1. A Zebrafish Holding Chamber for Use with a Micro CT Scanner

3. Team Members Austin Ramme, Kristen Sipsma, Andrew Neumann, and Mike Haggerty Client Jill M. Kolesar, Pharm.D., BCPS Associate Professor of Pharmacy Advisor Professor Justin Williams

4. Presentation Outline  Problem Statement  Background Information  Current Devices  Design Specifications  Alternative Designs  Final Design  Future Work

5. Problem Statement  Client –An Evaluation of Carcinogenesis in a Zebrafish Model –Ultimate Goal: Cure Cancer  Design Team –Design a Zebrafish Holding Chamber –Compatible with a Micro CT Scanner –Sustains Zebrafish Life

6. Background Information  CT = Computed Tomography  Benefits of Zebrafish Research –Quick Breeders –Low Maintenance –Mapped Genome  Zebrafish Anatomy –Length: 1-6cm –Lifespan: 2 years –Water Temp: 64-75° F –Bony Fish: Breathe Without Moving Typical CT Scanner

7. Current Devices  Market Products –Rodents –Zebrafish  Client’s Device –Cardboard –Zebrafish Slits –High Mortality Rate Rodent Tray Client’s Current Device

8. Preliminary Experimentation  February 11, 2004 Conclusions –Survival: 1.5 mL for >30 min –Increased Breathing Rates –Preferred Orientation –No Adverse Behavior in Cuvette –No Adverse Behavior with Restraint

9. Design Specifications  No leakage  Zebrafish must remain alive for 20 minutes  Metal, glass, and dense materials must not be used  Minimize all motion of the zebrafish  Disposable or autoclavable Micro CT Scanner

10. Design 1: Falcon Tube  Falcon tube filled with water with sponge inside  Small slits to hold fish  One to two fish per tube  Sponge absorption

11. Side View of Plate in Container 3D View of Container – No Plates In Place Water Thumb – Screws Design 2: Plexiglas Container  Multiple Fish  Adequate Water  Contamination 65mm

12. Design 3: The Cuvette  Cuvette  Stabilization of Fish  Alignment Tray  Disposable/Inexpensive  Water Supply  Space Efficient

13. Criteria: Cuvette Design Plexiglas Container Falcon Tube with Sponge Cost433 Ease of Use 332 Constructability534 Effectiveness: Immobilization 443 Effectiveness: Survival Rate 554 Totals211816 Design Matrix Each criterion was rated on a 0 to 5 scale with 5 being the best.

14. Future Work  Cuvette Tops –Rubber Stopper –Market  Sizing Difficulties  Opaque Materials  Build Prototype  Prototype Testing

15. Questions ?