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Muscle Biopsy, Skeletal, Fresh Tissue

Skeletal Muscle Biopsy - Fresh Tissue, Muscle Biopsy, Skeletal

Department

Histology

Instructions

• Beaumont Laboratory strives to provide preliminary results via a phone call regarding presence or absence of inflammation or vasculitis, and discuss other relevant findings with the patient's Neurologist, Rheumatologist, Internal Medicine physician, Pediatric physician, or other clinician within one to two business days of receiving the biopsy. This service can only be provided if the proper contact information is provided in the submitted paperwork.
  
  
• The specimen must be accompanied by:

Outreach
Muscle Biopsy, Beaumont Laboratory Outreach
Requisition 5728 031910 OS8.

• Electronic Ordering with generation of paper requisition: EPIC Optime/Navigator order “LAB6798 Surgical Pathology Tissue”
  
  
• Each specimen requisition must include patient's full name, date of birth, patient's social security number (only last 4 of SSN), hospital number including visit number, authorizing physician and/or clinician name, specimen source including identifiers and pre-op diagnosis. For proper patient care and pathologic examination, the name and phone number of the Neurologist and/or Rheumatologist is required.
  
  
• Follow-up on cases is always appreciated. Please contact Beaumont Laboratory by call 800-551-0488, Option 5, to comment on your service. Thank you for allowing Beaumont Laboratory to participate in the care of your patient(s).
  
  

Royal Oak

• Royal oak Surgical Pathology/Histology laboratory processes all muscle biopsies
  
  
• Surgery for muscle biopsies needs to be scheduled in the day so that the muscle biopsy is RECEIVED in Royal oak Surgical Pathology/Histology Laboratory prior to 3:00 pm, Monday – Friday. Muscle biopsies should not be performed on weekends.
  
  

Non Royal Oak acute care campuses (Dearborn, Troy, Farmington Hills, Grosse Pointe, Taylor, Trenton, Wayne)


• Please notify the histology laboratory 24 hours in advance by calling (see below) during normal hours of operation. The laboratory will notify Royal Oak of pending biopsy.
  
  
• Muscle biopsies should be scheduled between the hours of 7:00 am and 12:00 pm to allow for time needed to transport specimen to Royal Oak for processing.
  
  
• Muscle biopsies should be placed in a muscle clamp if possible and gently wrapped in saline moistened gauze and placed in a sterile container with all necessary patient information including source and site of biopsy. DO NOT PLACE BIOPSY IN FORMALIN.
  
  
• Please include "surgical specimen requisition/order" form with necessary patient information to include the source and site of specimen.

Specimen Collection Criteria

Collect: Biopsy specimen collected as described below:

• Option 1: Specimens at Beaumont Health, Royal Oak: Sharply dissect a portion of skeletal muscle, measuring at least 2.0 x 1.0 x 1.0 cm, gently wrapped in saline - moistened gauze (not floating in saline as this causes freezing artifact), placed into an air tight specimen container and delivered to Anatomic/Surgical Pathology immediately. It is preferable, but not mandatory, to place the biopsy in a muscle clamp. The specimen container must be labeled with the patient's full name (first and last), date of birth, hospital number including visit, specimen source and site (for example "left quadriceps muscle").
  

• Option 2: Hospitals Close to Beaumont Health, Royal Oak: For specimens to be received within 2 to 3 hours after the time of biopsy. If possible, muscle biopsies should preferably be placed in a muscle clamp, gently wrapped in a saline-moistened gauze, and placed into an air-tight specimen container. The air-tight container is placed within a cooler on "wet-ice" or cooling pack (NOT Frozen), and delivered to Beaumont Hospital Anatomic/Surgical Pathology Laboratory as a STAT delivery.
  
  
• Option 3: Hospitals Far from Beaumont Health, Royal Oak: For specimens to be received than three hours after the time of biopsy. Triage and prepare the tissue for subsequent evaluation. This entails dividing the muscle biopsy into 4 portions with a new scalpel or razor blade.
  
  
  • Portion #1 - Snap Frozen for Enzyme Histochemistry: The frozen portion (approximately 1.0 cm in length) should be mounted upright for a cross-sectional cut on a chuck or cork-disc with OCT. Leave a large portion of the tissue to be cut above the mounting medium. DO NOT cover the entire muscle biopsy with OCT or talcum powder. Freeze the specimen for 45 to 60 seconds in isopentane/isobutene which has been pre-cooled to at least -160°C with liquid nitrogen. Store the specimen at -70°C until ready to ship. Ship on dry ice.
    
    
  • Portion #2 - for Electron Microscopy (EM): Sharply dissect 5-7 mm long, 2-3 mm wide longitudinal strips of muscle from a well oriented and preserved area of the muscle. Gently stretch these onto a piece of cardboard/index card or a portion of tongue depressor and place in buffered glutaraldyhyde (2-5%). The specimen should be refrigerated but NOT FROZEN.
    
    
  • Portion #3 - for Formalin Fixation and Routine Processing: If you are convinced this is not a metabolic myopathy, submit the remainder of tissue in formalin (10% neutral buffered formalin).
    
    
  • Portion #4: If any possibility of metabolic myopathy or genetically inherited disorder is suspected, freezing an eraser-sized portion of tissue for biochemical and/or genetic studies is warranted. This is best accomplished by placing the tissue in a small plastic bag (without OCT or talcum) which is then cooled in a -70° freezer, but may also be accomplished with dry ice or liquid nitrogen. This sample needs to be pure muscle, without adipose or fascia.
    
    
• Other: A portion of skin (at least 0.5 cm in size) obtained under sterile conditions and submitted in tissue culture media is suggested in pediatric metabolic cases, for submission to Cytogenetics for fibroblast culture and cryopreservation. This allows possible future additional genetic analysis should there be insufficient muscle tissue to pursue all the desired studies. The skin should not be frozen and should be sent under ambient conditions in tissue culture media.

Physician Office/Draw Specimen Preparation

• Hospitals close to Beaumont Health should deliver the fresh specimen immediately to Anatomic /Surgical Pathology via a courier/transportation service chosen by the facility sending the specimen. STAT transportation can be arranged by calling Beaumont Laboratory Outreach 800-551-0488. The specimen should always be handed to one of the surgical pathology assistants.
  
  
• Hospitals far from Beaumont Health need to send the specimen by express courier to the Department of Anatomic/Surgical Pathology following OSHA regulations. The frozen portion of the sample needs to be shipped on dry ice. The formalin and glutaraldehyde fixed tissue should be shipped at room temperature (20-26°C or 68-78.8°F).
  
  
• If the specimen is being shipped via express courier, then it is wise to track the specimen (via tracking number) until it arrives at the intended destination (occasional specimens have been inadvertently misrouted by the courier and received in the Laboratory later than intended in a thawed state). To avoid this, notify Surgical Pathology when sending a muscle biopsy and ensure that the frozen tissue specimen is shipped with an adequate amount of dry ice.

Preparation for Courier Transport

Transport: Biopsy specimens prepared as described above, at room temperature (20-26°C or 68-78.8°F). Specimens are hand delivered to Surgical Pathology.

Rejection Criteria

• There are no rejection criteria.
• Poor freezing technique will hamper evaluation.
• Delay in processing of the fresh muscle tissue will affect the staining and diagnosis.
• Any missing information will also delay processing and final diagnosis.

Inpatient Specimen Preparation

Fresh muscle biopsies should always be delivered immediately to Surgical Pathology and should be handed to one of the Surgical Pathology assistants. NEVER leave a fresh muscle biopsy in the specimen pick-up area.

In-Lab Processing

Muscle biopsy is immediately grossed into 3 or 4 portions, depending on the clinical history. One portion is snap frozen for histochemical studies. The second portion is placed into glutaraldehyde for Electron Microscopy (EM). The third portion is placed into formalin and processed by Histology. The fourth portion is frozen for possible future genetic and/or biochemical studies. Tissue triage may vary depending on the amount of tissue available and the clinical context.

Storage

Specimen Stability for Testing:

Specimens are processed immediately upon receipt in the Laboratory.

Specimen Storage in Department Prior to Disposal:

After case is signed-out: Hematoxylin and eosin stained sections of formalin-fixed tissue and frozen tissue can be sent to the submitting hospital's Pathology Department. The remainder of the slides, paraffin blocks, snap frozen tissues, EM blocks, grids and photomicrographs are retained at Beaumont Health, Royal Oak.

Laboratory

Royal Oak Surgical Pathology/Histology Laboratory

Performed

Monday – Friday, 6:00 am – 3:00 pm.
Final results typically are available within 1 week for inflammatory cases, 1 week to 2 weeks for metabolic and muscular dystrophy. Very complex cases may take longer.

Test Methodology

• Routine Processing with Hematoxylin and Eosin stained slides prepared from frozen and formalin fixed tissue.
• Special Staining Procedures: Masson trichrome, PAS with and without diastase, Congo-red, Oil-red-O.
• Enzyme Histochemistry: Esterase, alkaline phosphatase, myophosphorylase, NADH (Nicotinamide Adenine Dinucleotide, reduced), cytochrome oxidase, succinic acid dehydrogenase, acid phosphatase.
• Electron Microscopy: Methylene Blue Azure-stained semi-thin sections, ultrastructural study.
• Immunoperoxidase: Markers for inflammatory myopathy and muscular dystrophy usually Dual Myosin Muscle Frozen, CD3, CD163, MHC and MAC. 

Interpretation

Microscopic diagnosis of prepared slides and ultrastructural photomicrographs.

Contacts

Last Updated

5/30/2023