The Histology and Immunohistochemistry Laboratory
Department of Pathology, Room 325C
The University of Texas Health Science Center at San Antonio
San Antonio, Texas 78229-7750
Phone: (210) 567-6599
Fax: (210) 567-6602
Label all containers with patients name and STRL Laboratory Request Form.
Biopsy must be received fresh, not denatured in any fixation agent. The biopsy may be received in the laboratory wrapped in a saline-dampened gauze pad to prevent drying, contractile artifacts and the loss of possible soluble enzyme activity or macromolecules like glycogen.
The biopsy should be kept chilled with ice cubes or chips in a dry closed container in the refrigerator at 40°F(4C). The biopsy must be quick-frozen within l-2 hours. If the biopsy is being transported to a core facility within the same city the closed container can be placed into a bucket of ice cubes or chips. Do not place the container in the freezer of the refrigerator or in a styrofoam container, containing dry ice.
If the muscle biopsy is being shipped via Federal Express or equivalent, freeze the muscle biopsy as indicated below and pack in a styrofoam container with an ample supply of dry ice. The electron microscopy (EM) specimen (in glutaraldehyde) must accompany the biopsy in a separate container. Do not freeze the EM sample in the dry ice container. Patient and insurance information must accompany the biopsy. Label the package Medical Specimen.
Freezing the muscle biopsy can be successfully achieved in l0-30 seconds. Using 50-75 ml of fresh chilled 2-methyl butane (isopentane) chilled to l30 degrees centrigrade (check with a low grade temperature thermometer) in a plastic beaker slowly lowered into a dewar flask filled with liquid nitrogen. Small white plaque forms on the bottom and sides of the beaker at approximately-l00 degrees C. Do not allow the liquid nitrogen to bubble into the isopentane. Carefully picking up the coated biopsy with forceps, lower it into the l30 degree isopentane, wearing freezer gloves to protect your hands. Depending on the muscle measurement the biopsy is flash frozen in l0-30 seconds. Remove the biopsy from the plastic freezer container and store at -70 degree with the proper patient identification until cryotomy is performed. The flash frozen biopsy must not be allowed to thaw. If a low temperature freezer is not available, the frozen block can be floated on a styroform boat in liquid nitrogen until proper cold storage can be found.
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