A Zebrafish Holding Chamber for Use with a Micro CT Scanner

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

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

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

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

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

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

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

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

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

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

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

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

Questions ?