MUSCLE BIOPSY Information For Clinicians Information For Pathology Residents Muscle Biopsy Video Tutorial

Transcription

1 Modified by Dr. McKeever and Dr. Camelo-Piragua MUSCLE BIOPSY Information For Clinicians Information For Pathology Residents Muscle Biopsy Video Tutorial Procedure (for Main OR, Mott OR, Treatment Rm 3, and VAH): INFORMATION FOR CLINICIANS -Call the IPOX lab, Sylvia Zelenka, Lynn Tague or Susan Ahrens ( ), at least one day in advance if you plan to do a muscle biopsy, for the supplies and instructions. -If you have a question regarding appropriate muscle to biopsy contact neuropathologist on service: Dr. McKeever ( ; pager: 1426) or Dr. Camelo-Piragua ( ; pager: 17599). -If procedure is done after hours, contact the pathology resident on call ( ) before starting the procedure to assure the specimen is processed as soon as it is received. -It is the surgeon s responsibility to get the tissue to Pathology Department. A. OPEN SCALPEL BIOPSIES: Biopsies from adult patients (> 16 years of age) should contain 3 specimens: 1. One piece for histochemical stains, it should measure about 2 cubic centimeters. It should be gently removed from the belly of the muscle (avoid tendinous insertion!) and placed immediately in saline soaked gauze in a closed container. It should not be allowed to dry out or float in water or saline. It also should not be infused with excess lidocaine. 2. One piece for formalin fixation and paraffin embedding. It must be stretched on a tongue blade before immersion in formalin. It should measure about 2cm in length to allow stretching and tightening and at least 1.5cm in thickness. Please take care that there is enough formalin within the container. 3. One small piece for electron microscopy, similar in thickness to a match or a rounded toothpick. Attached to the sterile metal clamp in situ, this piece should not be teased or allowed to dry during its separation from the rest of the muscle. It should be immediately, within a few seconds, placed in a specimen jar full of glutaraldehyde if available. If not available in the OR, specimens (1 to 3) should be sent to pathology immediately, to process in a timely fashion. Biopsies from children (<16 years old) should have 2 separate pieces frozen as described in #1 above, and 1 small specimen for electron microscopy as described in #3 above. The formalin fixed portion (as in #2) is not required (however it is useful in cases of suspected inflammatory myopathy). B. NEEDLE MUSCLE BIOPSY: The entire biopsy is placed immediately on saline soaked gauze within a closed container and delivered to the Pathology Department where it will be frozen, except for a tiny bit that will be dropped in glutaraldehyde. No formalin fixed tissue is required. 1

2 INFORMATION FOR PATHOLOGY RESIDENTS Muscle Biopsy Processing for Histochemical, Ultrastructural and Metabolic/Genetic Analysis Review these resources in full prior to starting. If any question contact the muscle lab or the neuropathologist on service Resources The full procedure is posted in the muscle lab. You can find a full version of the on the intranet: In addition, a detailed video is available for review prior processing the specimen in the following link (courtesy of Dr. Limin Yu): Muscle biopsy video tutorial You may receive 3 containers/pieces scalpel biopsies > 16 years old ( See above information for clinicians): - #1 For histochemical analysis - #2 Formalin fixation and paraffin embedding. Place the tissue in a cassette and submit for regular histology. - #3 For electron microscopy: Put in Glutaraldehyde and place in the refrigerator (Room 2 behind Mary Joe s accessioning area) for the EM lab to pick up later. You may receive 3 containers/pieces scalpel biopsies, PEDIATRIC < 16 years old/ or Labeled: METABOLIC/GENETIC/FUTURE SEND OUT TEST - 1 piece should be frozen following Freezing procedure described below or in the links provided in Resources. - 1 piece can be placed in a cassette/cryotube (Label container with identifiers) and directly frozen in Liquid nitrogen, then store at -70 C. This will be used for potential send out tests for metabolic/genetic testing - 1 piece (similar in thickness to a match or a rounded toothpick) should go into glutaraldehyde for electron microscopy. You may receive 1container/piece Needle biopsies- ( See above information for clinicians): - Take a small piece (similar in thickness to a match or a rounded toothpick) and put in glutaraldehyde for Electron microscopy studies. - The remainder should be frozen following the Freezing procedure described below or in the links provided in Resources. 2

3 FREEZING PROCEDURE OF SKELETAL MUSCLE BIOPSIES FOR HISTOCHEMCIAL ANALYSIS -Confirm patient s name, medical record number, DOB. -Supplies are on the upper shelf of the muscle lab and first drawer labeled Freezing supplies. -Isopentane is in the 4C refrigerator -Liquid nitrogen is under the bench of muscle lab -Take tissue out. It may come in wet gauze. Transfer to a dry gauze and gently pad the tissue to remove extra moisture. Keep tissue away from cold air. Extra moisture and cold air can give freezing artifacts. Dry with gauze the container and place the container in the cryostat to cool down. This will be used to store the frozen tissue later on. -Take isopentane from 4 C refrigerator and cool down to -160C in Liquid nitrogen (LN) -While Isopentane cools down orient specimen in cross section 3

4 4

5 Clean instruments, return supplies to appropriate shelf and pour back left over isopentane in glass container and store at 4C for future use. Well done, you did it! Pictures courtesy of Dr. Limin Yu Acknowledgments: Thanks to THE MUSCLE TEAM! At the University of Michigan for working together in putting this guidelines together. 5

6 6