RE: [Histonet] Tap Water or filtration system

We use tap water and have had no problems. Sydney tap water seems to be quite innocuous Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road

RE: [Histonet] RE: Saponin Technique

. Remove supernatant and add equal volume of lysis solution. 3. Resuspend and incubate for 5 minutes at 4oC. 4. Centrifuge, if blood still remains, then repeat from step 2. 5. Resuspend pellet in Hanks. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory

RE: [Histonet] PAS staining

decreases, you will notice a gradual loss of glycogen staining. Decrease in PAS staining of Fungi and basement membranes is often not as apparent. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's

RE: [Histonet] DIF Transport Media

of plasma membranes and intracytoplasmic organelles occurs after 48 hours storage. On the other hand, antigenicity is well preserved even after many days storage (Fischer 2006). Fischer (2006) Intern J Surg Pathol 14(1):108. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory

RE: [Histonet] Re: Marking inks

) J.Clin.Pathol 43:346 Hunter-Craig et al (1991) J.Clin.Pathol. 44:874-875 Patterson Davies (1988) J.Clin.Pathol 41:1013-1016 Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr

RE: [Histonet] ANP. 23075

Hi all, Weekend fridge temps are a problem. For critical fridges (Mortuary, -70oC, antibody etc), I would attache the temp logger to an alarm so if the temp goes out of range, you will be warned out of hours and have time to save the day. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys

RE: [Histonet] After hours autopsies

Richard, We do not do out of hours autopsies (too costly). We will only collect samples for metabolic studies (muscle, liver, skin) out of hours (since they need to be collected asap post-mortem (and then you probably knew that!)). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC

RE: [Histonet] HE+ Alcian Blue

is. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message

RE: [Histonet] modified Mayers Hematoxylin

5min. 4. If required, counterstain in Eosin solution. 5. Dehydrate, clear and mount. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road

RE: [Histonet] please enter

Prognostic factor J Am Soc Nephrol 13(1):234-41. 8. Nickeleit, V., Zeeiler, M., Gudat, F., Theil, G., Mihatsch, M.J., (2002) Detection of the complement degradation product C4d in renal allografts: diagnostic and therapeutic implications: J Am Soc Nephrol 13(1):242-51 Regards Tony Henwood

RE: [Histonet] RE: Histonet Digest, Vol 80, Issue 31

. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original

RE: [Histonet] PAP

carbonate. Mix well. Store solution in dark brown, tightly capped bottle. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street

RE: [Histonet] Formalin substitutes, anyone?

Hi Michael No we do not use formalin substitutes Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001

RE: [Histonet] Formalin substitutes

protect bugs from formalin. 2. Formalin dilutes as it penetrates tissue. 3. Formalin substitutes may not be germicidal. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital

RE: [Histonet] Brain Problems

The fixation time is too short. Fix as long as you can (how about 2-3 days at least for brain?) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road

RE: [Histonet] DISTILLED WATER FOR BUFFER FOR AUTOMATED STAINER

capacity of the solution. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

RE: [Histonet] Reprocess

(Johnson 2003). Johnson (2003) Histologic 36(1):21-22. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag

RE: [Histonet] DAB

fifty 1ml reagent vials 4. Aliquot 0.1ml DAB solution in each tube 5. Freeze and Store at -20oC For use: Add 0.1ml DAB solution to 50ml buffer and add 50 microlitres of 30% hydrogen peroxide. I prefer to use the DAB tetrahydrochloride Regards Tony Henwood JP, MSc, BAppSc

RE: [Histonet] Help Resurrecting Dried Fixed Tissue

. Replace one third of fluid with 95% ethanol, repeat until tissue becomes firm. Reference: The histological examination of Mummified Tissue Stain Techn 30:277-283, 1955 Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax

RE: [Histonet] tumour,lymph node and prostate preservation for histology

, paraffin embedded tissues that have been stored at room temperature for 40 years! (and others have also shown this). Contact me for further details. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318

RE: [Histonet] Forwarding Request for Control Mehtod

Ah yes, I remember, actually I don't. It was a bit of a blur!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead

RE: [Histonet] Forwarding Request for Control Mehtod

for two or more days in NBF, process as usual. Voila - fungi controls Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead

RE: [Histonet] Re: How to troubleshoot a cytology slide withdehydration problems

-hydration step. I have found that slides that have been air-dried for longer than two days do not rehydrate well. Cells appear pink with large air-dried, pale staining nuclei - lacking chromatin details. Unfortunately there is little you can do in these cases. Regards Tony Henwood JP, MSc, BAppSc

RE: [Histonet] IHC Slides, Hotplate vs Oven

that for a sweeping statement! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145

RE: [Histonet] IHC Slides, Hotplate vs Oven

We use an oven (62oC) for 25min only. See: Henwood, A., (2005) Effect of Slide Drying at 80°C on Immunohistochemistry J Histotechnol 28(1):45-46. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318

RE: [Histonet] frozen lung sections

this previously on some tissue that was received the same way. It seems that the OCT had freeze-dried prior to receipt in my lab. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital

RE: [Histonet] Removal of carbon pigment

Muhammad, Unfortunately you are stuck with the carbon. No technique (short of removing the cells/tissue section) will remove the carbon. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's

RE: [Histonet] questions

Dorothy, Apart from hoping the blocks of tissue are not too thick, we microwave the cassettes in 10%NBF in a Milestone Mega TT - 2 hours at 45oC. This ensures adequate fixation prior to processing on a Shandon Excelsior Tissue Processor. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys

RE: [Histonet] Removing tissues from OCT for molecular analysis

Katie, Rinse the frozen OCT surrounded tissue in Hanks or similar cell culture fluid. Two to three times should be enough to remove the OCT Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's

RE: [Histonet] freezing mouse heart tissue

on brain intraoperative (frozen section) Histology J Histotechnol 30(3):193. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street

RE: [Histonet] Removing Yellow color from slides refixed in Bouin'ssolution

Place slides in saturated lithium carbonate for a few minutes. This should do the trick Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road

RE: [Histonet] Masson's Trichrome, Weigert Hematoxylin Equivalent

, Nader A, Rittman B R (2007) Celestine blue and hemalum for staining nuclei Biotechnic Histochemistry 82(3): 167-168. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr

RE: [Histonet] light staining

pale after the eosin, then the acid in the eosin is continuing the differentiation process. Extend your Hx staining time decrease the eosin time. If it is a young Hx then you might find that the staining improves with time (ie it ripens). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT

RE: [Histonet] Tips for drying slides before HE stain.

I would suggest 65oC for 20-25 minutes. High temperature drying will affect IPX results and, especially with under-fixed tissues, adversely affect HE staining. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845

RE: [Histonet] Re: Diff-Quik

of the most dedicated professionals around and believe that our profession is heading in the right direction. Robert's comments should be heeded and pondered. I definitely did. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612

RE: [Histonet] Re: Diff-Quik

Robert, I agree with your comments. There are some labs that never look at their slides (the factories) but there are others (and this number in my experience is increasing) where the technologists often solve the problems before the slides leave the lab. Regards Tony Henwood JP, MSc, BAppSc

RE: [Histonet] pH of 10% NBF

is debatable. Carson et al (Am J.Clin Pathol 59:365, 1973) recommend that Millonig's Phosphate Buffered Formalin should have a pH of 7.4 and the effective osmolarity is about 290mOsM. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306

RE: [Histonet] FNA stain

to give an immediate provisional diagnosis at the time Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag

RE: [Histonet] masson trichrome - question

No Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original

RE: [Histonet] !0% buffered formalin corrosive?

I would consider formalin corrosive regardless of the form it is in. It is cytotoxic. That is why it kills and preserves cells and tissues so well. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318

RE: [Histonet] Fwd: foxp3 ab ihc

Marilyn, We have used Biolegend's Rabbit anti FoxP3 (Cat No 623801) at a 1/20 dilution with citrate retrieval (ER1) on the Bond Max with very good results. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318

RE: [Histonet] Schiff mystery

the positive blue staining. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

RE: [Histonet] Orienting GI biopsies during embedding

I would recommend you try to embed them on edge. Even if very small, you will be surprised how many are correctly embedded Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital

RE: [Histonet] Steiner and Steiner

nitrate in the Steiner techniques and found the resulting spirochete staining to be as good as or better than when uranyl nitrate was used. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's

RE: [Histonet] cap survays

, clinical demands etc). When a department runs this way, it does not take long for the pathologist to realise the invaluable resource their labs are. In fact they become quite proud of it! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612

[Histonet] See you at the NSH

Hi all, Just a note to say that I will be leaving next week for Birmingham Alabama for the NSH conference. It has taken me 30 years to get to a NSH and I am really looking forward to it. I hope to see you all (I will be the old, short, bald guy with the Aussie accent) Regards Tony Henwood JP

RE: [Histonet] Separation Artifact

Dear theci...@yahoo.com (whoever this is) I would suggest that to fix the problem, make sure you fix the tissues. Shrinkage is often due to alcohol fixation rather than formal fixation Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612

RE: [Histonet] Re: Permount question

of the coverslip. I would review a sample of slides one and two years old and record the quality (another audit!) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr

RE: [Histonet] antibody validation

diagnosis) Record the results in your lab notebook and YOU ARE Done but, Continue to monitor the antibody's performance - this is important. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's

RE: [Histonet] FNA SLIDES

, then second pass for cell block. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145

RE: [Histonet] RE: Trouble making a good Cell Block

Maria, I did not know whether you had many responses. It seems that our Histonetters are forgetting to Reply to all so that their valued knowledge is made available to all of us. Please everyone, reply to all not just the sender Thanks Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys

RE: [Histonet] in situ with paraffin sections

by researchers because they seemed like the right thing to do not because they were required. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road

RE: [Histonet] Amyloid staining on frozen muscle bx's

one of the metachromatic dyes (eg Crystal Violet)? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001

RE: [Histonet] destaining cyto slides for immuno

antibody and proceed as usual. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145

RE: [Histonet] Tissue Washing

the paraffin Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

RE: [Histonet] Please Help!

Nick, NBF may not work for Delia. She needs to do IF on the frozen sections and NBF fixation will require her to do AR on the sections (esp if she is localising immunoglobulins and complement). Possibly warming the frozen tissue, using a thumb, may help. Regards Tony Henwood JP, MSc, BAppSc

RE: [Histonet] tissue submission question

or on the cold shelf in the cryostat. It is important throughout this process to not let the tissue thaw too much. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury

RE: [Histonet] Artifact in Muscle bx's frozen for extended time

, Elisabeth J. Rushing Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

RE: [Histonet] Hi Jan,

accent. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original

RE: [Histonet] VonKossa's calcium stain

stained calcium deposits should then stand out quite well. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag

RE: [Histonet] form for internal and external errors and correctiveactions.

action taken: Review of corrective action: Issue raised at management meeting: Y/N Further action required?: Y/N CAR completed?: Y/N Signed: .. Date: Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys

RE: [Histonet] Preservation/transport medium for tissue culture

Diana, I would use Hank's solution. Sigma Cat No.H9269-100ml Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead

RE: [Histonet] Leprosy controls

. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message

RE: [Histonet] Storage of tissues in PFA before dehydration andembedding

with under-fixed tissue: Gomes L, Mackie N, Catchpoole D, Henwood T (2008) Test Your Knowledge J Histotechnol 31(4):138-184 Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr

RE: [Histonet] Storage of tissues in PFA before dehydration andembedding

fixation step altogether. AND how do you fix it in 4% polyformaldehyde? Surely when it is in solution it becomes formaldehyde? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead

RE: [Histonet] Peroxidase block again................

Morphol 2008;16:393-399 - suggesting that microwave exposure (HIER) is able to block the endogenous peroxidase. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr

RE: [Histonet] muscle striations

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message

RE: [Histonet] Peroxidase block again................

Yep, You can use alkaline phosphatase labelling instead of peroxidase Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street

[Histonet] RE: [IHCRG] CD21

: 605-611 Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA

RE: [Histonet] IHC control debate

Here is one reference that might be of use: Philippe Bertheau Variability of immunohistochemical reactivity on stored paraffin slides J Clin Pathol 1998;51:370-374 Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612

[Histonet] RE: [IHCRG] HRP-labeled primary antibodies

method for formalin fixed, paraffin embedded renal biopsies: a comparison with the peroxidase labelled streptavidin/biotin method J. Histotechnol 19(2): 125-129. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845

[Histonet] RE: [IHCRG] Re: CMV antibody

Interestingly, This data sheet does not seem to list the clone of the CMV monoclonal antibody, or am I misreading it? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead

RE: [Histonet] cloudy formalin in tissue processor

Possible carry over of xylene from the flush program? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag

[Histonet] RE: [IHCRG] Re: CMV antibody

Michael, I have used both the Dako (Clones CCH2 + DDG9) and the Novocastra products (Clones 2 and 6) and they both stain the same. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital

[Histonet] RE: [IHCRG] CMV

Jean, No the virus is the same for IPX purposes. I have used both the Dako (Clones CCH2 + DDG9) and the Novocastra products (Clones 2 and 6) and they both stain the same Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306

RE: [Histonet] NADPH-diaphorase

Eab, We purchase our NADH (reduced form) Cat No. N8129 from Sigma-aldrich Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street

RE: [Histonet] Masson Trichrome

a little pink still remains in the collagen since the acidic blue or green solution will continue to differentiate the red muscle stain. The blue collagen solution stains faster than the green solutions so use for a shorter time. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC

RE: [Histonet] container for undiluted bleach

Why do need to store or use undiluted bleach? I would dilute it to a working solution and use that. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road

RE: [Histonet] TBS vs PBS

I am not sure of the effect of buffer type on the protease but it might be possible that the proteinase K is the problem. My experience is that they can vary between lots. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax

RE: [Histonet] 5 Reasons

Cindy, A much respected and knowledgeable Histotech (who is now retired) said: Histotechnology - you either hate it or love it with a passion! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318

RE: [Histonet] FREEZY spray

Hey, Would you believe that putting the word Taupin into my outlook rules actually works. Now every obnoxious post from Taupin actually disappears into my Junk folder and I do not have to read the drivel. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager

RE: [Histonet] Colloidal Iron control

staining a suggested chromophobe renal cell carcinoma, a block containing normal renal cortex, along with the tumour, be selected. Slides are acceptable for interpretation only if the glomerular mesangium stained distinctly and the tubular epithelium showed no staining. Regards Tony Henwood JP, MSc

RE: [Histonet] Unsubscribe

unfortunately have to regularly use it) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145

RE: [Histonet] gi microwave processing

temperatures. GI biopsies being quite small do not need over-the-top processing, but in my experience, fixation is still the key. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead

RE: [Histonet] AE1/AE3 nuclear staining

Possibly the concentration of hydrogen peroxide in the DAB solution has been slowly increasing (maybe micropipette creep?) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital

RE: [Histonet] I'm outta here

But just use the delete key. I do Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW

RE: [Histonet] Burned Tissue

I would suggest checking the fixation. We had similar problems if the tissue was not completely fixed. Are the tissues free to move around in the cassettes (ie have free unhindered access to the fixative)? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager

RE: [Histonet] FW: formalin neutralizers

formalin turns purplish with the addition of Schiff's reagent, it is not totally neutralized and you will need to add more ammonia. 10. Dispose of appropriately Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612

RE: [Histonet] Quality Improvement

. The best method of documenting quality control. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001

RE: [Histonet] Water...I need more input

days to deionize water. To produce pure water, boil 600 ml of deionized water and add 6 mg of potassium permanganate to it and distil the water. Discard the first 100 ml of distillate. The next 300 ml will be very pure water. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC

RE: [Histonet] Paraffin Block triming

We also have a paratrimmer and agree with Pam Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001

RE: [Histonet] Paraffin Block trimming

No, We find them faster and we believe safer Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001

RE: [Histonet] Does anyone know why skin sections do not seem to take up haematoxylin as well as other tissues. The chromatin detail is not as good as with other tissues.

. This will affect the haematoxylin staining. To improve the staining, try pretreating the dewaxed sections with 1% periodic acid for 10 minutes prior to HE staining. This might work. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306

RE: [Histonet] RE: DAB

peroxide § To specifically demonstrate eosinophil peroxidase add sodium azide to the incubation medium to a concentration of 0.005% Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's

RE: [Histonet] Storage of muscles

Minus 70 degrees A normal minus 20 freezer tends to dessicate the tissue Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street

RE: [Histonet] sodium 5.5-diehylbarbiturate

. Loughlin, M. (1993). Muscle biopsy. A laboratory investigation. Butterworth-Heinemann pp.78-79. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road

RE: [Histonet] query regarding IHC slide

Look at the mountant you are using. Some are not as colour-fast as one would expect. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth

RE: [Histonet] bluing in tap water?

(indicating that carry-over haematoxylin is in its acidic state) then the pH will be acidic and need replacing. How can you tell if the Haematoxylin is blued? Check microscopically. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306

RE: [Histonet] Question to all my colleagues!

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -Original Message

RE: [Histonet] Re: Masson trichrome stain

We have had great success with the simple Van Gieson stain. It stains the collagen ( fibrosis) really well. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr

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