<p> Supplemental Guide</p><p>Instructor’s Supplemental Guide Supplemental Guide</p><p>The goal of this guide is to aid the lab instructor in setting up and running the rat dissection and lung casting laboratory procedure described in the student manual. It is meant to supplement the student manual, and to provide a list of the safety precautions, materials, and personnel needed to perform the procedure. Prior to starting the lab all students should read through the procedure and watch the instructional video. It also important that the instructor reviews basic lab safety guidelines and highlights the safety precaution listed in this manual.</p><p>2 Supplemental Guide</p><p>After reviewing basic lab safety guidelines, both the laboratory instructor and participating students should familiarize themselves with the following safety precautions:</p><p>Handling Live Rats</p><p>1. Thick leather or steel mesh gloves should be worn at all times when </p><p> handling rats.</p><p>2. Only pick up rats by tail and never leave un-caged rat unattended.</p><p>3. Always use restraining device when anesthetizing rat.</p><p>4. Make sure the restraining device is secured to the work surface.</p><p>Euthanization</p><p>1. Never leave heat lamp unattended when in use.</p><p>2. Use of Sodium Pentobarbital is only permitted under the supervision </p><p> or approval by licensed personnel in the RIT Biological Sciences </p><p>Department.</p><p>3. Take extra caution handling syringes used for Sodium Pentobarbital </p><p> injections, and be sure to dispose of syringes in designated sharps </p><p> receptacle.</p><p>3 Supplemental Guide</p><p>Handling Euthanized Rats</p><p>1. Wear rubber gloves at all times and clean up any spilled blood right </p><p> away.</p><p>2. Perform dissection in designated pan with a mat.</p><p>Dissection and Casting</p><p>1. Use extreme caution when working with surgical tools (scissors, </p><p> forceps, probes, etc.).</p><p>2. Ensure CO2 cylinder is secured to solid work surface.</p><p>3. Use caution when working with syringes for saline and casting </p><p> agent.</p><p>4. Never leave syringe pump unattended when in use.</p><p>5. Wear gloves when working with Silastic E casting agent and catalyst.</p><p>6. Monitor carbon dioxide’s pressure and the flow rate of both saline </p><p> and casting material periodically. Make sure there are no sudden </p><p> changes in either the gas pressure or flow rate.</p><p>Curing</p><p>1. Remove all surgical tools from dissection tray and clean with </p><p> caution.</p><p>2. Cover dissection tray and place in cool, safe location for a minimum </p><p> of 24 hours, but not to exceed 36 hours.</p><p>4 Supplemental Guide</p><p>Excision and Rat Disposal</p><p>1. Use extreme caution when handling NaOH and HCl solutions by </p><p> wearing gloves and avoiding all contact.</p><p>2. Never pour acidic or basic solution down drain. Always neutralize </p><p> before disposal, and be sure to double check pH with litmus paper.</p><p>3. Double wrap rat in plastic bag and label accordingly. Place in </p><p> designated freezer until disposed of appropriately (see personnel </p><p> contact list).</p><p>5 Supplemental Guide</p><p>1. Heat lamp</p><p>2. Scales</p><p> a) Triple beam balance (for rat).</p><p> b) Digital scale (casting agent).</p><p>3. Storage</p><p> a) Designated freezer for dead rats.</p><p> b) Lockable cabinet for Sodium Pentobarbital.</p><p> c) Steel cabinet for other chemicals.</p><p>4. Programmable syringe pump</p><p>5. CO2 cylinder</p><p> a) CO2 low pressure regulator.</p><p> b) ¼” NPT to 1/16” ID barbed urethane adapters.</p><p>6. Restraining device</p><p>7. Gloves</p><p> a) Latex rubber gloves in a variety of sizes.</p><p> b) Additional non-latex gloves in case of latex allergies.</p><p> c) Leather or steel mesh glove for handling rats.</p><p>8. Surgical tools</p><p> a) Scissors.</p><p> b) Forceps.</p><p>6 Supplemental Guide</p><p> c) Probes.</p><p> d) T-pins.</p><p> e) Suture string.</p><p>9. Dissection tray and lid cover</p><p>10. Clear 0.056”ID tubing</p><p>11. Plastic syringes</p><p> a) 5cc syringe with 18 gauge 1 ½” needle.</p><p> b) 2 cc syringe with 22 gauge ½” needle.</p><p>12. Silastic E and hardener</p><p>13. Degassed saline</p><p>14. 4M NaOH solution</p><p>15. 4M HCl solution</p><p>16. Litmus paper for testing pH</p><p>17. Plastic bags and tape for disposal</p><p>18. Mild detergent</p><p>7 Supplemental Guide</p><p>RIT Biology Department: Dr. Thomas Frederick Office: 08-2102 Phone: (585) 475-2496 Email: [email protected]</p><p>RIT Mechanical Engineering Department: Dr. Risa Robinson Office: 09-2041 Phone: (585) 475-6445 Email: [email protected]</p><p>Environmental Safety Officer Helen Revelas Office: 04-A122 Phone: (585) 475-2040 Email: [email protected]</p><p>RIT Science Department: Glenn Robinson Office: 08-A224 Phone: (585) 475-5759 Email: [email protected]</p><p>8 Supplemental Guide</p><p>Syringe pump and dissection aids: Braintree Scientific</p><p> www.braintreesci.com</p><p>Bio lab supplies: Ward's Scientific www.wardsci.com</p><p>Pressure regulator parts: McMaster-Carr www.mcmaster.com</p><p>Silastic E: Dow Corning Inc. www.dowcorning.com</p><p>9</p>