Author Topic: Epidermal and Dermal folding in frozen skin sections  (Read 2240 times)

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Offline NealHall

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Epidermal and Dermal folding in frozen skin sections
« on: March 09, 2015, 10:12:07 AM »
I'm using a frozen horizontal sliding microtome to produce 50 micron sections of skin. After the antibody staining the sections are mounted from a PBS/10% triton solution using hypodermic needles onto glass positively charged and subbed slides. After counterstaining and manual coverslipping the sections continually have folds either in the stratum corneum, epidermis, dermis and or all three when viewed under the scope at 40X. Any tips hints or pointers to reduce or eliminate the folds?

Epidermal and Dermal folding in frozen skin sections
« on: March 09, 2015, 10:12:07 AM »

Offline gula

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Re: Epidermal and Dermal folding in frozen skin sections
« Reply #1 on: March 09, 2015, 10:49:26 AM »
Do you think the folds are from mounting or from cutting?
I have no experience with this special procedure, but I work regularly with sliding microtomes to cut paraffin slides.
To get a section with hopefully no folds, we orientate the dermal surface rectangular or with a declination about 120 to the knife's edge. The knife must be very sharp to avoid uneven pressure on the edge.

Times ago we used to put some small metal-weights on the coverslipped slides. This prevents the section of "curling up" while the mounting media is drying.
hope this helps a little bit.
gula

Offline NealHall

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Re: Epidermal and Dermal folding in frozen skin sections
« Reply #2 on: March 09, 2015, 11:02:51 AM »
Not really sure. I'm running on the assumption that its a problem with mounting since we typically will see tissue from the same biopsy that some will have folds while others won't. So unless theres variation in each pass of the blade I'm working from the side of the mounting with the hypodermic needles and the PBS/triton solution. We ran controls with and without pressure weights and found no difference in the folds using the weights. The one thing so far that has somewhat helped is after mounting and allowing the sections to dry for 10 minutes, we have been rinsing downwards across the epidermis using a PBS solution.

Offline WWang1991

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Re: Epidermal and Dermal folding in frozen skin sections
« Reply #3 on: June 01, 2015, 05:25:18 AM »
well, not very clear about how to help you directly solve this problem, but you may find help from them http://www.creative-bioarray.com/Products/Epidermal-Cells-list-137.htm.
The secret to happiness is to keep setting yourself new challenges.
http://blog.creative-bioarray.com/

Re: Epidermal and Dermal folding in frozen skin sections
« Reply #3 on: June 01, 2015, 05:25:18 AM »