[Histonet] looking for method for fixing frozen tissue

[Judi Ford via Histonet]

Hi everyone,
I was wondering if anyone can tell me the best way to formalin fix frozen 
tissue for processing into a paraffin block. I have tissue in a minus 80 
freezer and would like to cut a piece from it to fix in formalin and process. 
I'm thinking that I could put the tissue into the cryostat and bring it slowly 
up to minus 20. The container of formalin could be cooled down so that both are 
near the same temperature when I submerge the tissue into the formalin.  Are 
there other methods that may help process the tissue better?
Huge thanks to any suggestions that come my way.
Hope you all have a really good weekend.
Regards,
Judi




Judi Gordon
Senior Research Associate
Translational Sciences/IHC
CytomX Therapeutics


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[Histonet] Translational science/medicine/research IHC lab question

[Judi Ford via Histonet]

Hi Everyone,
I am interested in connecting with techs/managers who are working in or have 
developed a translation science/medicine/research IHC lab. Recently my 
department has become a translational science department and I run most of the 
IHC. I'm interested in building this lab into a good translational ihc lab and 
was wondering about the following:

1.   What does being a translational ihc lab mean?

2.   What advice would you have in building a translational ihc lab?

3.   What equipment would you want in a translational ihc lab that isn't 
necessarily in a regular research ihc lab?

4.   Are you available offline for further questions?
I really appreciate any guidance you send my way. I'm looking at this as a long 
term goal with the possibility of becoming the manager of the lab.
Thanks so much in advance for any advice.  Hope you all have a really great 
weekend :)
Best regards,
Judi Gordon
SRA
Translational Sciences
CytomX Therapeutics
South San Francisco, CA


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[Histonet] Tissue banking

[Judi Ford via Histonet]

Hi everyone, Happy Friday to you all..

I am hoping to gather some information on how people bank frozen tissue for 
work in research. I am trying to start a bank of frozen human samples stored in 
a -80 freezer for our scientists use.  Information that I'm looking for is as 
follows:

1.   Accessioning software/database - what software do you use to keep 
track of samples and their useage. What information is useful for you in 
keeping track of the samples.

2.   Storing samples in the freezer - how do you store them; wrapped foil, 
in cryoboxes, in ziplock baggies, etc.

3.   Do you have a system of checking out the samples? If so, how do you 
keep track and control of the samples?

4.   Do you have one person who is in charge of the tissue bank? If so what 
are their responsibilities?

Thanks for your help with this.  If you want to send me information offline 
here is my email address: jf...@cytomx.com.

Thank you and I hope you have a really great weekend.
Judi Ford

SRA
CytomX Therapeutics

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[Histonet] Sample accessioning software

[Judi Ford via Histonet]

Hi Everyone,
Happy Tuesday :).
My group is looking into software which would assign accession numbers, track 
histology specimens, list archive location, track disposition info, hold 
images, ability to attach scientific data, has security and to create reports. 
I would love to hear what anyone may be using or can suggest that I look into.
Thanks so much for any and all responses that come my way. I really appreciate 
it.
Best to all,
Judi Ford
CytomX Therapeutics,
South San Francisco, CA


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[Histonet] Freezing biopsies

[Judi Ford via Histonet]

Hi everyone,
Does anyone have experience working with human frozen biopsies? I have a 
question concerning freezing the biopsies. I am just doing general background 
research on this subject at the moment and have not worked with frozen human 
biopsies.

If a biopsy was taken using a 14 gauge needle, in general, what is the length 
and diameter of a biopsy and what would the preferred method of freezing be 
(flash freezing in dry ice or freezing in liquid nitrogen)? What is the method 
for flash freezing in dry ice? What gauge of needles are most commonly used?  
Are most biopsies taken using a 16-18 gauge needle?

Thanks for your help with this. I really appreciate it.
Best regards,
Judi Ford


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[Histonet] Special stain art work

[Judi Ford via Histonet]

Hi everyone,
I am interested if anyone has used histology special stain/IHC/IF images as 
artwork for hallways.  How large did you print the images, what resolution were 
they scanned at, etc.  Years ago I came across an article about someone who 
created hall panels of special stain artwork.  They created smaller image files 
at high resolution then stitched them together to create large (6'x6') panels 
to use in the hallways. Anyone remember seeing that article? Anyway, I would 
love hearing how people go about creating artwork with our images.
Thanks,
Judi
CytomX Therapeutics
South San Francisco, CA

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recipient, you have received this message in error and that any use, 
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copies of this message and any attachments.

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[Histonet] H staining on tape

[Judi Ford via Histonet]

Hi everyone,
I have a colleague that has been asked to do H staining on a frozen specimen 
collected on tape. The tape has been attached to the slide using nail polish. 
Can't use the CryoJane system due to the light flash. Her question is "will the 
nail polish disintegrate during the immersion in alcohol and xylene?". Another 
question is will the stains attach to the tape and obscure the tissue?  If so, 
what to do with about that. She is also worried that the tape will fall off 
into the staining dish and get lost. I'm thinking maybe using a 6 well 
disposable tissue culture plate for the staining of the tape then reattach it 
to the slide and coverslip.
Any suggestions?
Thanks so much,
Judi
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[Histonet] Staining stomach cells

[Judi Ford via Histonet]

Hi everyone,
I'm trying to differentiate parietal vs chief vs endocrine cells in the 
stomach. Any ideas on stains or antibodies I could use.
Thanks,
judi
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[Histonet] Image analysis software

[Judi Ford via Histonet]

Hi everyone,
We are in the middle of looking at image analysis software and I was wondering 
what software programs people are using. Why did you pick the one you are 
using? Are you using it for brightfield whole slide analysis or for fluorescent 
slide analysis?  What file types can be used with your software?

I would love to hear your experiences.

Thanks,
Judi
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[Histonet] Davidson's fixative and IHC question

[Judi Ford via Histonet]

Hi Everyone. Hope you all had a really good weekend. Thanks for the replies to 
my double ihc question.

I do have another question; this one is from a friend of mine. Her client wants 
to do ihc on rabbit eye tissue. The tissue will be fixed in Davidson's fixative 
for 24 hours then in 10% NBF. Will this have affect future ihc projects?

Thanks,
Judi
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[Histonet] Double stain IHC question

[Judi Ford via Histonet]

Hi everyone,
I have a question in chromogenic double staining. Here is the situation.
Tissue = human, frozen
Antibody = same protein (A)

1.   Commercial antibody of A

2.   Homegrown antibody of A, human, biotinylated

Question: can you stain both versions of this antibody on the same tissue, same 
slide? Goal is to see where each stains in the tissue and if they co-localize. 
If they do co-localize then how do you distinguish between that and where they 
stain individually? Would you use different chromogens and hope that where they 
come together it turns a different color?
I am really interested if this can work. Thanks in advance for any replies.
Judi
South San Francisco, CA
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