Laboratory Test Directory

MUSCLE BIOPSY INFORMATION SHEET

Specimen: Skeletal muscle biopsy (fresh, unfixed) for diagnosis of neuromuscular disease.

• Requirements: (More Details to follow)

DO:

• Page the neuropathologist on call as soon as possible after the procedure is scheduled, since advance preparations are required.
• Obtain a muscle biopsy clamp from the Surgical Pathology Gross Room. The clamp is for a small fascicle for electron microscopy that is separate from the larger piece for histology and histochemistry.  Both non-sterile metal clamps and single-use pre-sterilized plastic clamps are available.
• Provide two pieces of unfixed muscle - one single fascicle in the clamp (< 1 mm diameter) and a second larger unclamped piece (min. size 0.3 x 0.3 x 0.5 cm). This is the minimum amount of tissue for routine procedures.
• Wrap the tissue with a barely-moist gauze to prevent drying and cool the sealed specimen container with water/ice during transport.
• Page the neuropathologist on call when the biopsy is ready and deliver the tissue to the Surgical Pathology Gross Room within 15 min. after excision. Mark "STAT - FOR NEUROPATHOLOGY"

DO NOT:

• Do not immerse the tissue in formalin or saline!

• Do not leave the tissue in Surgical Pathology or in the Surgical Pathology refrigerator after hours. After hours you must page the neuropathologist on call before you obtain the specimen to arrange collection of the biopsy.

What diagnostic services are available?

1. Muscle histology (routine)
2. Muscle histochemistry (routine, selected by pathologist)
3. Immunohistochemistry and immunofluorescence (special studies)
4. Electron microscopy (special study)
5. Computer-assisted muscle fiber morphometry (special study)
6. Transfer of muscle tissue to outside laboratories for certain special studies

What do we do routinely?

1. Light microscopic diagnosis based on selected histologic and histochemical stains - H&E, oil red-O for neutral lipids, modified Gomori trichrome, PAS, NADH-TR (NADH-tetrazolium reductase) and ATP-ase at pH 9.6, 4.6 and 4.35.
2. We routinely fix the thin fascicle in the muscle clamp in gluteraldehyde/paraformaldehyde fixative and hold it in case electron microscopy (EM) is necessary. EM is a separate study (at considerable additional cost).
3. We routinely freeze and hold at -80°C any additional tissue not used for the above studies in case special biochemical or molecular biologic studies by outside laboratories are needed (e.g. analysis of dystrophin, mitochondrial enzymes or mitochondrial DNA). We can arrange to ship frozen muscle tissue to outside laboratories, but the submitting physician is responsible for supplying sufficient tissue and completing the required clinical data and billing forms.

Special studies in addition to the routine procedures described above are done at the discretion of the neuropathologist and carry additional charges. If special studies are specifically requested, please provide adequate clinical data to justify the performance of these studies.

Information Required on the Tissue Consultation Request:

1. Patient’s name, UH #, ICD9 code and financial account number.
2. Date and time of excision of biopsy.
3. Attending physician’s name, doctor #, page and telephone numbers and address to send report.
4. Names and addresses of any other physicians to receive reports. Please include the names of the neurology and/or neurosurgery consultants.
5. Clinical diagnosis (both neuromuscular disease and any other relevant diagnoses).
6. Clinical history, physical findings and laboratory data relevant to neuromuscular disease, including electromyographic (EMG) diagnosis and creatine kinase (CK) levels.

Collection and Processing Details:

1. Obtaining a muscle biopsy clamp: Many services do not keep muscle clamps in stock. If you do not have a clamp, please call the Surgical Pathology Gross Room to arrange for loan of an appropriate clamp. You can then send a messenger to pick up the clamp to the Surgical Pathology Gross Room prior to the procedure. The reusable metal clamps are not supplied sterile and must be autoclaved before use. If you send your own clamp, your messenger can pick it up on the day following delivery of the biopsy.
2. Requirement for rapid transport to the laboratory: The unclamped portion of the biopsy must be frozen in the laboratory under special conditions and the clamped piece must be placed in EM fixative as soon as possible after removal from the patient, generally within 30 minutes, but the shorter the delay the better. These requirements are necessary to preserve the enzymatic activity of the muscle tissue, which is required for successful histochemical staining, and for optimal preservation of light microscopic and ultrastructural (EM) morphology.
3. Delivery to the laboratory: Most muscle biopsies are performed in the O.R., and in most respects muscle biopsies should be treated in the same manner as frozen sections. Muscle biopsies should be delivered to the Surgical Pathology Gross Room, during the hours 8 a.m. - 5 p.m., M-F. Please mark the biopsy "STAT" and notify the gross room technician that the biopsy is for Neuropathology. At any other time you must page the neuropathologist on call to arrange for collection of the biopsy. If you are performing a muscle biopsy outside of the O.R. suite and/or the procedure is not on the O.R. schedule, it is imperative that you call the Neuropathology office or page the neuropathologist on call prior to the procedure, since processing this tissue requires certain advance preparations.
4. Turn-around Time: Standard time for processing, staining and interpreting the slides is 7 days, but time may vary depending on the week day received and any additional studies that may be required for diagnosis Please call the neuropathologist for information on the status of the report. A preliminary verbal report covering only the presence or absence of inflammation for cases of suspected inflammatory myopathy can generally be made by the next day if specifically requested.

 FOR FURTHER INFORMATION PLEASE CALL: Gerald A. Campbell, Ph.D., M.D., Division of Neuropathology, UTMB at the above telephone number.

Revision Date: 10/14/04 Reviewed 06/22/12
Replaces Rev. of: 3/21/02