols are quite extensive, up to about 85 pages long on each piece of major equipment, at least that's what it was for our new prisma stainer and glass coverslipper. We perform an installation/operational

RE: [Histonet] PPE

I require gloves when sectioning only because it helps eliminate epithelial floaters, with respects to embedding the tech can choose to or not to use gloves. No safety goggles are required. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder,

RE: [Histonet] IHC Validation (again)

Laurie I'm not aware of a particular question, but I would believe you would have to perform some validation steps for each antibody. I would approach it the same way you approach validating new lots of antisera. The CAP paper on standardization of IHC recommends 25 different samples when you

[Histonet] Article for Er/PR vaidation

Hello everyone I have been trying to locate this article on line and I'm either challenged or its not there. Its from the Arch Pathol Lab Med (in press) it says in press so I don't know what that that means but its not available off the Journals web site Fitzgibbons, P. L. , D. A.

RE: [Histonet] Thanks for bone cutting suggestions - and anti-Rat CD3

Dako has a rabbit polyclonal that works in rat. The other thing with the bone, is place the slide with a section on a flat hotplate (60 C) until it turns clear - not too long or the articular cartilage will flip. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO

RE: [Histonet] CBG BIOTECH RECYCLER

Nirmala We have the CBG recycler also. We initially started recycling both alcohol and xylene, but we now only recycle alcohol. We ended up with so much recycled xylene that we could not use. Also you can not use the recycled xylene on the last xylene station in the tissue processor, or for

RE: [Histonet] Responses to IHC CAP Validation question

CAP does have guidelines, it's a paper that was published a few years back, its right on the CAP website. I have it if anyone needs it. I'm not sure they use it for their inspections but it covers how to validate an antibody. How many samples to run, what paperwork needs to be kept, etc. Liz

[Histonet] tape coverships

Hello all I have a question on tape coverslips. We received a couple tape coverslipped slides in for scanning and the tape is completely removed from the slide. I would normally just coverslip with glass but the tissue is on the tape coverslip and not on the slide. How can I re attach the

RE: [Histonet] tape coverships

: Mike Pence [mailto:mpe...@grhs.net] Sent: Wednesday, April 21, 2010 3:11 PM To: Liz Chlipala; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] tape coverships Just put the slide in xylene and coverslip with the tape containing the tissue. The you could put a couple small drops

RE: SPAM-LOW: Re: [Histonet] Cd31 in pigs

Patsy How do you order that rat anti-mouse CD31. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont,

RE: [Histonet] 72644.18148...@web111105.mail.gq1.yahoo.com

Thomas Just a question, what would you consider an unnecessary CAP regulation? Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive,

RE: [Histonet] Incubation of Ab more then 1 hour in autostainer

We have done up to an hour but not longer, if I were you I would have two antibody steps and just to add some more reagent after an hour or 1.5 hours Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303)

RE: [Histonet] CD3 on mouse

Dako's rabbit anti-CD3 cross reacts with mouse Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504

RE: [Histonet] tissue processing

Sheri Your processing cycle is too long. We process 20 minutes per station for mouse tissue only 2 absolutes and 2 xylenes and 3 paraffins. Even 3 absolutes and 3 xylenes at 20 minutes a station we find over processes the tissue. Liz From:

RE: [Histonet] Saffron/Eosin as a counterstain for HE???

Its hematoxylin, ploxine and saffron. Believe it or not it was the stain that we used at my first job back in 1973. We also fixed in Zenkers too. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303)

RE: [Histonet] PPE's embedding and cutting

We wear gloves to section, not necessarily for safety reasons but to eliminate squamous cells on the slides. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to

RE: [Histonet] static

Why don't you ask them to sit down at scope and show you what they are seeing? That's the best way to approach any staining or sectioning issues. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303)

RE: [Histonet] Help...

Fawn CAP has specific quidelines, its right there in the questions, did you do what that question is asking and have you documented what was done. They tell you right in the check sheet what needs to be completed. Test validation must be performed on a minimum of 25 cases (they recommend 25-

RE: [Histonet] gurney cover

I'm not sure of a plastic transparent one, but MOPEC sells the covers. This goes way back in the early 90's but I think they have a person that makes them for them, so they may possibly be able to make you one that is transparent. Its worth a try and a phone call. Liz Elizabeth A. Chlipala, BS,

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