Efficacy of Transplant Media for Muscle Biopsy Sample Preservation
Kunle O Ojemakinde, Jon D Wilson. Louisiana State University Health Sciences Center, Shreveport, LA
Background: One of the limitations in the pathologic evaluation of muscle biopsy is the time interval between the biopsy procedure and the snap freezing of the sample. Timely freezing of the sample in liquid nitrogen cooled isobutane not only gives excellent frozen section morphology, but also preserves tissue biochemistry and nucleic acids. The latter allow for accurate enzyme histochemical staining and/or subsequent biochemical and genetic testing. Institutions that refer muscle samples to a specialist are often too far to transport these time-sensitive specimens by courier and/or do not have the capability to snap freeze the muscle biopsy on premises.
Transport media are used to preserve organ donor explants during shipment. A prior report indicates that use of transport medium preserved the enzyme histochemistry of pig muscle biopsy samples for up to five days. We assessed the preservation quality of two transport media on human muscle.
Design: Skeletal muscle tissue was sampled from freshly amputated leg specimens. The samples were divided into five portions in each of the two transport mediums (“Belzer UW Cold Storage Solution” and “Lifor ACF perfusion media”), and Carnoy's fixative. One portion was snap frozen in liquid nitrogen cooled isobutane immediately. The remaining four were placed in a sterile conical tube containing 50ml of transport medium or Carnoy's fixative, and refrigerated at 4 degrees centigrade. One portion of muscle per each successive day (days 2 through 5) was snap frozen from each transport medium or preservative. Frozen sections from each of the five samples were cut and stained at the same time. Staining included hematoxylin and eosin, modified Gomori Trichrome, PAS and PASd, and enzyme histochemistry. The enzyme histochemical stains included ATPase at pHs 9.4 and 4.3, NADH-TR, alkaline phosphatase, acid phosphatase, cytochrome oxidase, succinic acid dehydrogenase, and myophosphorylase.
Results: Both transport media preserved enzymatic activities and provided good frozen section morphology for up to 5 days. Compared to the standard frozen biopsy specimens, biopsies processed from transport media contained some artifacts, such as cell shrinkage and/or tissue edema. Lifor ACF perfusion media showed somewhat more tissue edema than Belzer UW Cold Storage Solution. The sample preserved in Carnoy's fixative did not perform well.
Conclusions: Our results confirm that organ transplant media represent a viable solution for preserving muscle biopsies for morphologic and enzyme histochemical assessment.
Tuesday, March 20, 2012 1:00 PM
Poster Session IV # 281, Tuesday Afternoon