Re: [Histonet] Alcianblue for fast frozen sections

Hello Gudrun, Alcian blue for fast frozen sections I was not aware that alcian blue would be routinely used on frozen sections of transplant liver biopsies. There are other staining techniques that contain a blue dye, that have been used in liver biopsies including Masson's Trichrome (aniline

Re: [Histonet] IHC background

Paula, What does the artefact look like? Another cause could be partial lifting of parts of the section, allowing reagents to become trapped under the section. You commonly see it in thyroids where the colloid traps antibody and/or detection reagents under the colloid giving a particulate

Re: [Histonet] Clarification about Immunohistochemistry

"Bond wash" is the propriety buffer used in the Bond Immunostainer. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612

Re: [Histonet] Clarification about Immunohistochemistry

Hi there, I have had good results with Giemsa-like counterstaining (Stefanović et al 2013, Ravishankar et al 2016) : Azure blue, when substituted for hematoxylin as a counterstain in immunostain preparation, has been used to help differentiate melanocytes from melanophages. Azure blue

Re: [Histonet] Histonet Digest, Vol 220, Issue 8

Hello Jayalakshmy, iPassport is our digital quality management system. It is a commercial product that allows us to collect data, record audits, document Corrective Actions, manage controlled documents (eg manuals), expiry data and verification of antibodies, staff competencies and training

Re: [Histonet] Histonet Digest, Vol 220, Issue 8

Hi Jayalakshmy, This our policy at the Kid's hospital in Sydney: Validation of Expired Antibodies Usually when a new concentrated antibody is received it will have an expiry date of around 2 years from receipt but usually we can continue to use antibodies well past this expiry date. If the

Re: [Histonet] Dr Freida Carson

I agree, She will be missed Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the

Re: [Histonet] tissue disposal

Hi Pam, We have used the Milestone Tissue Safe Plus unit with Integrated Data Logger card reader to seal formalin-fixed placentas (and other specimens if needed) since 2019. Placentas are drained of excess formalin in a chemical hood, placed into a labelled bag and sealed. So possibly for

Re: [Histonet] Vacuum packing of formalin fixed tissues for archiving

Hi Michelle, We have used the Milestone Tissue Safe Plus unit with Integrated Data Logger card reader to seal formalin-fixed placentas since 2019. Placentas are drained of excess formalin in a chemical hood, placed into a labelled bag and sealed. We instituted the process to reduce the

[Histonet] Fw: Fixative in diff-quick

Hi Clay, The first solution is methanol. So lab grade methanol will do. The blue colour is from an innocuous dye added to differentiate from water (and ethanol used in cytology fixation). This helps us poor cytologists when we are doing ROSE at FNAs. Regards Tony Henwood JP, MSc, BAppSc,

Re: [Histonet] Jones' Methenamine Silver Stain for Basement Membranes of Kidney - Issues and Questions

I agree with Bryan, The introduction of thiosemicarbazide before the silver step improves the staining immensely. I would also look at the periodic acid. Is it too dilute, though 0.5% should work? I usually cover this by using a 1% solution for 20 minutes. Regards Tony Henwood JP, MSc,

Re: [Histonet] Retirement in sight!

I am reminded of the Beatles classic: "Will you still need me, will you still feed me, When I'm sixty-four" 40 years! - a sh.. load of experience, a sh.. load of knowledge. Histotechnology worldwide still needs your wisdom so we hope you can keep an eye on us through Histonet and the Block.

Re: [Histonet] New cryostat validation procedure

Hi Greg, I would just grab some kidneys from the butcher, freeze and section as usual. They should give equivalent results to the older unit. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct Fellow,

Re: [Histonet] cytology non-gyn screening

do your cytotechs screen the non-gyn cases? Yes Do they regularly assist with FNA's? Yes How many paps are they expected to screen in a day? Not as important as the Turn-around-time (TAT) - if the TAT is short, regularly, then all is good. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys,

Re: [Histonet] frozen section problem

We have had this issue previously. We tracked it down to the brain biopsies arriving in "Isotonic" saline (which is really not isotonic). See: Henwood, A., (2007) “Adverse effect of saline on brain intraoperative (frozen section) Histology” J Histotechnol 30(3):193. Ask the surgeons to send

Re: [Histonet] "cooked" biopsy

Hi John, There is anecdotal evidence that recycled xylene is of a better quality than the original purchased product. We have found it to be so. No processing nor staining problems unless there has been a recycling issue (as shown by the CBG xylene purity test). I suppose we have to access

Re: [Histonet] "cooked" biopsy

Hi Paula, We can check the purity of the xylene quite easily: Xylene Purity Test Procedure Note: The recommended and most accurate method of determining the purity of the recycled xylene is by doing a Gas Chromatography analysis. The following method can be used to obtain an acceptable

Re: [Histonet] Unstained slides precut for IHC

Hi Carrie, Over-heating of sections (and tissue blocks) for IPX is probably one of the most significant and unappreciated pre-analytical factors that can affect immunolocalisation. We closely scrutinise slides closely for overheating, especially those sent to us for immunostaining. We strongly

Re: [Histonet] Prussian Blue Reaction

The following might be useful: Iron Histochemistry - A Review It is convenient to divide iron-containing complexes in human tissues into two categories: those in which the iron is loosely bound to proteins and easily released by mild acid treatment (eg hemosiderin) and those in which the iron

Re: [Histonet] Teaching color blind Histo student

With Red-Green colour-blindness, I have found that a multi-stain approach works for example: Often the ZN counterstain used is light green. A second ZN stained with Loefler's Methlene Blue for comparison seems to help. The same can be done with the Masson's Stain (compare the Fast Green FCF or

Re: [Histonet] IHC with zamboni's fixed tissues

Hi Colleen, I was involved in an Immunohistochemical study of the nerves in the human ureter that used Zamboni's fixative and cryostat sections. This study was before the invention of HIER. If you are using Zamboni-fixed, paraffin embedded tissue then you will probably need to do HIER before

[Histonet] Source of Bombesin antibody for IHC

Hi all, Our usual supplier of Bombesin antibody (ABCAM AB86037) (used for neuroendocrine cell hyperplasia of infancy) no longer carries this antibody. Does anyone know of an alternative that works well in FFPE sections? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC),

Re: [Histonet] Dipp Kwik Stain

There is a generally accepted scheme of staining which is expected of Romanowsky stained preparations, namely purple chromatin, blue leucocyte cytoplasm, purple-black basophil granules, red-pink eosinophil granules, purple neutrophil granules, purple platelet granules, and pink red-cells (1).

Re: [Histonet] hexamethyleneimine

I agree Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the children's hospital

Re: [Histonet] JB4 embedded specimens

I have found that that this gives a pleasing tinctorial result: Haematoxylin and Eosin Principle Traditional HE procedures generally worked poorly on GM sections, with background staining of the plastic embedment and the loosening of sections from glass slides when alcoholic eosin is used.

Re: [Histonet] Fwd[2]: Re: AFIP Manual

Hi Maxim, Excellent At least this amazing piece of history can live on. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax:

Re: [Histonet] Formalin fixation for COVID-19 positive tissues .....

Hi Richard, It will depend on the size of the tissue and the source. Lung tissue is the major concern. Other tissues not affected as much (based on the burgeoning literature on Covid-19). Routine fixation time are applicable, remembering that the alcohols and heated wax will also inactivate the

Re: [Histonet] Recommended thickness of Amyloid sections

I agree, Slightly thicker makes the polarisation easier to see (personal experience). I would love to see a study comparing section thickness Vs polarisation characteristics. Anyone interested? (being a kids hospital we rarely see amyloidosis) Regards Tony Henwood JP, MSc, BAppSc,

Re: [Histonet] On-line references

And each edition gets better (and better). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology

[Histonet] Flippin on the Bond

Hi all, I was reading an article entitled “Evaluation of Tissue Adhesion and Staining Performance of BOND Adhesive Microscope Slides: A Comparative Study on the BOND-III and BOND-MAX Platforms” (https://pdfs.semanticscholar.org/f5f1/ad0741bc5445d0e1079a0d07a32c8a3a26c1.pdf) and they make

Re: [Histonet] Frozen section slides fading

Hi Dane, Fading and especially leaching of the eosin, will occur if slides are incompletely dehydrated. Flip out the subsidiary condenser lens (or drop the condenser) and look for refractile water droplets. Certain mountants can also cause fading of H I would also be concerned about the xylene

[Histonet] Fw: [EXTERNAL] Pap slides

We regularly leave smeared slides in 95% overnight and often over the weekend before staining. There have not been any morphological issues noted. Remember to seal the lid since alcohol will evaporate quite quickly. You will notice an increased eosinophilia in the dried upper area of the

Re: [Histonet] Need a procedure

he patients sample.Think about that. > > Didn’t know about the heat issue with Gel. Important to know. Thanks ! > > Garrey > > Sent from my iPhone > > > > On Jan 23, 2020, at 6:12 PM, Tony Henwood (SCHN) via Histonet > > histonet@lists.utsouthwestern

Re: [Histonet] Need a procedure

Hi all, Be careful of using cell block matrix that requires heat to solubilise the matrix (eg agar or other commercial matrixes like Histogel). Adding a heated matrix to unfixed, or even formalin fixed, material can denature some antigens (eg CEA) resulting in a false negative IPX.

Re: [Histonet] Cell block preparations

Hi Jennifer, I have had excellent success with lysing the red blood cells (using Isotonic Ammonium Chloride) prior to cell block preparation with thromboplastin-plasma. The lysing solution contains EDTA so you will need to add a few drops of 1% calcium chloride. Method as follows: Lysis

Re: [Histonet] H protocols

Hi Charles, Yes we use the same protocol for all, though sometimes we need to modify if for example tissue has been overly decalcified (eg mandible). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at Westmead Adjunct

Re: [Histonet] Delay in embedding?

Hi Paula, We routinely do this, especially for our fetal autopsy blocks. We are then able to process and let them set at room temp until we are able to embed and cut. Some cases are more urgent than others so these can be expedited a lot easier since they will only need embedding, sectioning

Re: [Histonet] Cell block processing

Hi Charles, I have had excellent success with lysing the red blood cells (using Isotonic Ammonium Chloride) prior to cell block preparation with thromboplastin-plasma. The lysing solution contains EDTA so you will need to add a few drops of 1% calcium chloride. Method as follows: Lysis

Re: [Histonet] waterbath/paraffin

Are the tissues taken directly from the molten wax and placed in the wax-containing mold or are they allowed to cool before embedding? One cause of tissue separation is the difference in temperature between tissue and embedding wax. Try taking directly from molten wax and embedding directly in

Re: [Histonet] Metals

_________________ From: Tony Henwood (SCHN) via Histonet mailto:histonet@lists.utsouthwestern.edu>> Sent: 30 September 2019 07:22 To: Sanders, Jeanine (CDC/DDID/NCEZID/DHCPP) mailto:j...@cdc.gov>> Cc: Histonet mailto:histonet@lists.utsouthwestern.edu>> Subject: Re: [His

Re: [Histonet] Metals

Two good screening stains are Mallory and Parker’s Fresh Hematoxylin Stain for Metals and Timm’s Silver Sulphide Method for Metals. Malloy's results: Aluminium Blue-black Copper Greenish-blue IronBlue-black LeadBlue ZincBlue For more specific staining: Aluminon Stain for

Re: [Histonet] Victoria blue for lung tissue

Whoops, The Ventana does not use Victoria blue, my mistake. They have a resorcin fuchsin solution (known as Hart's method) not Miller Victoria blue. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct

Re: [Histonet] Victoria blue for lung tissue

Methods found here: https://www.ihcworld.com/_protocols/special_stains/miller's_elastic_ellis.htm http://www.histosearch.com/histonet/Dec98/Millersstainforelasticfib.html https://www.sakuraus.com/getattachment/774d89be-4b32-4d69-bb02-8dea75c52c0c/858 ? Regards Tony Henwood JP, MSc, BAppSc,

Re: [Histonet] Victoria blue for lung tissue

Victoria Blue is the dye used in Miller's Stain for Elastic Tissue. It is also used in the Roche Ventana Benchmark Stainer to stain elastic Tissue: Miller PJ (1971) An elastin stain. Med Lab Technol 28, 148–149 Karen Percival & Zaher Radi (2017) Comparison of five elastin histochemical stains

Re: [Histonet] Processing Schedule- ASP-6025

Processing seems adequate. After processing, how long do they sit in the embedding centre block holding tank before embedding? We found that quite a few antigens were affected when we stored control tonsil in the embedding centre (dry) at 60oC for a few days before embedding. In summary:

Re: [Histonet] Derm IHC question

Possibly, the edges have been allowed to dry prior to immersion in fixative. Also is there evidence of cautery artefact? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine,

[Histonet] FW: ER/PR question

This paper might be useful: Ehinger, A., Bendahl, P. O., Rydén, L., Fernö, M., & Alkner, S. (2018). Stability of oestrogen and progesterone receptor antigenicity in formalin‐fixed paraffin‐embedded breast cancer tissue over time. Apmis, 126(9), 746-754. Regards Tony Henwood JP, MSc, BAppSc,

Re: [Histonet] Opinion on dye on biopsies

One article that might be useful is: Surprenant D, Garib G, Hutchens K, Dreifke M, Speiser J, Winterfield L, Peterson A, Krol C, Adams W, Tung R. Novel Use of Preoperative Epidermal Coloring of Very Small Dermatological Specimens-Protocol for Reduction of Lost Specimens. The American Journal

Re: [Histonet] IHC-H HELP...

Yep, Definitely an issue. You can easily stain the IPX slides with H, though the discernibility of the cells and tissue structure with the H will depend on the degree of DAB product laid down (eg I would expect it to be difficult with a Vimentin IPX compared to a CD15). (Grosset, A. A.,

Re: [Histonet] IHC Negative reagent controls

Hi Cayman, Unfortunately, applying HIER to a negative control for an antibody that requires enzyme retrieval (or no retrieval at all) is not appropriate. The pre-treatment processes are different and could unmask different epitopes. If you are using a negative control then the whole procedure

Re: [Histonet] Alizarin Red Stain

Hi Richard, We sure do Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the

Re: [Histonet] H for H. pylori?

The following might be useful: Tazawa, K., & Tsutsumi, Y. (1998). Effect of prolonged staining with hematoxylin on detecting Helicobacter pyiori in hematoxylin‐eosin‐stained gastric mucosa. Pathology international, 48(6), 448-452. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys,

Re: [Histonet] AFB STAIN

Cross-contamination has not been proven in FFPE tissues. I have not seen it in nearly 40 years of practice Do you have any evidence of cross-contamination in FFPE? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children’s Hospital at

Re: [Histonet] AFB STAIN

We do, It works well Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department the

Re: [Histonet] Unstained slides - how long are they good for?

Will definitely depend on the antibody you are using. Some references: Jacobs, T. W., Prioleau, J. E., Stillman, I. E., & Schnitt, S. J. (1996). Loss of tumor marker-immunostaining intensity on stored paraffin slides of breast cancer. JNCI: Journal of the National Cancer Institute, 88(15),

Re: [Histonet] Iron Stain

Try your eosin stain used in your H or 1% Neutral red in 2% acetic acid Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax:

Re: [Histonet] pH of formalin

We routinely use the following Hew-Shue (1991) has described a useful pH indicator for working neutral buffered formalin solutions. Bromocresol purple (CAS: 115-40-2) , when added to formalin solutions, serves as an indicator of pH as well as, from a safety aspect, labelling the solution as

Re: [Histonet] Acid Phosphatase Protocol

Hi April, Try this Control tissues: liver, kidney and prostate Fixation and Sectioning: Air dried unfixed 8µm frozen sections Solutions: 1. Substrate solution Naphtol AS-B1 phosphate 0.005g (Fluka 70494 1g) Dimethylfomamide0.5ml Warning: Suspected

Re: [Histonet] Elastic stain

I would recommend the Orcein stain: Henwood, A., (2003) "Current applications of orcein in histochemistry. A brief review with some new observations concerning influence of dye batch variation and aging of dye solutions on staining" Biotech Histochem. 78(6):303-8. And you can pair the orcein

Re: [Histonet] validation

Immunohistochemistry validation is much more than simply 20 positives and 20 negative cases, though this will allow you to meet most accreditation requirements. Validation is a multi step process: 1. Optimisation: following a thorough literature review (what should and should not stain,

Re: [Histonet] Adenovirus antibody?

Hi Paula, We use Adenovirus antibody from BioSB (Diagnostic Technology) Cat Number BSB 5040 Clone 20/11 and 2/6 Isotype IgG1/K Dilution1:200 with citrate retrieval on the Bond 3 Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the

Re: [Histonet] IHC questions

Hi Vicki, We recently did a study that we published in our local journal (Histograph June 2017) that migh be of use: Controlled Section Baking for Immunohistochemistry Tony Henwood, Principal Scientist, Histopathology, The Children's Hospital at Westmead One source of poor immunostaining

Re: [Histonet] PAX-5

Hi Beth, We have found the same thing. You get a moderate staining (half of what you see with most B cell lymphomas) with clone 1EW (as supplied with the Leica Bond rtu). You will get similar results with clone 24 (38%: Adams, et al (2009). Clinical, phenotypic and genetic similarities and

[Histonet] Method for Patzelt stain

Hi all, I am having difficulty locating the method for the Patzelt stain. I came across it while reading: Kimura, S., Hirai, A., & Shimizu, H. (1981). Epidermal vacuolation: an artifact due to injection of local anesthetics. Archives of dermatological research, 270(4), 413-419. This paper

Re: [Histonet] Inhibin

We use Leica rtu PA0110 on the Bond 3 Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology

Re: [Histonet] IHC testing for Parvovirus (Human)

Hi Richard, We use NCL-PARVO from Leica Details as follows: ANTIGEN NAME:Parvovirus B19 OTHER NAMES: Clone: R92F6 Isotype:IgG1 DESCRIPTION: Mouse monoclonal to native parvovirus B19 purified from human plasma. Human parvovirus B19

Re: [Histonet] A Question About Paraffin Times

It is best to remove the blocks and let them set at room temperature. Leaving them at 60 or more degrees can adversely affect antigens. We routinely do this with our autopsies and have not noted any issues. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal

Re: [Histonet] Cytology pap staining

I would also check the fixation. Do the smears look air-dried. The larger nuclei, following air-drying do not concentrate the Hx as much as prompt alcohol fixation resulting in paler stained nuclei. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal

Re: [Histonet] processor died overnight

Yep start at last alcohol in other processor, xylene & wax as usual. If the tissues have remained covered in alcohol, you should not have a problem Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct

Re: [Histonet] help!!

: Re: [Histonet] help!! Tim and Tony, Why couldn't DAB be used on frozen sections in your example? Allan On Mon, Apr 17, 2017 at 9:03 PM, Tony Henwood (SCHN) via Histonet <histonet@lists.utsouthwestern.edu<mailto:histonet@lists.utsouthwestern.edu>> wrote: Hi Bianca, Well for most Patho

Re: [Histonet] help!!

Hi Bianca, Well for most Pathology departments, Immunofluorescence (IF) is used for Renal and Skin biopsies; looking for human Immunoglobulin (Igs) deposition on basement membranes. The advantage here is not so much the fluorescence, but that we use unfixed frozen sections. The buffer rinse

Re: [Histonet] Tissue Fixation

If the specimen is placed on absorbent paper prior to fixation, the paper will gradually absorb the water from the specimen causing it to dry out. Microscopically the tissue will look a little like what you see when a laser scalpel is used to excise a skin specimen. The heat affected eosin

Re: [Histonet] Reference for this Recipe and Use

This is sometimes known as Newel's solution for revealing lymph nodes in fatty specimens: Newell KJ, Sawka BW, Rudrick BF, Driman DK. (2001) "GEWF solution" Arch Pathol Lab Med 125:642 645. Absolute ethanol1000ml Water 340ml

Re: [Histonet] Bob Richmond

All the Best Bob, Have a great retirement. Maybe we will see you in Australia on one of those big boats that make their way here regularly. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow,

Re: [Histonet] Dirty H after break

Hi Allan, With the quality of the eosin dyes today, filtering should not be required. The same for xylene and ethanols. You will need to monitor the usage and solution levels (evaporation being the issue here). There are no hard and fast rules but a suggestion is one slide per ml of

Re: [Histonet] Dirty H after break

Hi Allan, The flakes are alum precipitates. I would suggest filtering the haematoxylin before use. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western

Re: [Histonet] Staining issues

Hi Charles, Many issues could be in play. Since the centre of the tissue is causing problems, then I would look at fixation; are the spaces around cells in the centre of the block wider than at the edges (conversely do the cells look like they are smaller than their counterparts at the edge)?

Re: [Histonet] Weak Hematoxylin Staining?

Vivian, After cutting the frozen sections, how are you fixing them? If they are air-dried then they will tend to give a washed out nuclear stain. I would suggest immediate fixation of the sections in either methanol or to make even a brighter H: 1% acetic acid in ethanol (1ml glacial acetic

Re: [Histonet] RTU antibodies with 1+ year expiration date

Hi Alan, Not every source states that diluted antibodies only last for a few weeks, in fact my experience as well as those of the commercial suppliers (see their data sheets) indicates otherwise, for example, I just re-validated an antibody (CD45RO) that was prepared 8 years ago (but

Re: [Histonet] Competency for Supervisors

Hi Mighnon, I find the so-called competency for all staff, including laboratory managers one of the stupidest items ever to be put in a standard (ie ISO 15189). All staff who perform a test need to be recorded as being competent in it, no problem with this but managers as well? How

Re: [Histonet] Transport training for formalin fixed specimens

Hi Gareth, You will probably have to design your own Continuing Education Session. I would recommend the following topics: 1. Nature of formaldehyde - safety aspects, explanation of the MSDS. 2. What formaldehyde is used for 3. Appropriate packaging of pots containing specimens in

Re: [Histonet] Melanin Bleach

The KMnO4/oxalic acid sequence will weaken the tissue adherence. If you do not have to use Peroxidase/DAB, then I would recommend alkaline phosphatase labelling (+red chromagen). Remember KMnO4/oxalic acid can be deleterious to some antigen-antibody reactions. We found that LCA, CAM5.2, L26,

Re: [Histonet] Cytology/Histology Staining Question

Yep, We do. We have a separate dehydration sequence for PAP and other special stains (separate from the eosin dehydration sequence). We have a Leica (used to be Vision Biosystems) Autostainer Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the

Re: [Histonet] PAS Stain

Only a crazy Aussie would do this!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Principal Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine, University of Western Sydney Tel: 612 9845 3306 Fax: 612 9845 3318 Pathology Department

[Histonet] PAS Stain

Yep, I have had a few techs in my time who could not digest glycogen if their life depended on it. Their salivary amylase just did not work. It was not a major (not even a minor) health issue for them. They looked healthy enough (actually healthier than me). This is one of the reasons we

[Histonet] Reference for erythrosin-eosin counterstain for H

Hi all, I am doing an H workshop next month and one of the Hx counterstains I will be including is Erythrosin-eosin (see below). This counterstain is the preferred by some of our major Histopathology departments in Australia. The issue I have is that I cannot find a reference for this variant

Re: [Histonet] Porcessing FFPE tissue without alcohol??

You might find these articles to be useful Tracy, R. E., & Walia, P. (2002). A method to fix lipids for staining fat embolism in paraffin sections. Histopathology, 41(1), 75-79. Turello, R., Snyder, D., & Hartman, H. A. (1984). A modification the osmium tetroxide post-fixation technique for

Re: [Histonet] H staining

I would look at the dewaxing stage. Have you changed waxes? Try longer in xylene &/or longer in the oven. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager & Senior Scientist, the Children's Hospital at Westmead Adjunct Fellow, School of Medicine,

Re: [Histonet] Dako Artisan special stainer

Oh dear, Are you considering sacking your Special Stains Tech? Only kidding. I wish I had the money to buy one myself. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) Laboratory Manager Senior Scientist, the Children's Hospital at Westmead Adjunct Fellow, School

Re: [Histonet] Tissue processing validation

The validation would already have been done. You only need to check if the re-location has affected any equipment components. I would take a few cassettes of spare tissue and process it on your most used program and await the results. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys,