[Histonet] Problems with sectioning hairy skin

[Shanu Mehta]

Hello All,

I have been sectioning some fresh frozen hairy human skin on the cryostat and I am having too much trouble getting decent sections. The sections seem to tear up and the epidermis & the dermis appear to separate.

My technique:
I embed dermis side down in OCT compound and then place the mold onto some dry ice until the mold solidifies. I then transfer the molds to a -80ºC freezer until they are ready to cut. Next I let the blocks come to cryostat temperature in the chamber. For cutting, I turn the block at a 90º angle meaning that both the dermis & the epidermis get cut in the same plane. I usually cut at a Chamber Temp of -19 & an Object Temp of -21. My cutting angle is between 3 & 5. I use low profile blades. I trim most of the fat that may be present in the skin, but there may be residual fat pockets that I may have missed!

This morning I talked to the technical staff at Leica and I was suggested to change the Chamber Temp to -21 to match that of the Object Temp. Also, I was asked to try with an angle of 5 or greater than 5 (as high as 8 or 9). I tried a range of temperatures and cutting angles, all in vain. I am still not happy with the quality of sections I am getting! 

Any suggestions on what I could do? I greatly appreciate any comments.

Thanks to all,
Shanu

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Shanu Mehta
Magen Biosciences
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Waltham, MA 02453
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Email: smehta <@t> magenbiosciences.com
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