Author Topic: Trouble sectioning mouse pancreatic tissue  (Read 1255 times)

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Offline VShemica

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Trouble sectioning mouse pancreatic tissue
« on: January 06, 2015, 05:09:52 PM »
Hello everyone,

I looking for some help again, this time concerning the sectioning of formalin fixed paraffin embedded mouse pancreas.

The trouble is that during sectioning (7 microns-requested by researcher) the tissue appears dehydrated.  The tissue is "flackly" (used due to a lack of better words) for majority of each section.  Also, numerous small cracks/lines can be seen on the edges of the tissue when viewed under the microscope.

Before given to me, the tissue was prepped for IHCs by being fixed in 10% NBF for 24hrs, then stored in 70% ethanol for a "few days" (per researcher). ****I beginning to feel that extended periods of ethanol storage severely dries out the tissue, which is leading to troublesome sectioning****

Next I processed in a closed processor:
1)   85% Ethanol.......... 0:20 (No Heat)
2)   95% Ethanol.......... 0:20 (No Heat)
3)   100% Ethanol........ 0:20 (No Heat)
4)   100% Ethanol........ 0:20 (No Heat)
5)   100% Ethanol........ 0:20 (No Heat)
6)   Xylene.................... 0:20 (40oC)
7)   Xylene.................... 0:20 (40oC)
8)   Paraffin................... 0:30 (58oC)
9)   Paraffin................... 0:30 (58oC)
10) Paraffin................... 0:30 (58oC)
11) Paraffin................... 0:30 (58oC)

I have tried facing the tissue then placing on an ice bath for 5- 10 minutes, making sure the tissue doesn't turn white and swell.

Well everybody, all suggestions and comments will be greatly appreciated.  Hope to hear from you guys soon.

Thanks
« Last Edit: January 07, 2015, 12:13:42 PM by VShemica »

Trouble sectioning mouse pancreatic tissue
« on: January 06, 2015, 05:09:52 PM »