Standard Operating Procedure (SOP) For Rodent Stereotaxic Surgery
1.0 Purpose This Standard Operating Procedure (SOP) outlines the procedures for conducting stereotaxic surgery on rodents.
2.0 Responsibility Principal investigators (PIs) and their staff, veterinary care staff, or any individual performing stereotaxic surgery on rodents or assisting in those procedures are responsible for adhering to this SOP.
3.0 Materials
3.1. Anesthetics 3.2. Non-steroidal analgesic (such as carprofen or meloxicam; see Rodent Analgesia SOP) 3.3. Lidocaine-bupivacaine, 1:1 mixture (local analgesic) 3.4. Sterile 0.9% saline 3.5. Sterile ophthalmic ointment 3.6. Electric razor 3.7. Gauze 3.8. Antiseptic detergent (e.g., chlorhexidine 2% solution or povidone-iodine solution) 3.9. 70% alcohol 3.10. Heating disc, warming pad, or warm-water circulating pad (do not use electric heating pads), insulating material (thermal drapes, bubble wrap) 3.11. Sterile surgical instruments 3.12. Sterile gauze and swabs 3.13. Drill, sterile stainless steel screws, sterile cannulae 3.14. Suture material or wound clips (Autoclips) 3.15. Dry bead sterilizer or cold sterilization agents (e.g., glutaraldehyde) and 70% alcohol (as a rinsing agent).
4.0 Procedures
4.1 Pre-operative Preparation
4.1.1. Perform pre-operative procedures at a safe distance from the surgical environment to prevent contamination with hair.
4.2 Anesthesia and Pre-surgical Care
4.2.1. Anesthetize the animal according to the Rodent Anesthesia SOP. 4.2.2. Apply ophthalmic ointment in both eyes to prevent corneal desiccation. Reapply as needed. 4.2.3. Administer a non-steroidal anti-inflammatory analgesic (such as carprofen or meloxicam as per rodent analgesia SOP). 4.2.4. Administer 0.2 to 0.5 mL/10g body weight of 0.9% sterile saline subcutaneously. 4.2.5. Shave the top of the head and remove hair. 4.2.6. Secure the animal in the stereotaxic frame. 4.2.7. Place a heat source under the animal or wrap the animal with insulating material (e.g., thermal drapes, bubble wrap). 4.2.8. Wash the surgical site with 2% chlorhexidine solution or povidone-iodine solution. Be careful not to wet the animal. 4.2.9. Apply 70% alcohol with gauze or swabs to the head. Be careful not to wet a large area on the animal as the evaporation of alcohol will lead to heat loss. 4.2.10. Repeat steps 4.2.9 and 4.2.11 twice.
4.3 Surgical Procedure
4.3.1. Ensure that all the necessary materials are at hand. 4.3.2. Begin surgery with clean and sterile surgical instruments, handling them aseptically. 4.3.3. Designate a sterile area on the working surface for sterile materials (instruments, suture material, drapes, gauze, etc.). 4.3.4. Prior to surgery, verify the depth of anesthesia by loss of the animal's pedal withdrawal (toe pinch) reflex. 4.3.5. Expose the cranium by making a surgical anterior-posterior incision with a scalpel blade (for lesions) or by cutting a circular fold of skin with scissors (for cannula placement). 4.3.6. Avoid contact of tissues with fingers by using the tip of instruments. 4.3.7. Apply local anesthetics (mixture of bupivacaine and lidocaine) on the periosteum. 4.3.8. Reflect the skin, scrape the skull, and wipe the skull surface with sterile swabs to remove blood. 4.3.9. Using a hand-held drill, drill burr holes (up to 2 mm in diameter for rats and 1 mm in diameter for mice) into the skull for the insertion of stainless steel securing screws. 4.3.10. Insert stainless steel screws. 4.3.11. Make a craniotomy over the target area of the brain.4.3.12. Guide cannulae, temporarily fitted with internal cannulae, are inserted into the target area according to appropriate stereotaxic coordinates.
4.3.13. Dental acrylic is applied around the cannulae, screws, and any exposed cranium to secure the cannulae in place. 4.3.14. When the cement hardens, the internal cannulae are replaced with "dummy" cannulae (obturators), which are inserted into the guide cannulae to maintain patency. 4.3.15. A plastic dust cap is placed to protect the cannula assembly. 4.3.16. Infiltrate the wound with a local anesthetic, e.g., the mixture of lidocaine and bupivacaine, prior to closing the skin. Refer to Rodent Analgesia SOP. 4.3.17. Close the skin with polyamide-nylon, PDS, or Vicryl; size: 3-0 or 5-0, or the incision can be closed using wound clips (Autoclips); 7mm or 9mm. Sutures or staples must be removed after 10 days. 4.3.18. Disinfect the instruments between each animal by dipping them in a hot glass bead sterilizer for approximately 30 seconds after removing any blood or debris (let cool completely) or in a liquid sterilizing solution (e.g., glutaraldehyde or equivalent) for >5 minutes and then rinse with 70% alcohol. For liquid sterilization, it is recommended to use two alternating surgical kits to increase contact time with the solution. 4.3.19. Dip suture material in 70% alcohol between each animal.
4.4 Surgical Monitoring and Supportive Care
4.4.1. Refer to Rodent Surgery SOP.
4.5 Post-operative Care
4.5.1. Refer to Rodent Surgery SOP. 4.5.2. Repeat the administration of a non-steroidal anti-inflammatory analgesic (such as carprofen or meloxicam as per rodent analgesia SOP) every 24 hours for 1 to 2 days.
END OF THE DOCUMENT
You may like to read these links:
1. List of All SOPs and Documents for the Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for the In-vivo Laboratory.
4. List of All SOPs and Documents for Clinical Research
5. List of Instruments and Equipment Used In Laboratory Animal Facility
6. Animal Facility Design - Small Laboratory Animals (Rat, Mice, Rabbit, Guinea Pig)
7. List of Chemicals Needed for In-vivo Laboratory
8. List of Physiological Data for Small Laboratory Animals
9. List of All SOPs and Documents for the Animal House Facility
Hi. I’m Writer of Researchsop.com. ’ ’ Please share these SOPs to all concern pharma people for their development. I like to fullfill the need of curious people. These things inspire me to make things looks better.
0 comments:
Post a Comment