[Histonet] Early fixatives
Ryan Hickey asks about the historical use of fixatives, and stains derived from natural sources. A good place to start is with the book by Brian Bracegirdle " A History of Microtechnique" (second edition) Published by Scientific Heritage 1987. There are many articles out there and you will have to do a literature search for them. Regards Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Thank you Dr Margraf!
Thank you Dr Margraf, Dr Cope and now Dr Sengupta for giving up your time and hosting the "Histonet". It is an ideal way for histotechnologists and others to be kept informed. And Merry Christmas to everyone from the heart of Dixie! Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Placenta's
Joyce Fortin asks about placenta's: I work in a teaching hospital with a busy L&D department and we examine every placenta from every birth. Depending on their history the placenta may be "gross in" (with four blocks) or "gross only". The physicians are happy to have a record of the weight and appearance and dimensions, and microscopic appearance Other area hospitals, however do not examine their placenta. Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Friday histology trivia
Any one who has seen the CBS evening news with Scott Pelley lately (at least in my area) will have seen an ad for a product named "Salonpas". In the background is a microscope. It is interesting that when you see a microscope in an ad on TV, it implies that real, dedicated research has been performed in bringing the product to market. In this case, the microscope looks like a 30 year old plus American Optical microscope. I don't think AO are in busincess any more. Maybe Leica took them over. Looks good though! I have never seen a microtome in an ad on TV, but plenty of microscopes! Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Telepathology
Victor Tobias asks about telepathology- I seem to remember there is a rule somewhere that primary diagnosis can only be made using a glass slide. As to chapter and verse, I don't know. However there are reports in the literarature that it has been tried out for F/S in Alaska and Finland. Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Middle School Science Day
Carol Tanck asks about material for a Middle school science day. Good items to get hold of are plastinated organs that the young people can pick up and handle. If you have a medical school or community college near by, their cellular biology or anatomy departments (Titles vary in these modern times) may be able to lend you some. These organs have been thoroughly fixed, infiltrated with plastic, and then hardened. They are safe to handle and can survive a drop or two. They are especially good for showing lung tissue from smokers showing emphysema, etc. Regards Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Melanin removal p.s.
Thats's 5 minutes in the potassium permanaganate. Some stubborn melanin may need a longer. Some authors recommend as long as 24 hours. (It's Friday - how can you tell?) Michael Titford Pathology USA Mobila AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Melanin pigment removal
Leila asks about removing melanin pigment. We run slides down to water and place in aquous 0.25% potassium Permanganate. Wash well in tap water.Rinse in distilled water. Place in aquous 5% oxalic acid for 5 minutes. Rinse in distilled water.Wash in tap water. continue with method. We run three control slides. Two of a known melanin containing slide, and an extra test slide. Treat one each of the test slides and control slides with the permanganate. The method is a little harsh. You may want to use coated slides. There are different melanins that have slightly different characteristics, requiring different times. Ref: Manual of Histological Staining Methodsof the Armed Forces Institute of Pathology (1968) Lee Luna (Edit) There are other methods to remove melanin that use potassium chlorate, and hydrogen peroxide. Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histotechnology schools in the United KIngdom
A few days ago Peggy Wenk kindly listed a web site for histotechnology schools in the United Kingdom, but it was a little mis-leading. The site was for all job vacancies in the United Kindom hospitals, all part of the British " National Heath Service". Most of the jobs listed were for physicians who wished to train in histopathology, not histotechnology. There were a few positions for what we in the USA call Medical Laboratory Technologists, that in the UK, includes histotechnology. The system in the United Kingdom is quite different to that in the United States. It may have changed recently since I left there, but In don't think so. In the UK there are no "Histotechnology schools" as such. If you wish to persue a career in histotechnology and work with human tissues, as in a hospital, you have to be State Registered. To be State Registered you have to attend their degree level national certificate program run by their Institute of Biomedical Science. It includes micro, blood banking, chemistry, hematology, etc, including histology. It is hard! During that time, you rotate through the different sections of the hospital laboratory. When you finish, and pass the test, then you can apply for State Registration. They have advanced fellowships in histotechnology. On our own Histonet from time to time, technologists from England have complained about having to work so hard and being understaffed. Maybe it is too hard for normal people!. (Histology technicians not working in a hospital or not requiring State Registration can train "on the job" and do not have to be State Registered. Years ago there was a program run by the Institute of Science Technology for non medical histology technicians that included Histology in its last two years of a five year part time program, but I don't know if it still exists) Maybe some one from the United KIngdom can bring us all up to date. Regards Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] More about the good old days..
And the stuff we poured down the sink without a second thought ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Staining Nissl
Ruth Yaskovich asks about staining Nissl: Years ago, when I started in histology and there were still dinosaurs on earth, we used cresyl fast violet for staining Nissl in celloidin sections, using a fancy differentiation with cresote in alcohol, or something. In paraffin sections you can get pretty good results staining in heated 1% toluidine blue or 0.5% thionin and differentiating in 96% alcohol. There may be some IHC antibody you can use Michael Titford USA Pathology Mobilew AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Merry Christmas, and thanks Dr Margraf!
Merry Christmas to everyone from the Heart of Dixie! And thank you Dr Margraf and Marvin Hanna for hosting the "Histonet". It is a real asset to histotechnologists. Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Mixed fiber type identification
Arjun Ruhella asked about fiber typing of animal soleus muscle: No one answered him, at least in public. I guess all the hotshots are in Vancouver! Years ago I was involved in fiber typing and measuring of human muscle biopsy fibers. In those human muscle biopsies, all the bundles were mixed, about 60:40 if I remember correctly. The proportion and size of the fibers changed depending on the disease process. We used the ATP reaction on frozen sections to visualise the type 1 and type II, (and IIA, IIB, etc) fibers. Then, in those pre-computer days, we selected a typical muscle bundle and started counting the cross sectional diameter of those type I and type II fibers, using eypepiece graticule in the microscope. A histogram was prepared. I cannot imagine using IHC to demonstrate the type I and Type II fibers is a whole lot different, or that animals have muscle fibers that are not mixed. You can see some differentiation on a good PAS stain, but I do not know how well it compares. The procedure we used was in "Muscle Biopsy: A modern approach" by Dubrowitz and Brooke, Saunders. 1973 ( I told you it was years ago!) Hope this helps Michael Titford Pathology - USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] CAP autopsy requirement?
Am I hearing right? The CAP has done away with the requirement to sign out autopsies within 60 days? (But you still have to get the PAD out in two days)? How will I ever get pathologists motivated any more?! Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cut proof gloves
Sheila asks about cut-resistant gloves: We use the cut resistant gloves from Surgipath Medical Industries. They are of course, cut resistant, but not cut proof or puncture proof. They fit nice and snugly and don't impede work. They work well if you just touch a finger lightly with a scalpel or blade during an autopsy or while grossing a specimen. When that happens the rubber glove on top gets cut and lets the blood in and you must wash the cut resistant glove (and your hand very well of course). I don't know if they are Kevlar. That may be a brand name. Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Frozen section staining question
In our frozen section room, we use plastic dropper bottles (holding about 130ml each) to pour the solutions on the slide on a rack to stain the F/S. We figure you don't risk getting floaters and the solutions are fresh. However, these bottles are "home made" and do not look professional. We have to use tape to label the bottles, and record when they were last filled, and they don't clean easily. Does any company supply a set of professionally labeled dropper bottles for frozen section stainig? I have looked in the regular catalogues (Fisher, etc) but cannot find any there. Thanks in advance Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] ATPase reaction
Hazel Horn asks about the ATPase reaction: Years ago we had problem with our ATPase reaction on F/S of muscle biopsies. We discovered that in keeping the substrate at minus 20oC in a freezer, and getting it out and letting it warm up to room temperature before weighing a small amount out for the procedure, and then putting it back in the freezer, in doing that all the time, we had inadvertantly inactivated the ATPase substrate. Its best to aliquot it out when you first receive it from Sigma, or whoever, and keep those vials frozen, getting one out to use every time you do the reaction. Other than that, I would think the solutions are suspect, too old or wrong pH or something like that. Regards Michael Titford Pathology - USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Ceramic knives for autopsies?
Has anyone had any experience with using the new ceramic knives for autopsies? Where do you purchase them? Do they hold their edge? Can they be resharpened? Thank you Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Luciferase IHC/Holy water
You can make your own holy water. You put regular water on a stove and boil the hell out of it! Mike Titford USA - Pathology Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] AFIP History
Those of you who mourn the passing of the AFIP may be interested that a book has been published about its history, entitled, "Legacy of Excellance: The Armed Forces Institute of Pathology 1862 - 2011" by Paul Stone, published by The Borden Institute, Fort Detrick, Maryland. It details the AFIP history from the Civil War period to the present. It has many, many great photographs. The early chapters are mainly about medical care in the Army during the early years, but later chapters are about its different missions in later years. There are photographs of the museum and its different specimens, teaching of pathologists, research, DNA, veterinary, forensic, etc, etc. Photographs include those of Frank Avalone, Dr Frank Johnson and Lee Luna who all devised histology staining methods. Lee Luna is described as "The Father of Histology" (He may have some competition for that title!). Dr Johnson is shown using what I think is an RCA EMU 3 transmission electron microscope, the only TEM made in the USA (I think). The National Society of Histotechnology is mentioned too. I worked at the AFIP for a few weeks on two occasions. It was interesting to see some familier faces in the book. Michael Titford USA Pathology Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Lagal cases SOP
Ann Angelo asks about releasing tissues in legal cases. I guess how you do it depends on your state and its laws, and what your own facility lawyers require. At our facility in the State of Alabama, to release tissues and/or medical records we require either a release signed by the patient, a subpoena, or a court order. If the patient has passed away, and the next of kin is making the request, we need documentsation they are legally the first next of kin. THEN, we let the lawyer in our risk management department check it over. After she has blessed it, we can release the material. A lot of requests are made by firms who go round vacuuming up every item they can for other law firms. We do not release blocks, or original slides, but can release recuts. Hope this helps. Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Formalin down the drain??
I was a little distressed to read the message from Amy in Camp Hill,
Pennsylvania declaring she dumps everything ("and I mean everything") from her
histology lab down the drain. There are a bunch of Federal Laws governing
handling and disposal of chemicals used in the histology laboratory and she
appears to be breaking several. The wastewater law limits how much formalin you
can discard down the sink (and you cannot dilute as you go). The same law
forbids disposal of organic solvents like xylene, or solutions containing
organic solvents. Local laws in Pennsylvania may be more strict.
I recommend to Amy that she purchases a book like, "Hazardous materials in the
histopathology laboratory" by Janet & Richard Dapson and read the whole thing
cover to cover!
Michael Titford
Pathology USA
Mobile AL USA
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[Histonet] And now, IHC playing cards!
Those of you that like freebies (like Dako mousepads, Thermo pens and Sigma coffee mugs) will be amused to learn that Cell Marque are giving away packs of playing cards. Each card shows a section stained with a different antobody (one of theirs of course, together with the catalogue number). No, I don't have shares in Cell Marque, but I am continually amused by the different methods companies in all walks of life use to advertise their products. Some are, well, just original! Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Ehrlich's Hematoxylin
Candice Camille asks about Ehrlich's hematoxylin- The sodium iodate to ripen the hematoxylin should be added last of all, after all the other ingredients, to ripen the stain. (So you will not see if it has dissolved, or not!). Better to ripen the Ehrlich's hematoxylin naturally. Do not add sodium iodate. Instead place the bottle in a sunny windowsill and shake it up every day. Loosen the stopper and let the hematoxylin breath. It will take a few weeks to ripen (very un-20th Century!). You can use it when it changes color to dark red to maroon color. but when ripened it will last for ever such a long time provided it is not contaminated. It gives good regressive staining and will also stain mucins. Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Failed water test
Some years ago, our deionized water system became contaminated by a gram negative organism. (I don't know what the bacteria live on, in the dark, with only deionized water to "eat"). We have a contract with a deionized water company, but they said it would be impossible to sterilize the whole system, with many labs on different floors and water lines weaving through the building and dead ends here and there. Now we screw filters on each spigot we use. The resistivity is in the right range, and the culture tests are negative. When the filter gets clogged up, the water runs slowly, and we know its time to change the filter. Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Polyvinyl alcohol
Candice Camille asks about polyvinyl alcohol. Some time ago when I was in the Army Reserve I completed a correspondence course for parasitology and it discussed PVA. I still have the manual. It states: "PVA is a mixture of fixative and water soluble resin that is specifically used to fix and preserve trophozoites of intestinal amoebic organisms. " Further on it states, " PVA is primarily used for preserving fresh specimens to be shipped to central laboratories". A formula is given. I can Fax it to you if you give me a Fax number. Regards Michael Titford USA - Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] More about "A perfect red"
The full citation is "A perfect red: Empire, espionage, and the quest for the color of desire." By Amy Butler Greenfield. Harper Collins. (It was $26.95 in 2005 but now you may be able to pick up a good used copy online). Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Colloidin? No!!
Michele Carr asks about using colloidin for cell block preparation. Years ago, histology labs would coat the inside of a glass test tube will colloidin (maybe 1% dissolved in 50:50 absolute alcohol and ether) and allow it to dry. When they needed to make a cell block, they poured the solution containing the cells in the test tube and spun it down. The aquous solution caused the colloidin to separate from the glass. you could pour off the solution and wrap the cell pellet up in the softened celloidin and put it in a cassette for routine processing. However, times have changed! Using celloidin (nitrated cellulose or gun cotton) is dangerous and the alcohol ether solvent is also dangerous. I think cytology labs use a jell called "histogel" or something in which they capture cell blocks. Maybe there is a decent cytology lab in your area that uses this product or a similier one you can try. Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Thionin stain
An Eerdekens overseas somewhere asks about thionin staining: An, you may have a bad batch of thionin stain. Did you just open a new jar? Here is the USA we use stains certified for use in histology by the Biological Stain Commission. Each lot has been tested by them for use in histology. Maybe you could borrow some thionin from another lab and see if that works better for you? Sometimes in my career I have come across bad jars of stain that just did not work. Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Acetylcholinesterase
Nicole Cosenza asks about acetylcholinesterase methods. Years and years ago in London we used Koelle's method to demonstrate cholinesterase in muscle and mouse diaphragms. The method we followed was in Pearse E, Histochemistry - Theoretical and Applied. Volume 2. Churchill Livingstone. London 1972. Pages 1312 - 1316 has several different methods. The enzyme was visualized with ammonium sulphide. The tissues were mounted in glycerine jelly. If the method worked too well and we could not see the motor end plates, we adjusted the pH to reduce staining. Dr Filipe went on to publish her own method in Filipe I., Lake B., Histochemistry in Pathology. Churchill Livingstone. London 1983 page 322. In her method, she used osmium to visualise the enzyme, instead of ammonium sulphide. Stain technology used to have articles about the method too. I have not heard of Karnovsky or Roots methods, but lot of different methods were published in the early days of enzyme histochemistry. Hope this helps Michael Titford Pathology - USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Gilson's fluid
Jorge Tornero asks about Gilson's fluid. The original Gilson's fluid was made up of Mercuric chloride 2 grams, glacial acetic acid 0.4ml, nitric acid 1.8 ml, 95% ethyl alcohol 10 ml, and distilled water 88ml. I don't know about usining zinc chloride in the formula Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Hirschsprungs disease? Try Calretinin!
Someone who did not give their name enquired about "Fetal choline esterase" for Hirschsprungs disease on formalin fixed tissue.(That word was probably "Acetyl). We used to use the old acetyl thiocholine esterase on frozen sections but switched a couple of years ago to calretinin histochemistry on routine FFPE sections. It works! We also do H&E levels as well. The reference is: Guinard-Samuel, V: Borrard, Arnaud:De Lagausie, Pascual et al. "Calretinin immunohistochemistry: a simple and efficient tool to diagnose Hirschsprungs disease" Modern Pathology 22: 1379-1384. 2009 Hope this help Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Deparaffinization for EM
Mark Elliot asks about processing FFPE tissue for EM. Hayat published a method in his book we use. Briefly you dig out 1mm cube pieces of tissue from the block and rotate them in xylene for one hour, followed by absolute, 95%, 70%,50% alcohols for 15 minutes each and then into your EM buffer overnight, followed by your normal EM processing next day. The EM image looks pretty grotty but if your pathologist can make a diagnosis it is worth it. (Hayat M.A. Principles and techniques of electron microscopy. 2ed edition. Baltimore. University Park Press. 1981 page 209) In another method published years ago, someone by-passed the hydration steps by deparaffizing in osmium dissolved in xylene, then going straight to Epon. Hope this helps Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Formalin substitutes, anyone?
I am curious!! In a Histonet message last week, Tony Henwood mentioned the formalin substitutes "UMfix", "Kryofix" (now called Neo-fix I think) and "Spuitfix". 1) How many surgical pathology labs use formalin substitutes for routine work? 2) Which ones do they use? (which is the most popular?) 3) Do they use formalin substitutes just for initial fixation, or on their tissue processors as well? I would be interested in hearing. Yes, I know, curiosity killed the cat!! Thank you to anyone who replies. Histotechnology is endlessly fascinating. Michael Titford Pathology USA Mobile AL USA = ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Technical charge 88342
Dr Cartun sent an email to the Histonet about what his institution bills for the technical component of 88342. Dr Cartun - Don't feel so guilty! In reality no one much pays top dollar for any service in the hospital. Some patients are treated as DRG patients (Hospital gets paid set amount for treating the patient) and for other patients, their insurence company has contracts with the hospital, where the insurence company (BC/BS for example) pays what they think your cost is. If your are still curious, contact your billing office and ask what Medicare pays for a 88342. Sadly, the only people who pay top dollar are often those without insurence. Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Retaining empty jars
Rhonda Hartz asks if anyone else retains empty jars after grossing. We too retain our empty jars after grossing for about a month. We keep them in a red biohazard trash bag. It pays dividends. If once the case has been signed out the physician/surgeon complains that they removed more biopsies than we listed in the report, or submitted more jars, etc, we can go back and check. On rare occasions we have found tiny pieces of tissue in the jars, or hidden in the grooves of the lid. These enquiries most often happen with tiny pieces of tissue, of course, like GI biopsies. We have the physicians write on the label on the jar what is inside it (like everyone else I guess) as well as listing it on the surgical pathology slip. You can also go back and check what they wrote down. Afterwards the jars are incinerated as they have all that HIPPA related stuff on the labels. Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Haunted cryostat/Thanks!
Thank you everyone who responded to my problem with a Leica CM 1850 cryostat turning itself off. About 11 people responded with tips. Thank you!! The Histonet is great!! In answer to some enquirys: 1) The cryostat is set to defrost at 2 a.m.. The manual says it defrosts for an hour. Jackie O' Conner asks if it is set to go back "on" after the defrost. I have no idea. It defrosts well the rest of the time, and turns itself back on. Brian Cornett-Early recommends changing the start device on the compressor. 2) Someone else asked if power is getting to the cryostat - Yes, when it turns itself off, the on/off switch is on "off". All you have to do is flip the switch and it starts pumping and cooling. 3) Mari Ann Mailhiot with Leica recommends changing a cicuit breaker on the side of the compressor. That is probably where we will start. Mari Ann works for Leica so it sounds like good advice. Thank you everyone. Since it only breaks down about once a month, it will take a long time to determine if the problem is fixed. Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Is my Leica CM 1850 cryostat haunted?
We have a Leica CM 1850 cryostat that about once every three weeks to a month, turnes itself off! Has anyone else experienced this? How do I fix it? Initially, we thought someone after hours was turning it off, maybe someone in housekeeping or a passing med tech. That was not the case. Later our Biomedical people replaced a capacitor or something in the bowels of the cryostat thinking that may be faulty and contribute to the problem. That did not help either. Still later, the cryostat was put on its own circuit (No help), and then after that, we tried an uninterruptible power source thinking minor power fluctuations were the cause (no help either). Anyone know how to fix this problem?. It is disconcerting to walk into the lab in the morning and the cryostat is at room temperature and the OR is going to have a busy day. Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] AZF
Dr Kiernan asks about AZF When the safety people in our medical center finally pried the last of the B-5 from my cold, dead, hand, we replaced it with AZF which contains formaldehyde, zinc chloride, methanol and acetic acid. Our hematopathologist claims it is better than straight zinc formalin, and, until recently, it was available in prefilled containers making it perfect for dishing out to clinics and floors etc. It works almost as well as B-5. It is sold in the USA by Newcomer supply. You still cannot pour it down the sink after use, but is not so dangerous as mercury containing B-5. Michael Titford Pathology USA Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Gomori Reticulum
Gudrun asks about how the Gomori retic procedure works: It is my understanding that the iron alum (ferric ammonium sulphate) acts as a sensitizer. The iron combines with the aldehyde formed in the reticulum from the previous oxidation step to create an aldehyde-metal complex. In the third step with the ammoniacal silver nitrate, the silver combines with this complex, but is not visible. Treatment with formlin makes it visible.Toning with gold chloride stabilises the silver, and thiosulphate removes excess silver remaining in the tissue. We all like to say "Reticulin" but "Reticulum" is the correct word. Michael Titford USA Pathology Mobile AL USA = ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Microarray info thanks!
Thank you everyone for all the information about microarray equipment. As usual on the Histonet, everyone was quick with the information, generous with their time, and informative. Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Microarray questions
We are thinking of making our own microarray blocks instead of purchasing someone elses slides. Can someone: 1) Recommend a punch to use? 2) Which size diameter tip is best? (for general use) 3) are the tips re-usable, or are they single use? I attended April Watanabes's workshop on microarray preparation at the Fort Lauderdale meeting but the equipment discussed in that workshop was Cadillac, where I need more "life in the trenches" type equipment. Thank you in advance. Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Coverslipping
Before the days of automatic coverslippers, I have seen coverslipping done in large quantities from above and from below. I guess it depends what you are comfortable with. In either event, you want to get a "squeegee" effect so no air bubbles are trapped in the mountant. You need more control when coverslipping thick sections, or whole mounts. With Canada balsem, there was less problem with drying back, but that is all history now. One technologist I saw at a London hospital would coverslip and then dip the whole slide in xylene and then wipe it off . It worked surprisingly well. Mike Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Seasons Greetings/Thank you Dr Margraf.
Seasons Greetings to everyone from the Heart of Dixie, and thank you Dr Linda Margraf and colleagues for hosting the "Histonet". I get to keep in touch and not have to leave my office! Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Slide Crusher
Debbie Boyd in Virginia asks about slide crushers: At the National Society meeting in Birmingham recently, a company called Creative Waste Solutions displayed a slide crusher. It grinds them up and the slide labels too until it is just like very roughly ground sugar. The instrument costs several thousand dollars, but, better still, you can rent it. They will rent you the instrument, and give you bags and boxes to put the ground up slides in. Then I guess it is off to the landfill. Their telephone number is (888) 795 8300 (www.slydeater.com). Jim Maynard, a histotechnologist, represents them in Florida. his number is (352) 978 9197 (I will need to rent one when I finally retire and I have to empty my office!) Mike Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fetal demise placenta
Maria asks about fetal demise placenta: We gross in all fetal demise placenta (usually four blocks).If the clinicians desire genetics, they take?samples from the fetus and the placenta up on the floor. (There is only a three day window for growing fibroblasts). The fetus gets a gross only treatment with weight, measurements and a photograph. If the clinicians or family desire an autopsy, whatever the gestational age, we have the parents sign an autopsy consent, as if for an adult. Regards Michael Titford USA Pathology Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] NSH Meeting in Alabama
With the NSH Annual Convention in Alabama this year, I don't want any corny jokes on the Histonet?about my adopted home state!? Might hurt my feelings! Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] CoPath computer question
We are about to start using the CoPath computer system from Sunquest. With previous systems we could obtain a surgical pathology accession number (for a frozen section for example) and put the demographic data and history in later. We have been told we cannot do that with CoPath. What do other labs with CoPath do? Thank you in advance Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] More Lab Week trivia
One year, everyone brought in photographs of their pets. There was a competition to guess which pets belonged to which lab employees. They say people look like their pets Mike Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] More about distilled water
I would like to add to the cooments of others concerning distilled water and deionised water: 1) I think most histology labs use deionised water as it is cheaper and easier? to obtain. 2) However, even deionised water will turn red with Schiff reagent. 3) For silver stain solutions, our lab uses "triple deionised" where the water passes through three tanks of resin, giving greater purity. 4) If your lab is required to be CAP complient, you will need monthly to check the resistivity and get a sample cultured.(It makes good sense even if you do not have to be CAP complient). 5) Deionised water systems can start growing Gram negative organisms in the tubing so you may want to put a filter over the actual spigot, and change that periodically. Mike Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Brain containers
im Wheelock asks about brain containers: We use the 5.7 liter polyethylene containers from Richard Allen (Cat # 5357). When fixing whole brains we suspend the brain in the formalin in a hair net. We keep the brain slices in these containers too. Michael Titford USA Pathology Mobile AL USA ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Christmas/Dr Margraf
Christmas greetings from the Heart of Dixie, and thanks to Dr Margraf,?her collegues and?her facility?for hosting the Histonet. I enjoy keeping up with histotechnology through the Histonet. Mike Titford USA Pathology Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Inspection of research labs / ISO 9000?
Kimberly Tuttle in Baltimore asks about lab inspections. I attended a workshop at the NSH S/C in Toronto where Diane Sterchi gave a really good workshop entitled, "Cracking the code: regulatory issues in the laboratory".? She spoke about the ISO 9000 certification requirements?that some research laboratories, I think drug company laboratories, must follow. Their requirements are really strict. Much more complicated than CAP.?If?Joe Nocito thinks he has problems now, he had better hope the CAP does not start ?following the ISO?guidelines! Michael Titford Pathology USA Mobile AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
