Frozen Section Protocol

2 gently lift up while crossing the blade as the brush and block meet the blade the section will begin to form under the brush.

Frozen section protocol. Immunohistochemistry protocol for cryopreservation of tissues prior to fixation immediately snap freeze fresh tissue in isopentane mixed with dry ice and keep at 70 c. The frozen blocks can be temporary stored in dry ice. The technical name for this procedure is cryosection.

Place the tissue in a small sealed box if the tissue is too small to hold it pour in some embedding reagent like oct. Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining. Use frozen section to provide rapid gross or microscopic diagnosis to identify an unknown pathologic process identify extent of disease evaluate margins identify metastases or simply identify a tissue process tissue to provide appropriate and accurate diagnosis prognosis and to adhere to research and special study protocols.

1 ride the block as the block descends toward the brush the brush keeps pace with the block by gently resting on the bottom 2 3 mm of the block. From this point over the sample should never be thaw unless there is specific requirement. Paraffin and frozen sections reagents can be applied manually by pipette or this protocol can be adapted for automated and semi automated systems if these are available.

Cut cryostat sections. Dry the tissue sections overnight at room temperature. Transfer the blocks to a liquid nitrogen storage tank years or 80 c freezer months.

In my experience this practice adds to potential artifacts at the beginning of the section potential variations in thickness and leads to difficulties when approaching tissues containing fat. The quality of the slides produced by frozen section is of lower quality than formalin fixed paraffin embedded tissue processing. It is used most often in oncological surgery.

Sections can be stored in a sealed slide box at 80 c for later use. With the regressive method overstaining the tissue section with a neutral hematoxylin solution is the initial step. Frozen section protocol 1.