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Transonic Systems Inc.

Measurement of Cardiac Output in the Mouse. Implantation on the Ascending Aorta Required Equipment Surgical Protocol Measurement Data References . Transonic Systems Inc. Required Equipment. Stereo Microscope Isoflurane Anesthesia Mouse Endotracheal Tube Rodent Respirator

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Transonic Systems Inc.

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  1. Measurement of Cardiac Output in the Mouse Implantation on the Ascending Aorta Required Equipment Surgical Protocol Measurement Data References Transonic Systems Inc.

  2. Required Equipment Stereo Microscope Isoflurane Anesthesia Mouse Endotracheal Tube Rodent Respirator Microsurgical Instruments Transonic 1.5PSL Flowprobe Negative Pressure System Flowprobe Button Transonic Flowmeter

  3. Transonic 400-Series Flowmeter 1.5PSL Flowprobe: designed for mouse ascending aorta Transonic Systems Inc.

  4. Mouse Surgical Tool Kit 1 - Vessel Cannulation Forceps 2 - # 5 45° angle microblunted, atraumatic Dumont 3 - D-aZ S & T vessel dilator forcep 4 - # 5 45° angle microblunted, atraumatic Dumont slightly altered very blunt tips 5 - Foerster octagonal curved - no teeth 6 - Foerster octagonal curved - with teeth 7 - Foerster octagonal straight - no teeth 8 - Goldstein Micro dissecting Retractors 3 x 3 prong (altered to 2 x 2 prong) 9 - Kleinert-Kutz microclip applying forceps 10- Schwartz micro-serrefines 11- Chest Tube 12 - Fine Iris Scissors w/ oversize fingerloops blunt/blunt 13 - Spring Scissors with fine small blades 14- Extra delicate mini-Vannas 15 - Halsey Micro Needle Holder extra light & delicate 16 - Castroviejo with lock and TC jaws 3 4 1 2 5 6 7 10 11 9 8 12 16 15 14 13

  5. Surgical Protocol: Implantation of a 1.5PSL Flowprobe on the Ascending Aorta heart Ascending Aorta After the mouse has been anesthetized and properly ventillated on a respirator, a right thoracotomy is performed in the 3rd intercostal space. Closure sutures are placed in the muscle layers at the beginning of the procedure. A modified rib retractor is inserted to visualize the heart and ascending aorta.

  6. Aorta Careful dissection along the ascending aorta is performed by blunt dissection using microsurgical vessel dilators to free the vessel from connective tissue. The aorta has been isolated from the pulmonary artery. Gently lift under the aorta to pass surgical silk under the vessel to aid in placing the vessel in the lumen of the probe.

  7. Aortic silk Note the surgical silk around the ascending aorta at the base of the heart. The retractors are removed and the 1.5PSL flowprobe is introduced into the thoracic cavity.

  8. Stabilizing suture The aorta is gently lifted into the opening of the flowprobe. Confirm that the vessel is within the probe by gently rotating the probe. Remove the surgical silk from around the aorta to prepare for closure. The cable is rotated laterally to the right and the thoracotomy closed over the probe. Reapproximate the muscles. A suture is passed around the flowprobe cable to stabilize the probe in position.

  9. Final Closure Procedures Chest Tube A chest tube attached to a negative pressure system can be monitored for bubbles indicating pneumothorax. Close the skin incision. For chronic monitoring, the probe connector is passed under the skin to the midscapular area by creating a subcutaneous tunnel using a pair of straight hemostats and gently grasping the CA4S connector. Dacron mesh is placed under the skin at the connector and the incision closed. A button cuff is installed over the connector and sutured in place through the dacron mesh.

  10. The mouse can be monitored for cardiac output after recovery in 3 - 5 days.During this time, the flowprobe will encapsulate in fibrous tissue to provide good signal transmission.

  11. Chronic Cardiac Output in Mice Transonic 1.5PSL probe on Ascending Aorta probe implanted 5/22/02 Attached to Dragonfly swivel for recording 2/21/03

  12. Continuous Conscious Cardiac Output in Mice Recording over ~ 8 days with 1 interruption 0.1 Hz; 10 samples/sec Note circadian cycles in flow. 2.25.03 9:30:00 2.5 days interruption to recording

  13. Conscious Cardiac Output in Mice Transonic 1.5PSL probe implanted 5/22/02 recording 2/25/03 Flow 14.2 ml/m Heart Rate 510 bpm SV = 28 µl

  14. 8 Minute Recording Of Cardiac Output Dramatic variability demonstrates dynamic range of mouse CO related to activity which cannot be recognized under acute anesthetized protocols. Walking CO = 19.3 ml/m HR = 675 bpm SV = 29 µl Resting Quietly CO = 8.97 ml/m HR = 405 bpm SV = 22 µl Walking CO = 20.04 ml/m HR = 600 bpm SV = 33 µl Sniffing CO = 18.1 ml/m HR = 630 bpm SV = 29 µl Walking CO = 10.5 ml/m HR = 428 bpm SV = 25 µl Walking at one end of cage Pushing bedding Walking to other side of cage Quiet Sniffing Quiet 02.27.03

  15. Equipment list Flowprobe: MC1.5PSL-JN-WM35-CA4S-GC Transonic Systems, Inc. Flowmeter: Transonic TS420 Flowmeter Module Skin Button Cuff: Transonic Systems, Inc. www.transonic.com Surgical Tools & Retractors: Fine Science Tools www.finescience.com Mersilene Mesh & Vicryl Sutures: Ethicon www.ethicon.com Electrical Swivel: Dragonfly Research & Development, Inc. www.dragonflyinc.com

  16. Transonic Systems Inc. Transonic Systems, Inc. gratefully acknowledges the assistance of the collaborators in the development of this protocol: • Thomas L. Smith, Ph.D. • Dept. of Orthopaedic Surgery • Wake Forest University Health Sciences • Michael F. Callahan, Ph.D. • Dept. Physiology & Pharmacology • Wake Forest University Health Sciences • Produced by: Margo Sosa • Senior Product Manager • Transonic Systems, Inc. • 34 Dutch Mill Road • Ithaca, NY 14850 • www.transonic.com

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