[Histonet] UC San Diego Anatomic Pathology Histology Labs is Hiring

[Va Paula Sicurello via Histonet]

Hello,
We are hiring a histotechnologist for our growing histology lab.  UCSD is 
adding a new hospital and soon our volume will increase.  We are looking for an 
experienced histotech.  Please go to: 
https://jobs.ucsd.edu/bulletin/job.aspx?cat=health=post_in=81586. 
Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
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Re: [Histonet] Start up Vet/Animal histology laboratory

[Va Paula Sicurello via Histonet]

Hi Linda,
I've set up three histology labs for animal research: one in an academic 
setting, one for a pharmaceutical firm, and one for a research foundation.  
1. Having money to buy the proper equipment is tricky, start with used 
equipment and make due until you can afford to buy new.  As long as you buy the 
equipment from somewhere that       either provides a warranty or the equipment 
is so inexpensive that you don't care if it breaks (as long as it gets you 
going).
2. Having the investors (or whom ever is supplying the majority of the funding) 
understand that often times histology labs that are providing a service to 
researchers do not make money.         In the early stages they are black pits 
you pour money into.
3. Building up a clientele requires a lot of advertising and offering low 
prices (undercutting the competition even if it means you suffer a loss).
4. Have a method of billing, invoicing, and collecting on those invoices can be 
a challenge.  Simple bookkeeping software like Quickbooks or FileMaker will 
help you considerably.  
5. Write your SOPs as soon as you can.  Having the methodology of every 
technique, stain and procedure you will be doing is invaluable.  When you get 
around to be able to hire staff           these will be a lifesaver.  Having 
established SOPs allow you to make sure your staff also follow the same 
procedure.  Reproducability and consistency is a must when establishing a       
  histology lab.  There is no shame in borrowing heavily from SOPs you have 
come across in the past.  Rely on colleagues to provide SOPs and pointers.
6.  Make sure that the place you select for the business allows for proper 
ventilation of all the reagents and chemicals.  You want to limit your exposure 
to the chemicals in the lab to the          maximum point possible.  Safety 
should be of the utmost importance in the new lab.
Those are just a few things I can think of.  Most important is to enjoy the 
ride-it's going to be hard and sometimes frustrating work-but building the 
business is part of the fun. Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
 

On Saturday, July 9, 2016 5:08 PM, Linda via Histonet 
 wrote:
 

  Hello Histoland,
I am inquiring if anyone has started their own lab to do histology services for 
veterinarians or animal research tissues?
Or anyone who has started a private lab?
What are your experiences?  What didn't you expect?  
Thank you in advance.
Regards,
Linda Dee, BGS,HT(ASCP) lmd...@yahoo.com

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[Histonet] 10% NBF Question

[Va Paula Sicurello via Histonet]

Hello Listers,

Has anyone had any problems with fixation using Richard-Allan Scientific 10% 
NBF?  Some of our gastric biopsies are looking a little off.  The rest of the 
biopsies fixed in the same NBF and processed on the same processor look fine.

This issue would have come up in the last couple of weeks.

I know that Richard-Allan recalled their NBF a couple of years ago because the 
% was 0-3 not 10%.

Please let me know if any of you are experiencing something similar.

Sincerely,


Paula


Paula Sicurello
Histotechnology Specialist
UC San Diego Health


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[Histonet] Results of My Histology Survey-Finally!

[Va Paula Sicurello via Histonet]

Well Listers,
I finally get around to posting my survey.  Only 35 people took it but it still 
provided useful information and I got an A in my class.
Thank You to the 35 who participated
All the data:             23 HT's, 8 HTL's, 4 with other certification (2 with 
HT/HTL) took my survey
             24 years of experience average.               Range 5 
to 45 years.             49 blocks per hour, average cutting.           
  Range 19 to 120             69 slides per hour, average.                      
Range  35 to 160/hour             7 average number of staff.        
                 Range 1 to 22
             278 blocks per day per lab, average.         Range 10 
to 1000
             467 H slides per day, per lab, average.  Range 10 to 
1500
              92 IHC slides per day, per lab, average      Range 2 
to 300
             19 special stain slides per day, per lab, average     Range 1 to 
100

 My take away from this was that we are getting older, we are working faster 
with fewer staff and we use automation to our advantage.
I think none of this is surprising to you.    


Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
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[Histonet] Traveling Histologists?

[Va Paula Sicurello via Histonet]

Good Morning Listers,
Are there companies out there that work with traveling histologists?  If so, 
can you send me their contact information?
Companies dealing with traveling histologists are free to contact me as well. 
Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
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[Histonet] Workload and Productivity Survey closing this week

[Va Paula Sicurello via Histonet]

Good Morning Listers,
I will be closing my survey this week on Friday.  If anyone else would like to 
take it, please follow this link:
Histology Questionaire Survey

|   |
|   |   |   |   |   |
| Histology Questionaire SurveyWeb survey powered by SurveyMonkey.com. Create 
your own online survey now with SurveyMonkey's expert certified FREE templates. 
|
|  |
| View on www.surveymonkey.com | Preview by Yahoo |
|  |
|   |


 Thank you for helping me with my project.  
Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
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[Histonet] Histology Questions part 2

[Va Paula Sicurello via Histonet]

Good Evening Netters,
I've created a short survey using Survey Monkey.  Please take my survey and 
help me gather information I can use in my class and to help update the 
productivity standards and staffing benchmarks for our Histology labs.
https://www.surveymonkey.com/r/8HFSY3V

Thank you and thanks to those who took my first survey.  The results look 
interesting.

Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
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[Histonet] Histology Lab Survey Questions

[Va Paula Sicurello via Histonet]

Hello Netters,
Thank you for taking my survey.  I will use your replies for my research 
project and I will also let everyone on the list know the results as well.
From what I've found, the surveys performed in the past asked similar questions 
but more at the institution level.  I would like to get information at the 
individual level.  There are 16 questions.
Please reply to this email offline with your answers.
Thank you for helping me.  Let's hope I get an A! :-)
Here we go:Questions aboutyou:

1. Education (highest achieved):

a.  High school

b. Associates degree

c.  Bachelor’s degree

d. Master’s degree

e.  PhD or MD

2. Certification:

a.  None

b. HT

c.  HTL

d. Other, please describe

3. How did you learn histology?

a.  On the job

b. Histotechnology school

4. How many years of experience in histology?

5. How many blocks can you embed in an hour?

6. How many of the following can you produce in an 8 hourday?   
    (You can calculate from howmany of each task 
you can perform in one hour: 20 blocks with one slide each per hour = 160 
blocks and 160 slides)

a.  Blocks

b. Slides


Questions aboutyour place of work:

7. How many blocks does your lab embed a day?

8. How many staff does your lab have?

a.  Technical (HT, HTL, etc.)

b. Non-technical (lab assistants, etc.)

9. How is IHC handled in your lab?  

a. Separate staff (if so how many?) 

b. All staff perform

10.  Howmany IHC slides does your lab stain per day?

11.  Howmany of the following can your lab run?

a.  Antibodies (total number available for testing)

b. Special Stains (total number available for testing)

12.  Doesyour lab use automation?  Yes or No

Ifyes, please describe:

13.  Whoprepares the slides prior to release to the pathologist?

14.  Whofiles the blocks and slides?

15.  Whattype of institution do you work at?

a.  Hospital based Histology lab

b. Clinic based Histology lab

c.  Independent lab (such as a referral lab)

16.  BonusQuestion:  Has your family stopped askingwhy you come home with blue 
or pink-orange fingers?


OPTIONAL: Please add your comments, suggestions,or thoughts.
 Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
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[Histonet] Histology Related Questions?

[Va Paula Sicurello via Histonet]

Hello Netters,
I am taking a research techniques class for my MBA this term and need to have a 
project that I can ask quantifiable questions.  I was thinking about 
productivity and benchmarks in the Histology and IHC labs.
If I send an email with my questions, will you be kind and answer them in a 
reply email?
I hope to keep it between 15-20 questions.
I even promise to share my results once I compile all the information.
Suggested questions are also gratefully accepted.
Toodles! Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
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[Histonet] Test email

[Va Paula Sicurello via Histonet]

Good Morning Netters,

This is a test to see if I can get an email through using this email account.

My posts haven't been making it through and I know you miss me. 
 Sincerely,

Paula

Paula Sicurello
Histotechnology Specialist
UC San Diego Health

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[Histonet] Histology Related Questions?

[Va Paula Sicurello via Histonet]

Hello Netters,
I am taking a research techniques class for my MBA this term and need to have a 
project that I can ask quantifiable questions.  I was thinking about 
productivity and benchmarks in the Histology and IHC labs.
If I send an email with my questions, will you be kind and answer them in a 
reply email?
I hope to keep it between 15-20 questions.
I even promise to share my results once I compile all the information.
Suggested questions are also gratefully accepted.
Toodles! Sincerely,
Paula
Paula SicurelloHistotechnology SpecialistUC San Diego Health
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[Histonet] BSL-2 question

[Va Paula Sicurello]

Hello Everybody out there in virus land,

I am getting a room ready for BSL-2 level virus imaging.  I can't get any 
information regarding how safe the cells are once the virus has been 
transfected (?) in to them.

I have a BSL-2 level bio-hood, I have a room with a door and I have a brand, 
spanking new deconvolution with FRET and an incubator for live cell imaging.

I need to know if the people in the room are safe from the virus once it's 
inside the cells.  I can't get the air pressure changed in the room so I need 
to know what other precautions we might need to take.

Thanks to all who can give me any pertinent information.

Paula  :-)

 Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core for Micro Imaging(C-MI)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


C-MI for your imaging needs.



  

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Re: [Histonet] RE: OT: Early Friday Hour of Fuming

[Va Paula Sicurello]

I also feel your pain.  I have lab where I do electron microscopy (clinical {if 
they'd send me any} and research), confocal microscopy and histology.  I also 
have added cryosectioning and will be adding a micro-CT and a deconvolution 
microscope (working at a BSL-2 level) with no increase in hours.  I'm 80% and 
my lab will be shut down at the end of the year because I do not bring in 
enough revenue.

Clinical or research-it appears there aren't enough of us and we aren't 
appreciated.  Have the CEO of your company send out a blanket e-mail mentioning 
a reduction in staffing when everyone knows that I'm the only one, talk about 
lack of job security.

Grrr,  hey at least the sun is shining and there are blue skies!

Paula  :-)


Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core for Micro Imaging(C-MI)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

C-MI for your imaging needs.


--- On Mon, 9/21/09, Roberta Horner r...@psu.edu wrote:

 From: Roberta Horner r...@psu.edu
 Subject: [Histonet] RE: OT: Early Friday Hour of Fuming
 To: Breeden, Sara sbree...@nmda.nmsu.edu, 
 histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
 Date: Monday, September 21, 2009, 7:36 PM
 That's alright whenever I take
 vacation I come back to all the work sitting here waiting
 for me.  I just took 2 weeks off and it took 3 weeks to
 catch up so I feel your pain.
 Roberta Horner
 Penn State University
 Animal Diagnostic Lab
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu]
 On Behalf Of Breeden, Sara
 Sent: Monday, September 21, 2009 2:23 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] OT: Early Friday Hour of Fuming
 
 I have just found out this morning that I will be unable to
 attend NSH
 this year.  This alone is unfortunate news for me but,
 interestingly
 enough, is not due to budget issues.  The reason I am
 fuming this
 morning is not because there is no funding for my
 attendance (that was
 already in place), but because there is such a serious
 shortage of
 histotechs in this country that there is no one to cover
 me!  A
 traveler is out of the question (that's a budget issue!);
 anyone that
 might be hiding and is not known to me to be available in
 our city would
 have to have about a month's lead time to get vetted for
 the position
 through the State.  Anyone that thinks that histology
 is not important
 should try to fill a position when there is such a shortage
 of trained
 and qualified technicians.   If we, as
 histologists, do not begin to
 make ourselves more visible and promote our profession as a
 worthy,
 fulfilling and job-secure life, more and more of us will
 find ourselves
 in this position. Thank you - I feel better for having
 Fumed.  There is
 no need to reply as I know many of us are in the same
 boat.  I was in
 the boat and lost my paddle!
 
  
 
 Sally Breeden, HT(ASCP)
 
 NM Dept. of Agriculture
 
 Veterinary Diagnostic Services
 
 PO Box 4700
 
 Albuquerque, NM  87106
 
 505-841-2576
 
  
 
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Re: [Histonet] EM processing

[Va Paula Sicurello]

I have never heard of an EM lab making their own resin for embedding.  We 
almost all buy the resin kits or the separate components to make either a Epon 
812 equivalent or Spurrs.

The amount of time it takes to process is usually tissue dependent.  On 
average, it takes one day to process (if the tissue is 1mm cubed or less) and 2 
days in the oven for the resin to polymerize.  You can speed things up if you 
microwave process.

If you have any questions, please feel free to contact me.

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core for Micro Imaging(C-MI)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

C-MI for your imaging needs.


--- On Sat, 8/29/09, histopa...@aol.com histopa...@aol.com wrote:

 From: histopa...@aol.com histopa...@aol.com
 Subject: [Histonet] EM processing
 To: histonet@lists.utsouthwestern.edu
 Date: Saturday, August 29, 2009, 3:36 AM
 Does your lab make their own plastic
 resin to process and embed specimens or do you buy it
 commercially?
 What is the average time it take to process a specimen?
 
 Thanks in advance to all who respond.
 P.Eneff
 OU Medical Center
 Oklahoma City, OK
 
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Re: [Histonet] Organizing of cassettes for processing

[Va Paula Sicurello]

When I batted clean up and loaded the processors, the residents grossed in the 
specimens and placed them in a basket in a container of formalin with a lid.  
The grossing was done and the containers were kept on the downdraft table.

When it came time to load the processors I put the cassettes in numerical order 
and in separate baskets (smalls like GI biopsies and needle biopsies went 
together in one basket, fatty samples like breast tissue in another basket, 
etc).  I did this on the downdraft table and placed the cassettes in numerical 
order.

This helped us to find out if samples were mislabelled (2 cassettes with the 
same number) or missing (samples labelled A-F and D was missing).  This saved 
our bacon many times since I would make notes or ask the resident where the 
sample was or which was which.

It also made it easier for the embedders since I placed the notes regarding the 
missing cassettes by the embedding stations.  Plus they could embed the block 
in numeric order so the slides that needed to get out first were cut and 
stained first.

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core for Micro Imaging(C-MI)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

C-MI for your imaging needs.


--- On Wed, 8/26/09, ba...@gundluth.org ba...@gundluth.org wrote:

 From: ba...@gundluth.org ba...@gundluth.org
 Subject: [Histonet] Organizing of cassettes for processing
 To: histonet@lists.utsouthwestern.edu
 Date: Wednesday, August 26, 2009, 8:38 PM
 
 Hi all -
 
 For those that do batch overnight processing, how do you
 organize the
 cassettes?
 
 Currently we have 2 path assistants that gross throughout
 the day, and each
 puts their cassettes as they are grossed into a bucket of
 formalin.  At the
 end of the day a histotech drains the formalin off, rinses
 the cassettes in
 water, then manually puts the cassettes into order
 according to our
 worklist, with rush cases being put up front.  The
 baskets are then loaded
 onto the tissue processor (Sakura VIP5 and VIP6).
 
 We are wondering if there are some other ideas of how to
 streamline this
 process.  One thought was to have the cassettes
 loaded/organized into the
 tissue processing basket as they are grossed, but have a
 concern about
 formalin exposure while doing this.
 
  Any thoughts would be greatly appreciated.
 
 Barb Moe
 Gundersen Lutheran Medical Center
 La Crosse WI
 
 
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Re: [Histonet] water on slides?

[Va Paula Sicurello]

I'm getting the same eosin bleeding and water on the slides.  I'm using Hemo-D 
and CitriSolv.  I have changed the reagents and am still getting the eosin 
bleeding, the water on the slide is a new phenomenon.

Any suggestions?


Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core for Micro Imaging(C-MI)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

C-MI for your imaging needs.


--- On Mon, 8/24/09, Angela Bitting akbitt...@geisinger.edu wrote:

 From: Angela Bitting akbitt...@geisinger.edu
 Subject: Re: [Histonet] water on slides?
 To: histonet@lists.utsouthwestern.edu, Tina J. Hayes tina.ha...@va.gov
 Date: Monday, August 24, 2009, 6:00 PM
 We are having a very similar problem
 too. We change our alcohols and Histoclear like mad, but are
 having eosin bleeding out of the sections. I'm starting to
 wonder if it is the mounting media on our glass
 coverslipper. I ordered new bottle gaskets, so I guess I'll
 see what happens then.
 
 Angela Bitting, HT(ASCP)
 Technical Specialist, Histology
 Geisinger Medical Center 
 100 N Academy Ave. MC 23-00
 Danville, PA 17822
 phone  570-214-9634
 fax  570-271-5916 
  
 No trees were hurt in the sending of this email
 However many electrons were severly inconvienienced!
 
 
  Hayes, Tina J. tina.ha...@va.gov
 8/24/2009 12:41 PM 
 We are having a problem with what seems to be water on our
 slides.  We
 have had very high humidity levels in the air.  It
 seems that we look at
 the slides before taking them to the pathology office, one
 pathologist
 reviews them and sees little or no water droplets, but as
 they sit and
 another pathologist reads them later, like the following
 day, the
 droplets are very apparent.
 
  
 
 We use Clearite-3.  We have no xylene in our
 lab.  We also coverslip
 with Permount.
 
  
 
 Is it possible that the clearite-3 and/or Permount is
 absorbing moisture
 from the atmosphere to the slides?
 
 And if so, do you have any suggestions for combating this
 issue?
 
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 IMPORTANT WARNING: The information in this message (and the
 documents attached to it, if any) is confidential and may be
 legally privileged. It is intended solely for the addressee.
 Access to this message by anyone else is unauthorized. If
 you are not the intended recipient, any disclosure, copying,
 distribution or any action taken, or omitted to be taken, in
 reliance on it is prohibited and may be unlawful. If you
 have received this message in error, please delete all
 electronic copies of this message (and the documents
 attached to it, if any), destroy any hard copies you may
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RE: [Histonet] Cleaning oil off objectives

[Va Paula Sicurello]

I use a dog chewed old stick.

On another note,  how to clean off dried mounting media.  I have several 
objectives where some goombahs dragged the objectives through wet mounting 
media.

If y'all have any suggestions about removing dried mounting media.  Send them 
my way.  

4 of my 5 objectives are no longer objective after having their lenses clouded 
by the media.  ;-)


Paula 

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core for Micro Imaging(C-MI)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

C-MI for your imaging needs.


--- On Fri, 8/21/09, Pamela Marcum mucra...@comcast.net wrote:

 From: Pamela Marcum mucra...@comcast.net
 Subject: RE: [Histonet] Cleaning oil off objectives
 To: 'Rittman, Barry R' barry.r.ritt...@uth.tmc.edu, 
 histonet@lists.utsouthwestern.edu
 Date: Friday, August 21, 2009, 12:43 PM
 Really like the taser idea!  I
 could have used that for several people over
 the years.  
 
 Pam Marcum
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu]
 On Behalf Of Rittman,
 Barry R
 Sent: Thursday, August 20, 2009 10:36 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] Cleaning oil off objectives
 
 Adam
 Hi
 I would strongly recommend that the best approach is to
 train the people
 using the microscope, everyone is trainable although for
 some this may be a
 long learning curve.
 The use of a taser with the later individuals is strongly
 recommended!
 
 Several years ago the Zeiss representative in Iowa used the
 expanded plastic
 packing beads to wipe off the excess oil as he said this
 was much more
 absorbent for oil that lens tissue.
 We have also seen the use of soft wood tips with oil that
 is encrusted on,
 on the understanding that the wood is much softer than the
 lens.. Never
 completely happy with that concept.
 A lot depends on the type of lens that is being used.
 Some lenses, especially older ones may have a coating that
 is easily damaged
 even by Q tips.
 I would use lens paper first (don't be cheap skate with the
 lens tissue)
 then repeat using  a small amount of lens cleaner. The
 most difficult and
 usually the most contaminated seem to be the 40 due to its
 working distance.
 Most of the lens cleaners have isopropyl alcohol and some
 acetone. 
 If it really does not get all the oil after repeating a
 couple of times then
 can use acetone but don't flood the lens just use small
 amounts and wipe
 across the face. Follow this with lens cleaner and lens
 paper.
 Has always worked for me. This sounds a lengthy procedure
 but only takes a
 couple of minutes.
 Hope that this helps
 Barry
 
 
 
 From: histonet-boun...@lists.utsouthwestern.edu
 [histonet-boun...@lists.utsouthwestern.edu]
 On Behalf Of Adam .
 [anonwu...@gmail.com]
 Sent: Thursday, August 20, 2009 7:19 PM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Cleaning oil off objectives
 
 Hi all,
 
 You guys were so helpful on my last question, I'll ask
 another. We have a
 microscope shared by the floor with several objectives, and
 it's pretty
 common for the non-immersion objectives to get contaminated
 with oil. I
 asked the guy who is responsible for the scope about this.
 He said that they
 call someone from some company who carefully cleans the
 objectives with
 acetone and a Q-tip, which if done right works wonders but
 if done wrong it
 can damage the lenses. But he mentioned that the lenses are
 usually
 re-contaminated within a few weeks since so many people use
 the scope, so
 it's sort of a pointless endeavor. This system seems pretty
 silly to me... I
 feel like there must be an easier and cheaper way to clean
 the lenses
 without damaging them; I certainly don't want to be
 responsible for damaging
 a microscope that costs more than my yearly salary. What do
 you recommend?
 
 Thanks,
 Adam
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[Histonet] Core unit support

[Va Paula Sicurello]

Hello Everybody,

I was wondering what type of institutional support y'all have at your various 
institutions.  I work for a research foundation and they want to drop my salary 
and have the researchers grants pay my salary and almost everything else.  I am 
a fee-for-service lab and the CEO wants to see $25K in recharges between now 
and December.

We offer TEM, confocal, histology and cryosectioning.  We will be adding 
micro-CT and a deconvolution microscope that does FRET and can handle samples 
that have been transfected(?) with viral vectors.

Is she being unrealistic?  I am only 80% time and the only person down here.  
Should I start looking for a new position before she shuts me down?  How many 
people have had their facilities closed by the people who don't see the big 
picture?

Please let me know your experiences.  

Thanks,

Paula  :-}

Paula Sicurello

VA Medical Center San Diego

Veterans Medical Research Foundation (VMRF) 

Core for Micro Imaging(C-MI)

3350 La Jolla Village Dr., MC151

San Diego, CA 92161

858-552-8585 x2397



C-MI for your imaging needs.



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[Histonet] Support for core units

[Va Paula Sicurello]

Hello Everybody,

I was wondering what type of institutional support y'all have at your various 
institutions.  I work for a research foundation and they want to drop my salary 
and have the researchers grants pay my salary and almost everything else.  I am 
a fee-for-service lab and the CEO wants to see $25K in recharges between now 
and December.  

We offer TEM, confocal, histology and cryosectioning.  We will be adding 
micro-CT and a deconvolution microscope that does FRET and can handle samples 
that have been transfected(?) with viral vectors.

Is she being unrealistic?  I am only 80% time and the only person down here.  
Should I start looking for a new position before she shuts me down?  How many 
people have had their facilities closed by the people who don't see the big 
picture?

Please let me know your experiences. 

Thanks,

Paula  :-}


Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core for Micro Imaging(C-MI)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

C-MI for your imaging needs.


  

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Re: [Histonet] Clear-Rite 3

[Va Paula Sicurello]

Hi Rene,

Why do you not like the d-limonene solvents? 

I work with them and the results seem fine.  I have to mention that I use an 
AutoTechnicon Duo and cannot place it in a fume hood to vent the xylene 
vapors.  Without proper ventilation the safety people willnot allow the use of 
Xylenen.  I work in the research unit of a VA hospital and they will not 
provide larger fume hoods nor buy me a self contained processing sytem.

Please let me know what other safe alternatives (ones that can be used out on 
the bench) are you suggest.

Thanks,

Paula Sicurello

VA Medical Center San Diego

Veterans Medical Research Foundation (VMRF) 

Core for Micro Imaging(C-MI)

3350 La Jolla Village Dr., MC151

San Diego, CA 92161

858-552-8585 x2397



C-MI for your imaging needs.

--- On Wed, 8/5/09, Rene J Buesa rjbu...@yahoo.com wrote:

From: Rene J Buesa rjbu...@yahoo.com
Subject: Re: [Histonet] Clear-Rite 3
To: Histonet@lists.utsouthwestern.edu, BillO'Donnell 
billodonn...@catholichealth.net
Date: Wednesday, August 5, 2009, 2:01 PM

Bill:
Xylene substitutes are not all pretty much the same. There are d-Limonene 
derivatives (something to stay absolutely away from) and alkane derivatives 
with many known and unknown constituents.
Since you are about to change and not have any preference, your best option 
both in processing quality and price, would be to get a reliable supplier of 
mineral spirits (the same you can find at any home improvement store) and use 
it.
René J. 

--- On Wed, 8/5/09, O'Donnell, Bill billodonn...@catholichealth.net wrote:


From: O'Donnell, Bill billodonn...@catholichealth.net
Subject: [Histonet] Clear-Rite 3
To: Histonet@lists.utsouthwestern.edu
Date: Wednesday, August 5, 2009, 9:11 AM


Greetings!

We have been using Clear-Rite 3 here at our lab, and we are happy with
the product. Our supplier says it will be on back-order for some time
now. Our crack supply folks are looking for another source. I'm taking
another route to find out what products out there are comparable.

Are all Xylene Substitutes pretty much the same and there for pretty
much interchangable?
Are there some to stay away from?

Any help is appriciated.

William (Bill) O'Donnell, HT (ASCP) QIHC 
Lead Histologist
Good Samaritan Hospital
10 East 31st Street
Kearney, NE 68847 
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RE: [Histonet] Start Up Lab

[Va Paula Sicurello]

In the 80's I worked in a drug screening lab for the Navy and one step in the 
extraction involved fuming HCl (meaning pure HCl).  Several of the techs got 
nose bleeds every time they used the stuff.  I finally pried open a window 
(they were painted shut) and we turned on a fan the size of a propeller to blow 
the fumes out the window.

We used Xylene in a room with no ventilation and found out that high 
concentrations of xylene cause transient euphoria.  No wonder we thought 
every thing was funny during those days.

Safety people, they take all the fun out of research!  ;-)

Paula Sicurello

VA Medical Center San Diego

Veterans Medical Research Foundation (VMRF) 

Core Research Imaging Center (CRIC)

3350 La Jolla Village Dr., MC151

San Diego, CA 92161

858-552-8585 x2397



Your images flow through our CRIC.

--- On Thu, 7/23/09, Lynette Pavelich lpave...@hurleymc.com wrote:

From: Lynette Pavelich lpave...@hurleymc.com
Subject: RE: [Histonet] Start Up Lab
To: cb...@lexclin.com, montina.vanme...@pbrc.edu, theci...@yahoo.com
Cc: histonet@lists.utsouthwestern.edu
Date: Thursday, July 23, 2009, 3:53 PM

Ah, yes.  The '70's.  Our lab had 2 windows, both on the same side of
the wall.  The pathologists cut under one of the windows and it had one
of those box fans, pointing out.  In Michigan.  No hoods, an old
duo-autotechnicon and xylene in full use with disposal down the drain. 
Every day, I would go home not knowing if I went thru a red light or
not, arriving home , head still cloudy for about 30 minutes until it
cleared.  Ah..those were the days??  Although I miss it, I'm glad I had
to give up my coffee and donut when cutting.  Treats in the drawer are
gone, mouth pipetting is gone (not mine, but yes, the mouth does turn
black from silver nitrate!!)  Progress is good.  But I still like my
'70's music!!!  Bob Seger and old motown!!! ;)

 Montina Van Meter montina.vanme...@pbrc.edu 07/23/09 11:31 AM

I can remember back in the late 70's sitting at my microtome in a
clinical lab (that was the size of an elevator shaft = no
ventilation),where my supervisor would puff away on  cigarettes with an
ashtray perched on top of her microtome.  When we had inspections
everyone would put their contraband in their personal drawer.  I also
interned in an ENT lab that processed with celloidin (=ETHER) and didn't
have windows or a hood.  That supervisor also smoked in the lab!  I said
a prayer every day that I wouldn't blow up or be asphyxiated during my
two week stay.  


 

Montina J. Van Meter, HT (ASCP)
Lab Manager
Autonomic Neuroscience
Pennington Biomedical Research Center
6400 Perkins Rd.
Baton Rouge, LA  70791
225-763-2564

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Carol
Bryant
Sent: Thursday, July 23, 2009 9:04 AM
To: theci...@yahoo.com; Lynette Pavelich
Cc: Histonet
Subject: RE: [Histonet] Start Up Lab

Love it!  Too funny! 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
theci...@yahoo.com
Sent: Wednesday, July 22, 2009 6:59 PM
To: Lynette Pavelich
Cc: Histonet
Subject: Re: [Histonet] Start Up Lab

Dang maybe I should stop keeping my lunch in the cryostat. The fresh
unfixed tissue adds a certain je ne sais quoi to the flavor. :/
Sent from my Verizon Wireless BlackBerry

-Original Message-
From: Lynette Pavelich lpave...@hurleymc.com

Date: Wed, 22 Jul 2009 18:07:07 
To: lei...@buffalo.edu; histonet@lists.utsouthwestern.edu;
cing...@uwhealth.org
Subject: RE: [Histonet] Start Up Lab


Gosh.I remember the days sipping on my coffee and nibbling on a
fresh donut as I cut my morning slides!  Sigh..

 Merced M Leiker lei...@buffalo.edu 07/22/09 5:00 PM 
(lol some labs have a bench area as well as a desk area where food is 
allowed.)

--On Wednesday, July 22, 2009 3:19 PM -0500 Ingles Claire 
cing...@uwhealth.org wrote:

 In the lab?!? For shame. :)

 

 From: histonet-boun...@lists.utsouthwestern.edu on behalf of Cindy
DuBois
 Sent: Wed 7/22/2009 10:29 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Start Up Lab



 And a coffee pot.

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Merced M Leiker
Research Technician II
Cardiovascular Medicine
348 Biomedical Research Building
State University of New York at Buffalo
3435 Main St, Buffalo, NY 14214  USA
lei...@buffalo.edu
716-829-6118 (Ph)
716-829-2665 (Fx)

No trees were harmed in the sending of this email.
However, many electrons were severely inconvenienced.


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RE: [Histonet] Start Up Lab

[Va Paula Sicurello]

You might still have your fingers, but are your fingerprints still the same?  

Paula Sicurello

VA Medical Center San Diego

Veterans Medical Research Foundation (VMRF) 

Core Research Imaging Center (CRIC)

3350 La Jolla Village Dr., MC151

San Diego, CA 92161

858-552-8585 x2397



Your images flow through our CRIC.

--- On Thu, 7/23/09, Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov wrote:

From: Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov
Subject: RE: [Histonet] Start Up Lab
To: Smith, Allen asm...@mail.barry.edu
Cc: Histonet@lists.utsouthwestern.edu
Date: Thursday, July 23, 2009, 4:32 PM

Thanks to spell-check..it self-corrected me..incorrectly!
Thanks for setting it right.



-Original Message-
From: Smith, Allen [mailto:asm...@mail.barry.edu] 
Sent: Thursday, July 23, 2009 12:26 PM
To: Bartlett, Jeanine (CDC/CCID/NCZVED)
Cc: 'Histonet@lists.utsouthwestern.edu'
Subject: RE: [Histonet] Start Up Lab

I can't imagine why any one would use dioxin, which used to be used as
an insulator and heat convector in electron microscope transformers, as
a clearing agent.  I think you mean dioxane.


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Bartlett, Jeanine (CDC/CCID/NCZVED)
Sent: Thursday, July 23, 2009 12:13 PM
To: Edwards, R.E.; Montina Van Meter; Carol Bryant; theci...@yahoo.com;
Lynette Pavelich
Cc: Histonet
Subject: RE: [Histonet] Start Up Lab

Hah!  Sure do. 




-Original Message-
From: Edwards, R.E. [mailto:r...@leicester.ac.uk]
Sent: Thursday, July 23, 2009 11:51 AM
To: Bartlett, Jeanine (CDC/CCID/NCZVED); Montina Van Meter; Carol
Bryant; theci...@yahoo.com; Lynette Pavelich
Cc: Histonet
Subject: RE: [Histonet] Start Up Lab

You still  have  fingers!?.

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
Bartlett, Jeanine (CDC/CCID/NCZVED)
Sent: 23 July 2009 16:44
To: Montina Van Meter; Carol Bryant; theci...@yahoo.com; Lynette
Pavelich
Cc: Histonet
Subject: RE: [Histonet] Start Up Lab

And I used to use dioxin in my tissue processor and would actually dip
gauze in it to clean paraffin off my fingers. 


Jeanine Bartlett
Infectious Diseases Pathology Branch
(404) 639-3590
jeanine.bartl...@cdc.hhs.gov


-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Montina
Van Meter
Sent: Thursday, July 23, 2009 11:31 AM
To: Carol Bryant; theci...@yahoo.com; Lynette Pavelich
Cc: Histonet
Subject: RE: [Histonet] Start Up Lab

I can remember back in the late 70's sitting at my microtome in a
clinical lab (that was the size of an elevator shaft = no
ventilation),where my supervisor would puff away on  cigarettes with an
ashtray perched on top of her microtome.  When we had inspections
everyone would put their contraband in their personal drawer.  I also
interned in an ENT lab that processed with celloidin (=ETHER) and didn't
have windows or a hood.  That supervisor also smoked in the lab!  I said
a prayer every day that I wouldn't blow up or be asphyxiated during my
two week stay.  


 

Montina J. Van Meter, HT (ASCP)
Lab Manager
Autonomic Neuroscience
Pennington Biomedical Research Center
6400 Perkins Rd.
Baton Rouge, LA  70791
225-763-2564

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Carol
Bryant
Sent: Thursday, July 23, 2009 9:04 AM
To: theci...@yahoo.com; Lynette Pavelich
Cc: Histonet
Subject: RE: [Histonet] Start Up Lab

Love it!  Too funny! 

-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
theci...@yahoo.com
Sent: Wednesday, July 22, 2009 6:59 PM
To: Lynette Pavelich
Cc: Histonet
Subject: Re: [Histonet] Start Up Lab

Dang maybe I should stop keeping my lunch in the cryostat. The fresh
unfixed tissue adds a certain je ne sais quoi to the flavor. :/ Sent
from my Verizon Wireless BlackBerry

-Original Message-
From: Lynette Pavelich lpave...@hurleymc.com

Date: Wed, 22 Jul 2009 18:07:07
To: lei...@buffalo.edu; histonet@lists.utsouthwestern.edu;
cing...@uwhealth.org
Subject: RE: [Histonet] Start Up Lab


Gosh.I remember the days sipping on my coffee and nibbling on a
fresh donut as I cut my morning slides!  Sigh..

 Merced M Leiker lei...@buffalo.edu 07/22/09 5:00 PM 
(lol some labs have a bench area as well as a desk area where food is
allowed.)

--On Wednesday, July 22, 2009 3:19 PM -0500 Ingles Claire
cing...@uwhealth.org wrote:

 In the lab?!? For shame. :)

 

 From: histonet-boun...@lists.utsouthwestern.edu on behalf of Cindy
DuBois
 Sent: Wed 7/22/2009 10:29 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Start Up Lab



 And a coffee pot.

 

[Histonet] LR White and immunofluorescence

[Va Paula Sicurello]

Happy Friday Everybody,

I have a user who has a precious sample and wants to embed in LR White, maybe.  
I'm having him practice on something not precious first.  My questions are:

1.  He wants to do immunofluorescence and morphology on these nerve samples.  
He is planning on perfusing in 2% PFA, 0.1% glut in 0.1M sodium cacodylate.  
Then he will dissect out the nerve, cut it in half and place half in 2% PFA, 
0.1% glut (embed this in LR White) and the other half in 4% PFA, 2.5% glut 
(embed this in Epon) to finish the fixation.  Will that work? Or will the 
perfusion fixation with the weaker fix cause the stronger submersion fix to be 
moot?

2.  He wants to do immunofluorescence on the LR white sample.  I figure, 
primary antibody, then secondary with glow is no different that secondary with 
gold ball, am I correct in this?

Thanks and have a great weekend.

All aglow waiting for your answer,

Paula :-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center (CRIC)
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

Your images flow through our CRIC.


  

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Re: [Histonet] Frozen Artifacts

[Va Paula Sicurello]

Hi Nicole,

You must tell the list what the artifacts are before we can help you.

You're note is a little too vague.

Paula Sicurello

VA Medical Center San Diego

Veterans Medical Research Foundation (VMRF) 

Core Research Imaging Center (CRIC)

3350 La Jolla Village Dr., MC151

San Diego, CA 92161

858-552-8585 x2397



Your images flow through our CRIC.

--- On Fri, 6/5/09, Patten, Nicole (NIH/NIAAA) [F] patte...@mail.nih.gov 
wrote:

From: Patten, Nicole (NIH/NIAAA) [F] patte...@mail.nih.gov
Subject: [Histonet] Frozen Artifacts
To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu
Cc: Rodriguez-canales, Jaime (NIH/NCI) [F] rodri...@mail.nih.gov
Date: Friday, June 5, 2009, 7:46 PM

Hello Histonet...

I am using frozen brain sections and I have terrible frozen artifacts. I 
receive the tissue from a brain bank so I have no control over the method of 
freezing. Is there ANYTHING that I can do at this point to at least reduce 
these artifacts? I've tried formalin fixation post-cutting and reducing the 
temperature of the cryostat but nothing works the others in my lab also 
have no idea. Any help would be GREATLY appreciated. Thanks!!

-Nicole

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Re: [Histonet] Fixation for EM

[Va Paula Sicurello]

Hi Chana,

I'm going to highlight my answers after each of your questions below.

Though I will say that you should contact whomever may be processing the brains 
for EM to see what they would prefer.  Also ask them which buffer they prefer.  
Karnovsky's fix is traditionally made with Sodium cacodylate (an arsenic base 
compound) but some labs have switched to phosphate buffer for safety reasons.

If you have any other questions, please feel free to contact me.

Paula  :-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397

http://www.vmrf.org/researchcenters/confocal/confocal.html



 
 My questions follow:
 
 (1) What is the best fixative solution to be used under
 the
 circumstances? I was thinking of using a combination of
 paraformaldehyde and acrolein due to acrolein's penetrative
 qualities
 and superior fixative strength. 

 I think most labs tend to stay away from acrolein (it is tear gas and not 
pleasant to be around).  A modified Karnovsky's is probably the best all around 
general EM fixative.  If it's OK to mince the brain into 1mm cubed bits that 
would be best for proper fixation.  If you can't then you end up with a bit of 
a fixation barrier.  Glutaraldehyde penetrates at 0.5mm/hour at room temp.  
When the outside parts get fixed first and the sample is large, then fix for 
longer periods of time.  Samples can be left in the refrigerator for weeks.
 
 (2) Do I need to perform a secondary fix with osmium
 tetroxide
 myself...or do I leave that to the EM lab techs to perform
 when they
 make slices? If at all possible, I want to minimize the
 amount of
 tissue processing on my end.

Unless you have to, I would avoid the Osmium textroxide.  A good EM lab would 
prefer that you just fix your samples and either submit the samples in buffer 
or fixative.  While acrolein is tear gas, Osmium textroxide is seriuosly 
poisonous.  Exposure to it can cause blindness and death.  With an exposure 
limit of 0.002ppm it is nasty.  The one good thing is you will go blind before 
you stop breathing and the blindness only lasts a few months.  So if you can't 
see or things get blurry, it's time to get some fresh air   ;-).  
    Seriously, let the EM people do the Osmication.

 
 (3) How long can the brains be left in post-fixative
 solution prior to
 further processing in the EM lab? Or, in other words, is
 there a
 maximum time period after which fixed whole brains cannot
 be sliced,
 washed, embedded, or otherwise processed?

Theoretically, samples can be left in fix a long time.  I once forgot a 
monolayer of cells and left them in Karnovsky's for 2 weeks without any 
noticeable deterioration of the ultrastructure.  
Again, ask the EM people, some have protocols they like to follow and times 
they prefer for optimization of image quality.
 
 (4) Any other suggestions, comments, etc.? Obviously, there
 has to be
 a less-than-ideal division of labor in this matter because
 we do not
 expect the EM lab to remove brains from rats that have been
 dead for
 up to 48 hours. I can handle that part, but I want to
 ensure I do
 everything right because what I do affects the rest of this
 study.

Most EM labs prefer that the user do a minimum of processing to the samples 
submitted.  We are a superstitious bunch and like to be able to control almost 
the entire embedding process.  That way if something gets messed up we can 
figure out what we might have done wrong and correct it.
 
 Thank you for your time and any valuable insights into this
 matter.
 
 Chana de Wolf
 Advanced Neural Biosciences, Inc.
 




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Re: [Histonet] Fixation for EM

[Va Paula Sicurello]

Oops,  My highlighted portions didn't make it through the filter.  The answers 
are after each question.

:-}

Paula Sicurello

VA Medical Center San Diego

Veterans Medical Research Foundation (VMRF) 

Core Research Imaging Center

3350 La Jolla Village Dr., MC151

San Diego, CA 92161

858-552-8585 x2397






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[Histonet] OptiQuip Model 1200

[Va Paula Sicurello]

Hello Everyone,

In my continuing quest to find out how to finish putting the bulb assembly back 
together on my fluorescence microscope I've found out some additional 
information.

The power supply and bulb assembly were made by a company called OptiQuip.  I 
have called the company, which still exists, but have yet to receive a return 
phone call.

Anyway, if you have such a beasty and have put the bulb assembly back together, 
it holds either a xenon or mercury bulb, please let me know.  

Even OptiQuip's owner's manual does not have a drawing or explain it clearly 
enough for me to understand.

Once more with feeling,

Paula  :-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


  


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[Histonet] Biowave DFR-10 owner's manual

[Va Paula Sicurello]

Hello All,

I have inherited a Biowave DFR-10 (a fancy Ted Pella microwave) but I did not 
inherit the owner's manual.

Does anyone have a copy that they can send me?  

Thanks and have a nice weekend.

Paula  :-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


  


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Re: [Histonet] Large coverslips?

[Va Paula Sicurello]

Hi Yvan,

Does it have to be a coverslip?  There are some mounting media that harden and 
form a barrier with optical qualities similar to that of glass.

It might not be a perfect solution, but it might work.

I think the stuff I used was called Crystal Mount (?).  

Paula  :-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr.., MC151
San Diego, CA 92161
858-552-8585 x2397


--- On Tue, 4/7/09, yvan lindekens yvan_lindek...@yahoo.com wrote:

 From: yvan lindekens yvan_lindek...@yahoo.com
 Subject: [Histonet] Large coverslips?
 To: histonet@lists.utsouthwestern.edu
 Date: Tuesday, April 7, 2009, 9:05 AM
 
 Hi all,
 
 I’m looking for some kind of a DIY coverslip or a tape,
 plastic foil… anything usable to cover some large slides
 (+/- 4.5cm * 15cm = 1.8inch * 5.9inch) containing botanical
 and zoological sections mounted in Canada Balsam.
 
 Does anyone has a (cheap…) solution for this? It
 doesn’t need to be pristine optical quality as the slides
 are primarely intended to be used on an overhead projector
 in the class room, but the possibility of viewing them under
 low power (40x - 100x) would be a real advantage. 
 
 Thanks in advance four your wisdom and knowledge!
 
 Yvan.
 
 
 
 
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Re: [Histonet] Let's call a truce

[Va Paula Sicurello]

Boy Bernie,

See if I ever buy your albums again!

;-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


--- On Thu, 4/2/09, Bernie Taupin bernietau...@ymail.com wrote:

 From: Bernie Taupin bernietau...@ymail.com
 Subject: Re: [Histonet] Let's call a truce
 To: HistoNet histonet@lists.utsouthwestern.edu
 Date: Thursday, April 2, 2009, 4:04 AM
 At this point,  I think about 20
 or more different people have been involved in this thread!
 
 Would anyone else like to chime in with their two-cents
 about being on-topic or offer their inane peanut-gallery
 observations and silly tit-for-tat bickering like Paula
 here? 
 
 
 
 
 From: Va Paula Sicurello vapat...@yahoo.com
 To: HistoNet histo...@lists..utsouthwestern.edu
 Sent: Wednesday, April 1, 2009 1:10:30 PM
 Subject: [Histonet] Let's call a truce
 
 
 Hello Netters,
 
 Time to stop the current off topic conversations and get
 back to histology.
 
 I apologize to Bernie for my comment since I contributed to
 this off topic topic.
 
 It's a new month let's start fresh.
 
 Happy slicing and dicing and may all your stains work
 perfectly.
 
 Paula  :-)
 
 Paula Sicurello
 VA Medical Center San Diego
 Veterans Medical Research Foundation (VMRF) 
 Core Research Imaging Center
 3350 La Jolla Village Dr., MC151
 San Diego, CA 92161
 858-552-8585 x2397
 
 
       
 
 
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[Histonet] Let's call a truce

[Va Paula Sicurello]

Hello Netters,

Time to stop the current off topic conversations and get back to histology.

I apologize to Bernie for my comment since I contributed to this off topic 
topic.

It's a new month let's start fresh.

Happy slicing and dicing and may all your stains work perfectly.

Paula  :-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


  


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Re: [Histonet] onalighternote

[Va Paula Sicurello]

Hey, we can't help it if your parents were big fans of his.  He is a great song 
writer.

;-)


Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


--- On Tue, 3/31/09, Bernie Taupin bernietau...@ymail.com wrote:

 From: Bernie Taupin bernietau...@ymail.com
 Subject: Re: [Histonet] onalighternote
 To: Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov, Edwards, R.E. 
 r...@leicester.ac.uk, histonet@lists.utsouthwestern.edu
 Date: Tuesday, March 31, 2009, 4:52 PM
 Yeah, hilarious.. I swear, I haven't
 heart THAT one before 
 
 
 
 
 
 From: Bartlett, Jeanine (CDC/CCID/NCZVED) j...@cdc.gov
 To: Edwards, R.E. r...@leicester.ac.uk;
 histonet@lists.utsouthwestern.edu
 Sent: Tuesday, March 31, 2009 10:18:36 AM
 Subject: RE: [Histonet] onalighternote
 
 I wanted to ask how Elton was but thought that was a bit
 too easy...
 
 
 
 Jeanine Bartlett
 Infectious Diseases Pathology Branch
 (404) 639-3590 
 jeanine.bartl...@cdc.hhs.gov
 
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu]
 On Behalf Of Edwards,
 R.E.
 Sent: Tuesday, March 31, 2009 10:16 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] onalighternote
 
 
 I have  just  had  s from  Bernie
 Taupin and Paul Schofield, is
 this  a  record??.
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu]
 On Behalf Of Robyn
 Vazquez
 Sent: 31 March 2009 15:08
 To: Bernie Taupin; histonet@lists.utsouthwestern.edu
 Subject: RE: [Histonet] Ford Royer
 
 I think Ford gets the message...GET ON WITH IT! 
 Antibody talk anyone?
 
 -Original Message-
 From: histonet-boun...@lists.utsouthwestern.edu
 [mailto:histonet-boun...@lists.utsouthwestern.edu]
 On Behalf Of Bernie
 Taupin
 Sent: Tuesday, March 31, 2009 6:50 AM
 To: histonet@lists.utsouthwestern.edu
 Subject: [Histonet] Ford Royer
 
  I can personally attest that Ford must have been
 having a VERY bad day
 indeed.
 
 Although I can empathize that this might be the case, I do
 find it
 curious that we've heard nothing from Mr. Royer since his
 little
 outburst.
 
 I, for one, wouldn't mind an apology to the list for your
 antisocial
 behaviour on it, Mr. Royer.
 
 
 
       
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Re: [Histonet] incubator oven

[Va Paula Sicurello]

Hi Kris,

I think that a smallish lab oven would suffice.  If all you are doing is 
melting paraffins I think any of the ones from the major vendors (Fisher, VWR, 
etc.) would be good enough.

I do believe that those are still just ovens and don't have the air currents.

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


--- On Wed, 3/11/09, Kalleberg, Kristopher kristopher.kalleb...@unilever.com 
wrote:

 From: Kalleberg, Kristopher kristopher.kalleb...@unilever.com
 Subject: [Histonet] incubator oven
 To: histonet@lists.utsouthwestern.edu
 Date: Wednesday, March 11, 2009, 1:40 PM
 Hi All,
  
 I desperately need to get a new incubator oven for my
 histology lab.  It
 seems as if most ovens are now convection ovens. 
 Since my old oven is
 not convection I am just concerned that the constant air
 movement will
 some how affect the tissue slides in the way the paraffin
 in melted
 before the processing of the slides.  I will be using
 this oven for
 strictly melting paraffin containers to have ready for
 embedding center
 and for baking of slides to melt paraffin prior to
 processing.  Has
 anyone had any concerns or issues with these new convection
 ovens or can
 they recommend a decent oven to buy.  it just needs to
 be a medium sized
 oven.  
 Thanks in  advance.
  
 Kris kalleberg
 kristopher.kalleb...@unilever.com
   
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[Histonet] Bad sections

[Va Paula Sicurello]

Hi Histo netters,

I am am my wit's end.  I know how to section but my sections are compressing 
like crazy.  I've altered my processing protocol thinking that I was 
overprocessing my samples.

I've tried different knife angles, different brands of razor blade knives.

It's either me or the microtome.

Info- I process animal tissues with between 30-60 minute steps.  Vacuum 
paraffins for 30 minutes.  Section with Reichert-Jung 2030 set at 4-ish degrees 
(a little below the middle line on the knife holder).
Soak blocks on Downey ice blocks, water bath at 44 degrees.

Suggestions?  

Waa,

Paula

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


  


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[Histonet] Cutting angle?

[Va Paula Sicurello]

Hello Listers,

I have inherited a Reichert-Jung 2030 microtome.  The knife holder is a bit 
funky and hard to set the clearance angle.

What angle do y'all use?  I'm trying to get 4 or 5 degrees.

Thanks,

Paula :-)

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


  


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Re: [Histonet] problem eyes

[Va Paula Sicurello]

Really?  Liquid laundry soap?  Does this help with all wrinkles in large 
specimens?


Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Microscope Facility, room B141
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


--- On Wed, 3/4/09, Rene J Buesa rjbu...@yahoo.com wrote:

 From: Rene J Buesa rjbu...@yahoo.com
 Subject: Re: [Histonet] problem eyes
 To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu, 
 Perry, Margaret margaret.pe...@sdstate.edu
 Date: Wednesday, March 4, 2009, 9:14 PM
 Add a few drops (4-5) of liquid
 detergent, but not dishwasher detergent (because it will
 dissolve the paraffin).
 rené J.
 
 --- On Wed, 3/4/09, Perry, Margaret margaret.pe...@sdstate.edu
 wrote:
 
 From: Perry, Margaret margaret.pe...@sdstate.edu
 Subject: [Histonet] problem eyes
 To: histonet@lists.utsouthwestern.edu
 histonet@lists.utsouthwestern.edu
 Date: Wednesday, March 4, 2009, 4:04 PM
 
 Please help!  We have been trying to cut whole eyes
 from a pig.  They need to be
 nice enough for a publication.  We are having the
 devil of a time because no
 matter what we do there are wrinkles.  If we turn the
 waterbath up to stretch
 things out the retina detaches.  Do any of you have
 suggestions?  We uses
 Surgipath  EM400 for embedding paraffin and and their
 infiltration medium.  Is
 this to soft? The lowest we can turn our waterbath is 38
 degrees C.
 
 Margaret Perry HT (ASCP)
 IHC Lab Manager Veterinary Science
 Animal Disease Research and Diagnostic Lab
 South Dakota State University
 Box 2175 North Campus Drive
 Brookings SD 57007
 
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[Histonet] Sectioning Probems, again

[Va Paula Sicurello]

Hello Listers,

I have embedded mouse hearts into Surgipath EM-400.  The hearts are dropping 
out or rolling up and the sections are rolling onto themselves, not forming 
ribbons.  The hearts were given to me in 70% ethanol after sitting in it for an 
unknown time.

The sections are acting like it's dry outside yet it's rainy.  I don't 
currently have access to an ice block with Downey in it.

Other than soaking the blocks overnight in water (I'm trying that tonight) are 
there any other suggestions?

Thanks,

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Microscope Facility, room B141
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


  


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Re: [Histonet] Sectioning Probems, again

[Va Paula Sicurello]

For all those who wondered how I embedded my mouse hearts that won't section-

The processing schedule is as follows:

Tissue samples hold in 70% alcohol until the process begins using an 
Autotechnicon.  If you don't know what that is, ask someone who has been doing 
histology for a lng time.
80%, two 95's, 3 100's, 3 Citrisolve all for 1 hour with agitation.  2 changes 
of Surgipath EM-400 (same as I've used in clinical labs).  Then 30 minutes to 
one hour in EM-400 under vacuum.

Since I inherited all the reagents and embedding paraffins's and I have to get 
the histology core running with minimal expense (I know, I know, but tell that 
to the CEO) I must work with the reagents I have.

I can tel you that liver samples were embedded using the same reagents and wax 
and they sectioned fine.  Well, fine for liver.

Thanks for any input you can give.

Paula

Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Microscope Facility, room B141
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


--- On Wed, 2/18/09, Paula Pierce cont...@excaliburpathology.com wrote:

 From: Paula Pierce cont...@excaliburpathology.com
 Subject: Re: [Histonet] Sectioning Probems, again
 To: vapat...@yahoo.com
 Date: Wednesday, February 18, 2009, 12:19 AM

 Sounds like incomplete processing. What is your processing
 schedule?
 



  


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