Labware

Protocol to make frozen section or paraffin-embedded section with cell-sheet.

When the section is made with the membrane attached to cell-sheet, the morphology of the cell-shet can be maintained as it is whereas the size of the section may become smaller due to shrinkage of the intranuclear frame when the section is made without the membrane.

A) Protocol to make frozen section

1. Following the protocol of cell sheet collection (→Standard Operating Protocol (SOP) for collection of cell-sheet.), please recover the cell-sheet.（*1）
2. Using knife, cut the cell-sheet to desired size.
3. Fill 50% of the cassette with OCT compound.
4. Using forceps, place the cell-sheet in the cassette.
5. Remove the membrane from cell-sheet, if required.
6. Fill remaining 50% with OCT compound.
7. Cool and fix the section.

*1 If required, fix the cell-sheet overnight at 4℃ containing 10% of formalin.

B) Protocol to make paraffin-embedded section

1. Following the protocol of cell sheet collection (→Standard Operating Protocol (SOP) for collection of cell-sheet.), please recover the cell-sheet.
2. Fix the cell-sheet overnight at 4℃ containing 10% of formalin.
3. Using knife, cut the cell-sheet to desired size.
4. Place cell-sheet in the cassette.
5. Wash off formaline from cell-sheet with iced water.
6. Place cell-sheet in the embedding machine. Place weight on the cassette.
7. Make paraffin block using cell-sheet penetrated with paraffin. (*2)

*2 Machine setting should be in line with the actual weight of cell-sheet.