Standard Operating Procedure for performing a muscle biopsy Effective date: 31.10.2016 Review due date: 30.08.2018 Original Author Name: Richard Metcalfe and Abdullah Alghannam Date: 14.12.2012 Position: Ph. D. Students Reviewer Name: Ben Lee Signature: Position: Physiology Lab Technician Date: 31.08.2016 Approved by Name: Prof Keith Stokes Signature: Position: Deputy Head of Department Date: 31.10.2016 Amendment Chronology Version number Effective date Reason for amendment Reviewed/Amended by 1 14.12.2012 First issue Richard Metcalfe and Abdullah Alghannam 2 31.10.2016 No amends Ben Lee 3 27.11.2017 Minor changes to procedure and autoclaving. Appendices removed. Mark Thomas Pippa Heath Version: 2 Effective date: 31.10.2016 1
Performing a Muscle Biopsy Safety Information: Prior to performing this procedure please refer to the following risk assessment: Purpose: muscle_biopsy The purpose of the muscle biopsy technique is to obtain a sample of muscle tissue for research purposes. Decontamination of biopsy equipment involves the destruction or removal of any micro-organisms present and is necessary to prevent cross-contamination or infection of individuals. Cleaning, disinfection and sterilisation are used in the decontamination process to maintain sterility, integrity and prolong the life-span of the equipment. Contents: Only individuals who have undergone the required training, and read and understood the risk assessment, may perform this procedure. Best practice must be adhered to at all times. 1. Equipment Required 2. Preparation 3. Taking a sample 4. Decontamination and sterilisation 5. Ordering Information Equipment Required: Materials for Performing the Biopsy Scalpel and Blade (Swann morton, stainless steel, size 11) Gauze Converter Sterile Gloves Biopsy (Bergstrom) Needle Anaesthetic (Lidocaine) Iodine (Videne) & Iodine reservoirs Steri Strips 2 x 5ml syringe 100ml syringe Version: 2 Effective date: 31.10.2016 2
Catheter tubing Needle (orange) or grey 27 2 x Needle (green) 21 Forceps Sterile Scissors Regular Scissors Coban Tape Sharps bin Hazardous waste bin Liquid nitrogen canister & liquid nitrogen Razors Autoclave bags Spatula Adhesive Island Dressings (Primapore) Eppendorf safe-lock tubes. Medical tweezers (fine with curved tips) Stitches Small metal bowl for liquid nitrogen (optional) Materials for Decontamination Bergstrom Biopsy Needles Metal Scissors Metal Forceps Sterilising Pouch Autoclave Indicator Strip Autoclave Validation Vial (Sterikon Plus Bioindicator VWR Order number 1102740002) Ultrasonic Bath (2.5 L capacity) Basket for biopsy equipment Sonozyme Enzymatic Detergent Milton Sterilising Solution Ultra-pure water Wire Brush Preparation: 1. Sterilised biopsy needle in autoclave bag 2. Fill canister with liquid nitrogen 3. Attach catheter to 100ml syringe in preparation for needle. Cut the tube in a sharp angle. 4. With green needle poke 3 holes in top of eppendorf ready for liquid nitrogen + sample Taking a sample: 1. Prepare a sterile tray by placing a sterile sheet. Version: 2 Effective date: 31.10.2016 3
2. Place the reservoir and Videne on the sterile tray. 3. Shave the lateral aspect of the thigh to prepare the biopsy area. 4. Request subject to tense their quadriceps to establish the anatomical location for the incision (approximately 45 o from standing). 5. Sanitise hands, put sterile gloves on 6. Apply Videne on the entire visible area of the thigh through an outward circular motion for 30 seconds. Allow to dry naturally and mark site with fingertip 7. Load two 2.5ml syringes with anaesthetic (Lidocaine), expel air bubbles outside bottle. Without touching either needle, place a grey 27 needle and a green 21 needle onto each of the syringes. 8. Insert the grey needled syringe in a shallow angle to penetrate the skin; administer the anaesthetic upwards towards the (circle as a fan shape). 9. Insert the green needled syringe vertically in one motion (relatively quickly) to feel the facia (subtle increase in resistance should be felt); Continue to administer the anaesthetic on the facia (circle as a 2 pence piece). 10. Wait for 2 minutes. 11. Ensure that the subject has no pain sensations before commencing to the next step (confirm with the subject). 12. Insert blade vertically, and cut along skin to reach the fascia. Continue with a backward and forward motion, while twisting the blade in the end of the incision to allow the opening of the facia. 13. Flood with Videne and place pressure with gauze on top 14. Open biopsy needle autoclave bag 15. Attach needle to 100ml tube by the catheter tube. 16. Insert biopsy needle vertically, twisting as you insert and locate notch in facia 17. Apply some pressure by moving laterally from the femur. Ensure that the window in the needle is also facing laterally. 18. Expel air from needle and hold horizontally before performing suction. 19. Pull needle up, pull syringe out and snip into muscle (2-3 times). Rotate 1/8 turn in between each snip. 20. Remove needle and flood wound area with Videne and place pressure with gauze on top. 21. Use one stitch to close the wound. 22. Seal wound area with steri-strip and place Adhesive Island Dressing. 23. Apply pressure with gauze for at least 5 minutes. Wrap with Coban tape for pressure. Version: 2 Effective date: 31.10.2016 4
24. Decant muscle into eppendorf of liquid nitrogen. Participant care: Instruct the participant on maintenance of biopsy site and how to prevent infection. The stitch will need to be removed in 5-7 days. Coban tape needs to remain on for a minimum of 2 hours, but ideally 24 hours to reduce bruising risk. Provide them with extra Primapore. Decontamination and Sterilisation: Materials Bergstrom Biopsy Needles Metal Scissors Metal Forceps Sterilising Pouch Autoclave Indicator Strip Autoclave Validation Vial (Sterikon Plus Bioindicator VWR Order number 1102740002) Ultrasonic Bath (2.5 L capacity) Basket for biopsy equipment Sonozyme TM Enzymatic Detergent Pre-sept Sterilising Solution Ultra-Pure Water Wire Brush Procedure: 1. Wash all biopsy equipment: Bergstrom needles (all component parts are separated and cleaned individually) in warm water and physically remove any organic matter, blood and other deposits with a soft nylon bristled brush. Ensure the brush is cleaned with detergent and hot water directly after use. 2. Place the items in an ultrasonic bath containing a solution of 0.3% Sonozyme enzymatic soap (6.25mls per 2 litres of water) and sonicate for 10-15 minutes at 30 o C. If decontaminating a large batch of biopsy equipment, a maximum of five items should be placed in the basket and treated at any one time. Make sure items do not overlap in the Sonicator. 3. Rinse all items in water. 4. Soak the items in a suitable disinfectant i.e Pre-sept sterilising solution for fifteen minutes and rinse in ultrapure water. Version: 2 Effective date: 31.10.2016 5
5. Dry items with a blue towel and place individually in sterilising pouches. Seal with autoclave tape and label the tape with the item name, Health, the lab, your initials and the date in permanent ink. 6. Place in Biology autoclave room on dry shelf. End of Document Version: 2 Effective date: 31.10.2016 6