Please read entire procedure before staining sections. Standard Immonuhistochemical Staining Procedure for Frozen Sections When ready to stain, remove slides from freezer and warm to -20☌ in the cryostat or -20☌ freezer, fix for 2 minutes in cold fixative (acetone or other suitable fixative) and allow to come to RT to continue with the staining.
Alternatively, the frozen section slides can be stored for a short period of time at -70☌ in a sealed slide box. alcohol, formal alcohol, formalin, etc.), air dry at RT and proceed to staining. 
Fix slides by immersion in cold acetone (-20☌) for 2 minutes or other suitable fixative (e.g. Cut sections of the desired thickness (usually 5 µm), place the sections on a Fisher Superfrost slide and dry overnight at room temperature (RT). Cut tissue block until the desired tissue is exposed. Adjust the positioning of the block to align the block with the knife blade. Place the tissue block on the cryostat specimen disk. Before cutting sections, allow the temperature of the block to equilibrate to the temperature of the cryostat (typically -20☌). Store blocks in the -80☌ freezer until ready for sectioning. NOTE: If block is left in 2-methylbutane too long, the block may crack. Allow the tissue matrix to solidify completely and remove block from 2-methylbutane and place on dry ice or in the -20☌ cryostat. Place base mold with tissue into the beaker of cold 2-methylbutane and quickly immerse the block. Place a stainless steel beaker of 2-methylbutane in liquid nitrogen and allow to cool adequately. Arrange tissue in the matrix near the bottom so tissue is easily exposed when sections are cut. Place in pre-labeled base molds filled with frozen tissue matrix. Remove desired tissues, trim and cut tissue no more than 5 mm thick. Sacrifice animal by prescribed and approved euthanasia techniques. Label base mold and partially fill the mold with frozen tissue matrix. Frozen tissue matrix (OCT® or Cryomatrix®). Preparation and Staining of Frozen Tissue Sections
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