RE: [Histonet] Re: Cooling paraffin blocks with ice
Glad if that is some of the information you were hinting at. Joelle Weaver MAOM, HTL (ASCP) QIHC > From: carl.ho...@kcl.ac.uk > To: histonet@lists.utsouthwestern.edu > Date: Sun, 30 Sep 2012 17:27:49 + > Subject: [Histonet] Re: Cooling paraffin blocks with ice > > Nice one, Joelle. > > Good points > I thank you for that sophisticated paper! > > Yep, wax is not JUST wax. > It indeed has a crystalline, sophisticated structure. > Yes, cutting good sections depends upon temp/speed of cutting/angle of > knife/additives/effectiveness of processing. > May be more variables ...( eg: sharpness of blade, humidity) > ( Them crystals "slide" over each other, with heat.thus COLD tends to > minimize compression of section: ironically, so does heat: it momentarily > expands the crystals) > > I recall a paper by Edward Brain...in the 70sproposing the addition of > naphthalene to Pwax to improve penetration/cutting. > It worked very wellthen we realised that this additive was...NOT an > appropriate one;-) > Them were the days when we had Tea in the Lab andsmoked an occasional > fag, next to the dewaxing xylene ;-) > Interestingly, there have been no reports of xylene-related fires/cancers in > the ~ 100 years of xylene use. > Or, am I wrong ? > > > > > Respectfully, > > > Carl > > > > Carl Hobbs > Histology Manager > Wolfson CARD > School of Biomedical Sciences > Kings College London > Guys Campus > SE1 1UL > Tel: 020 78486813 > Fax: 020 78486816 > 020 78486813 > > > ___ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Cooling paraffin blocks with ice VS. Freezing Spray
I don't like ice-trays. Well, I tried them and.they swelled me blocks! Stick them blocks in the fridge, instead! Sure, cooling them blocks is damn fine to get very thin kidney sections... Diane! The wonderful Chief Tech of Gt Ormond St kids Hospital Where are you??? You know all this stuff abart getting THIN kidney sections from Pwax blocks. Try "hughing" on the block, before cutting? Like a kid's breath when they are sleeping. Ahhh. It's warm and, momentarily , expands them wax crystals? NB: Cooling them will have the same effect..weirdly Openly, Carl Caveat: each Lab MUST test out so many variables so as to appear Anal.. Fine...if you get damn fine results. However, share them results. Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Cooling paraffin blocks with ice
Nice one, Joelle. Good points I thank you for that sophisticated paper! Yep, wax is not JUST wax. It indeed has a crystalline, sophisticated structure. Yes, cutting good sections depends upon temp/speed of cutting/angle of knife/additives/effectiveness of processing. May be more variables ...( eg: sharpness of blade, humidity) ( Them crystals "slide" over each other, with heat.thus COLD tends to minimize compression of section: ironically, so does heat: it momentarily expands the crystals) I recall a paper by Edward Brain...in the 70sproposing the addition of naphthalene to Pwax to improve penetration/cutting. It worked very wellthen we realised that this additive was...NOT an appropriate one;-) Them were the days when we had Tea in the Lab andsmoked an occasional fag, next to the dewaxing xylene ;-) Interestingly, there have been no reports of xylene-related fires/cancers in the ~ 100 years of xylene use. Or, am I wrong ? Respectfully, Carl Carl Hobbs Histology Manager Wolfson CARD School of Biomedical Sciences Kings College London Guys Campus SE1 1UL Tel: 020 78486813 Fax: 020 78486816 020 78486813 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cooling paraffin blocks with ice VS. Freezing Spray
Well said Rene! In my 40 plus years as a supervisor, manager, technical support manager, and consultant; I have come across a broad spectrum of experienced and inexperienced histologists. Their knowledge base may be limited to their own experiences, or lack of it. A good supervisor or manager will be open to alternative methods of techniques. The mind is like a parachute; keep it open! Akemi Allison BS, HT(ASCP)HTL From: Rene J Buesa To: Jenny Vega ; joelle weaver ; "histonet@lists.utsouthwestern.edu" Sent: Sunday, September 30, 2012 7:17 AM Subject: Re: [Histonet] Cooling paraffin blocks with ice VS. Freezing Spray Jenny: There is a saying that "life is too short to drink cheap wine". In the same way life is too short to be frustrated daily working at a place where work is like a daily uphill battle. For what you describe you know your trade. Start looking for another place although do not expect that your ideas will always be well received. "Older, trained on the job, and with lots of experiences" supervisors usually are not very open to suggestions, especially when those ideas conflict with what they are used to do because they do not know the scientific basis of what they are doing. The less open to suggestions a person is, the more ignorant they are likely to be. René J. From: Jenny Vega To: joelle weaver ; histonet@lists.utsouthwestern.edu Sent: Saturday, September 29, 2012 6:26 PM Subject: Re: [Histonet] Cooling paraffin blocks with ice VS. Freezing Spray Thanks everybody for your answers. I cant respond them all but I concluded that the best way to get good sections is too chill the blocks on ice because I agree that it facilitates the process. I really don't understand why my supervisor depends so much on freezing sprays to cut and the pathologist has never complained about artifacts caused by them but I do believe that they are present because I have seen them getting formed. It makes sectioning difficult because you try to get sections free of holes and that contributes to the problem. At my lab is the same thing. My supervisor is in charge of the embedding and she just use the ice only for hardening the paraffin block. We don't have a standard embedding center with cold plate. Since is a small lab we just have a heating plate where we handle the specimens and place them in the molds and we cool them on ice trays. After they are removed from the molds they are placed on the counter in numerical order and they reach room temperature and get warm. I do think that if they get cold and moistened since the beginning it can facilitate the sectioning process except for certain tissues that are not well processed. In my lab I change the reagents in the tissue processor weekly but they get dirty too quickly because we processes a lot of breast and colon tissue so is hard to get perfectly processed tissue daily. We are under a tight budget and we can't waste materials too quickly. This situation with my supervisor has caused me a lot of frustration. I have noticed that every tech has their own method to do things but unfortunately there are people who are not receptive to new ideas and they immediately criticize and say you are wrong specially if you are young and you have recently started your career. I have though on several occasions that all I learned in histotech school have been worthless because everybody in the lab does things differently and this has made me question my ability of being a good tech because I have experienced difficulty using their microtomy technique but I have realized that I am not wrong. Another issue was the use of microtome blades. Since I started my supervisor has told me to use the minimum amount of blades as possible because we are under a tight budget but I have noticed that some of those blades are of poor quality and since we cut a lot of tissues that are hard, calcified or have sutures they wear the blades too quickly. It's hard to cut many blocks using only one blade that you use to trim and section. I have realized that I am successful obtaining good sections when I chill the blocks on ice for a couple of minutes and change the blade immediately after encountering difficulties with a block. It's not worth to sacrifice the quality of the samples just to be cheap and save some money. My supervisor is a great person and tech with a lot of experience but she is not very open to new ideas. Her demands to save money can be unrealistic when the correct technique is not being used . With her technique of course you are going to waste excess of freezing spray and blades but she doesn't understand this. Perhaps I need a change of environment which is unfortunate but is difficult to work like this. Thanks everyone for their input On Sat, Sep 29, 2012 at 8:56 AM, joelle weaver wrote: > Jenny > > My experience and training is to use some method involving ice o
[Histonet] freeze spray vs ic for microtomy There is a petient on the other side of the coverslip
&nb= sp;Perhaps you can tell your mgr that you respect her style and
method, and= show her that your style and method is just as effective.
When Lee Luna wa= s my manager he said time after time "techniiques
are as varied as technici= ans". If at the end of the day you are
putting out an acceptable number and= quality of slides then perhaps
your boss needs to accept your method as on= e way of doing things.
Regarding freeze sprays, never been a big fan, I sti= ll do not
believe there are safe for the ozone. Respect your mgrs differenc= es,
and prove to her you can be just as effective. I have been on both
side= s of this issue and have changed my techniques over the years
several times= when I discivered someone else had a better technique.
SOmetimes you have = to check that ego at the door and remember there
is a person on the other s= ide of the coverslip.
Nick(Rocky) Madary, HT/HTL(ASCP)QIHC
<= DIV style="MARGIN: 5px 0px; BORDER-TOP: #bcbcbc 1px solid">
On 09/29/12, [DEL: histonet-requ...@lists.utsouthwestern.edu :DEL]
wrote:
&nbs= p;
= Send Histonet mailing list submissions to
[1]histonet@lists.u= tsouthwestern.edu
To subscribe or unsubscribe via the World Wide= Web, visit
[2]http://lists.utsouthwestern.ed= u/mailman/listinfo/histonet
or, via email, send a message with subje= ct or body 'help' to
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You can reach the person managing the list at
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When r= eplying, please edit your Subject line so it is more specific
than "Re: = Contents of Histonet digest..."
Today's Topics:
1. Re: Pa= raffin and Tissue (Jackie O'Connor)
2. Caspase 8 for Mouse (Elizabeth Ca= meron)
3. Stainer for sale (Adrienne Anderson)
4. Re: Histonet Digest= , Vol 106, Issue 35 (Galina Deyneko)
5. Cooling paraffin blocks with ice= VS. Freezing Spray (Jenny Vega)
6. Re: Cooling paraffin blocks with ice= VS. Freezing Spray (C.D.G.)
7. Cooling paraffin blocks with ice VS. Fre= ezing Spray
(Contact HistoCare)
8. AW: [Histonet] Cooling paraffin bl= ocks with ice VS. Freezing
Spray (Gudrun Lang)
9. RE: Cooling paraffi= n blocks with ice VS. Freezing Spray
(joelle weaver)
10. Re: Cooling = paraffin blocks with ice VS. Freezing Spray
(Jackie O'Connor)
11. Re:= Cooling paraffin blocks with ice VS. Freezing Spray
(Rene J Buesa)
<= BR>
--
Message: 1
Date: Fri, 28 Sep 2012 13:05:22 -0400 (EDT)
From= : "Jackie O'Connor" <[5]b427...@aol.com>
Subject: Re: [Histonet] Para= ffin and Tissue
To: [6]deshsmi...@gmail.com, histonet@li= sts.utsouthwestern.edu
Message-ID:
<8CF6BB[7]294563e1e-1274-49...@webmail-m125.sysops.aol.com>
Content-T= ype: text/plain; charset="us-ascii"
It has been my experience = that tissues that remain in paraffin too
long (like over a weekend) become = brittle and hard. If we are
embedding over 300 blocks, those blocks may rem= ain in the embedding
station for up to 6 hours - but I personally strongly = recommend
sticking to your SOP for processing. Besides, if your SOP says pa
raffin for 3 hours, leaving them longer is a violation of your SOP.
Jack= ie O'
-Original Message-
From: Demetria Ross <= [8]deshsmi...@gmail.com>
To: histonet
Sent: Thu, Sep 27, 2012 5:21 pm
Subjec= t: [Histonet] Paraffin and Tissue
I'm curious to know how lo= ng can tissue stay on the machine in
paraffin
efore it becomes a problem= I have left tissue stay in paraffin 30
min-2
ours before I take it off = but not on a daily basis Thanks in advance
_= _
istonet mailing list
[9]isto...@lists.uts= outhwestern.edu
ttp://lists.utsouthwestern.edu/mailman/listinfo/hist= onet
--
Message: 2
Dat= e: Fri, 28 Sep 2012 17:59:57 +
From: Elizabeth Cameron <[10]El= izabeth.came...@jax.org>
Subject: [Histonet] Caspase 8 forMouse<=BR>To:
"[11]histonet@lists.utsouthwestern.edu"
<[12]histonet@lists.utsouthwestern.edu>
Message-ID:
Content-Type: text/plain; charset="us-ascii"
Anyone = out there know of a caspase 8 that works well on mouse
tissue?
Thanks!-Liz
The information in this email, including attachments, may= be
confidential and is intended solely for the addressee(s). If you
believ= e you received this email by mistake, please notify the sender
by return em= ail as soon as possible.
--
M= essage: 3
Date: Fri, 28 Sep 2012 15:26:04 -0400
From: Adrienne Anders= on <[14]rennie1...@yahoo.com>
Subject: [Histonet] Stainer for s= ale
To: [15]histonet@lists.utsouthwestern.edu
Message-I= D: <15FE6F08-1190-4A
Re: [Histonet] Cooling paraffin blocks with ice VS. Freezing Spray
Jenny: There is a saying that "life is too short to drink cheap wine". In the same way life is too short to be frustrated daily working at a place where work is like a daily uphill battle. For what you describe you know your trade. Start looking for another place although do not expect that your ideas will always be well received. "Older, trained on the job, and with lots of experiences" supervisors usually are not very open to suggestions, especially when those ideas conflict with what they are used to do because they do not know the scientific basis of what they are doing. The less open to suggestions a person is, the more ignorant they are likely to be. René J. From: Jenny Vega To: joelle weaver ; histonet@lists.utsouthwestern.edu Sent: Saturday, September 29, 2012 6:26 PM Subject: Re: [Histonet] Cooling paraffin blocks with ice VS. Freezing Spray Thanks everybody for your answers. I cant respond them all but I concluded that the best way to get good sections is too chill the blocks on ice because I agree that it facilitates the process. I really don't understand why my supervisor depends so much on freezing sprays to cut and the pathologist has never complained about artifacts caused by them but I do believe that they are present because I have seen them getting formed. It makes sectioning difficult because you try to get sections free of holes and that contributes to the problem. At my lab is the same thing. My supervisor is in charge of the embedding and she just use the ice only for hardening the paraffin block. We don't have a standard embedding center with cold plate. Since is a small lab we just have a heating plate where we handle the specimens and place them in the molds and we cool them on ice trays. After they are removed from the molds they are placed on the counter in numerical order and they reach room temperature and get warm. I do think that if they get cold and moistened since the beginning it can facilitate the sectioning process except for certain tissues that are not well processed. In my lab I change the reagents in the tissue processor weekly but they get dirty too quickly because we processes a lot of breast and colon tissue so is hard to get perfectly processed tissue daily. We are under a tight budget and we can't waste materials too quickly. This situation with my supervisor has caused me a lot of frustration. I have noticed that every tech has their own method to do things but unfortunately there are people who are not receptive to new ideas and they immediately criticize and say you are wrong specially if you are young and you have recently started your career. I have though on several occasions that all I learned in histotech school have been worthless because everybody in the lab does things differently and this has made me question my ability of being a good tech because I have experienced difficulty using their microtomy technique but I have realized that I am not wrong. Another issue was the use of microtome blades. Since I started my supervisor has told me to use the minimum amount of blades as possible because we are under a tight budget but I have noticed that some of those blades are of poor quality and since we cut a lot of tissues that are hard, calcified or have sutures they wear the blades too quickly. It's hard to cut many blocks using only one blade that you use to trim and section. I have realized that I am successful obtaining good sections when I chill the blocks on ice for a couple of minutes and change the blade immediately after encountering difficulties with a block. It's not worth to sacrifice the quality of the samples just to be cheap and save some money. My supervisor is a great person and tech with a lot of experience but she is not very open to new ideas. Her demands to save money can be unrealistic when the correct technique is not being used . With her technique of course you are going to waste excess of freezing spray and blades but she doesn't understand this. Perhaps I need a change of environment which is unfortunate but is difficult to work like this. Thanks everyone for their input On Sat, Sep 29, 2012 at 8:56 AM, joelle weaver wrote: > Jenny > > My experience and training is to use some method involving ice or at the > very least a cold-retaining tray made to chill blocks. I also was taught > this method in histology school , in clinical training at four quite > large institutions, and also have used some variation of an ice cooling > method in every instance in my career working in clinical and research > settings. There are always variations in technique from lab to lab, but > freezing spray has been generally discouraged for constant use at the > microtome, since it can introduce artifact in sections if over used. ( It > is pretty easy to see the effect on the face of the paraffin block, and > that is not even under the microscope.) I actually almost never use > freezing spray
Re: [Histonet] Negative Controls in IHC
Negative controls for IHC has been discussed before many times but it keeps pupping up every noew and then. This is my take on it: 1- if CAP requires them, so then do it. You do not want to have a negative issue in your inspection 2- it is always good to have them for quality control of your procedure and to prevent any legal issues down the road 3- if I have a case requiring several antibodies, I only run a negative control for the tissue series (the block in question) but not for every antibody but if in that antibodies series there are different detection systems, I run a negative control for each René J. From: Ann Specian To: histonet@lists.utsouthwestern.edu Sent: Saturday, September 29, 2012 1:56 PM Subject: [Histonet] Negative Controls in IHC I have a question in regard to eliminating the use of negative controls when using a non-avidin-biotin detection system. Do you not feel that negative controls may still need to be run in tissues which are likely to be pigmented such as lymph node, skin and liver? We were thinking to eliminate the use of negatives for other tissues (cervical, GI, etc.) only.What is the general consensus? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
