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Protocol for Sending Nerve Biopsy Specimens

  1. Please notify the Neuromuscular Pathology Laboratory two days in advance that a nerve specimen will be arriving. Please contact:
  2. A surgeon familiar with a diagnostic nerve biopsy procedure should obtain the specimen. Full thickness nerve approximately 4 cm in length is needed.
  3. The nerve is handled as follows. The nerve is placed on a 3x5 index card by the surgeon or the technician. DO NOT STRETCH THE NERVE. Work on the index card. Cut the nerve into 3 sections as detailed below with a double-edged super stainless blade on the index card.

    Section #1
    • Cut a 1 cm piece for section #1 and place it on the top left hand corner of the card.
    • Cut the card around the nerve and place it face down in a container of 4% paraformaldehyde prepared as follows: For 24 ml total solution (or 2 bottles), pour 12 ml 8% Paraformaldehyde and 12 ml 0.2M Sorensen's Phosphate buffer into a graduated cylinder.
    • Mix well by placing parafilm over top of cylinder and inverting several times.
    • Pour 13 ml of this solution into a biopsy fixative bottle and label as 4% Paraformaldehyde.
    • Use the remaining 11 ml solution to prepare the 3% glutaraldehyde/4% paraformaldehyde needed below.

    Section #2
    • Cut a 2 cm piece for section #2 and place it at the bottom of left hand corner of the card.
    • Cut #2 into 3 smaller pieces.
    • Cut the card around the nerve and place it face down in container marked 3% glutaraldehyde/4% paraformaldehyde prepared as follows: Into the remaining 11 ml of 4% paraformaldehyde solution add .7 ml of 50% Glutaraldehyde.
    • Mix well by stirring with a pipet. Pour this solution into a 2nd fixative bottle and label 3%Glutaraldehyde/4%Paraformaldehyde.

    Section #3
    • Pin one end of the remaining nerve section #3 (1-2 cm) to the index card and freeze it in 2-methylbutane (isopentane) in a metal cup surrounded by dry ice in a rubber bucket.
    • Cool the methylbutane in advance by dropping small pieces of dry ice into the methylbutane (see muscle protocol).
    • When the nerve segment is thoroughly frozen, pack section #3 in sufficient dry ice and the other sections in regular ice and ship overnight to:

      Christopher Dorsey or Jamal Garrison
      The Neuromuscular Pathology Laboratory
      Johns Hopkins Hospital
      600 N. Wolfe Street
      Meyer Building Room 5-119
      Baltimore, MD 21287-7519
      Phone: 410-614-4163
  4. The following information will be needed. Tissue samples cannot be processed unless this information is provided with the specimen.
    • Patient's name, address, date of birth, and hospital ID number
    • Hospital charge number for billing directly to the hospital
    • Brief clinical history
    • Identification of muscle biopsy site, date, and time
    • Name, address, phone and fax # of referring pathologist and attending physicians

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