| Place the cores in a small amount of saline in a Petri dish or on saline-moistened gauze so it won't dry out while you divide the tissue. Handle the tissue only on a CLEAN surface (a wax plate is appropriate). Use a new, clean razor blade to divide the tissue (a double edge razor is appropriate). Do not pick up the tissue with forceps - it will squeeze and distort the tissue. Instead, roll the pieces up on a thin wooden stick (the end of a cotton tip applicator works fine). The tissue will adhere to the wood, and then insert it into the vial with fixative and gently swirl/shake the tissue off of the stick. Use a new stick for each fixative so that the fresh tissue is not contaminated with fixative carried over from the vials. Check that the tissue does not stick to the side of the container or under the lid.
To optimize dividing the tissue, visualize it under a dissecting scope or with a hand held magnifying glass, you can see tiny little red dots-these are the glomeruli. Cut off two consecutive 1 mm pieces from each end of each core for EM and IF, respectively. (Follow the protocol below to optimize allocation of tissue.) Put the remaining pieces of tissue in fixative (10% formalin) for LM.
1. Three cores are recommended for diagnosis and morphologic studies. Divide 2 cores according to the protocol below. Save the third core entirely for LM.
2. Place the tissue for light microscopy in 10% formalin. Place the tissue for immunofluorescence in Zeus fixative. Place the tissue for electron microscopy in 3% glutaraldehyde.
3. A diagram is available.
For consultation on slides prepared by another laboratory: stained slides appropriately identified with the name of the laboratory where prepared and accession number along with immunofluorescence results and electron microphotographs if available.
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