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Author Topic: Fresh vs Old tissue sections  (Read 7660 times)

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Offline ASluo

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Fresh vs Old tissue sections
« on: April 15, 2008, 01:55:14 AM »
Hi guys,
Does anyone know if the freshness of the tissues impact dramatically on the morphology and staining intensity of the slides? I snap froze mouse colon embedded in OCT and got them fixed and sectioned (by our histology department) with a couple of days. The morphology and staining was really excellent considering them to be frozen sections! When I compared them with colitis tissues, they were absolutely a nightmare! I didn't prepare the colitis tissues but I was told the person snap froze them without cleaning them in TBS and did not remove the feacal material. They have been in the -80 freezer for about 6-7 months and I wondering if the time and the feacal material could have contributed to the poor morphology and the high background?!
Many many thanks,
Annie

Fresh vs Old tissue sections
« on: April 15, 2008, 01:55:14 AM »

Offline excalibur

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Re: Fresh vs Old tissue sections
« Reply #1 on: April 15, 2008, 05:47:05 PM »
The length of time being frozen is probably not a factor.

The fecal material probably is the culprit. Mouse chow will contain plant fragments which will play havok on the microtome blade and cause scratches and gouges.

They may need to be thawed and cleaned.

I am confused by your statement  "I snap froze mouse colon embedded in OCT and got them fixed and sectioned (by our histology department) with a couple of days." Did you mean they sectioned and then fixed the frozen sections on the slides?
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

Offline ASluo

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Re: Fresh vs Old tissue sections
« Reply #2 on: April 15, 2008, 08:23:50 PM »
Sorry about the confusion! I meant to say the tissues were snap frozen, sectioned and then fixed with acetone.
My boss think because they were frozen, sectioned and fixed within couple (about 3) days it made hell of a difference in terms of the morphology and staining. What are you thoughts?

Offline ASluo

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Re: Fresh vs Old tissue sections
« Reply #3 on: April 17, 2008, 05:39:37 AM »
I discussed the problem with the people who did sectioning and they say is unlikely the feacal material would've contributed to the poor morphology. They think the money is on repeated freezing and thawing the slides - freezing artefacts! What are your thoughts? thanks annie

Offline CanuckPhD

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Re: Fresh vs Old tissue sections
« Reply #4 on: April 17, 2008, 07:34:37 AM »
It would make it easier for us to offer suggestions if you posted some pictures of your colitis tissue. Then we can begin to see if it is freezing artifact, bad cutting etc..

Offline excalibur

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Re: Fresh vs Old tissue sections
« Reply #5 on: April 17, 2008, 08:31:30 AM »
I agree with CanuckPhD. Post a picture so we can see the tissue.

If the person freezing the tissue months ago did not bother to properly prepare the tissue, the freezing technique could have also been lacking.

I can say I have had to section tissue not properly cleaned of fecal material and scratches are a problem. Mouse chow is not knife friendly.  ;)
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

Offline ASluo

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Re: Fresh vs Old tissue sections
« Reply #6 on: April 17, 2008, 09:38:08 PM »
Thanks guys! but how do i post up images?  :D

Offline ASluo

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Re: Fresh vs Old tissue sections
« Reply #7 on: April 17, 2008, 11:20:53 PM »
Unfortunately the files are too large and can't be attached! >:(

Offline ASluo

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Re: Fresh vs Old tissue sections
« Reply #8 on: April 18, 2008, 07:21:55 AM »
I can send them to one of yous email address if yous want to have a look at them?

Offline excalibur

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Re: Fresh vs Old tissue sections
« Reply #9 on: April 18, 2008, 11:30:52 AM »
You may send them to me.
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

Offline richard03

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Re: Fresh vs Old tissue sections
« Reply #10 on: April 18, 2008, 06:15:39 PM »
You can register in Gallery (http://www.ihcworld.com/imagegallery/) and PM me your user name so I can activate your account and then you will be able to upload images after login.

Richard

I can send them to one of yous email address if yous want to have a look at them?

Offline ASluo

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Re: Fresh vs Old tissue sections
« Reply #11 on: April 19, 2008, 09:09:45 AM »
I've registered and username is ASLuo. many thanks! Annie :D

Offline richard03

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Re: Fresh vs Old tissue sections
« Reply #12 on: April 19, 2008, 09:31:10 AM »
Hi Annie,

Your account in Gallery is active now so please go ahead login and click upload then follow instruction to post your images.

I've registered and username is ASLuo. many thanks! Annie :D

Offline ASluo

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Re: Fresh vs Old tissue sections
« Reply #13 on: April 19, 2008, 10:28:18 AM »
Hey Richard,

I'm having problem with the posting. It won't let me post the images up....

Offline richard03

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Re: Fresh vs Old tissue sections
« Reply #14 on: April 19, 2008, 12:34:54 PM »
At which step have you stopped? The image needs certain format (.JPG is good, TIF is not). The max size is 100KB and no more. Let me know if you still have problem and post the details.

Hey Richard,

I'm having problem with the posting. It won't let me post the images up....
« Last Edit: April 20, 2008, 09:33:20 AM by richard03 »

Re: Fresh vs Old tissue sections
« Reply #14 on: April 19, 2008, 12:34:54 PM »