ZEBR

1. ZEBR

2. AFISH

3. HOLDI

4. NG

5. CHAM

6. BER

7. Team Members: Austin Ramme, Kristen Sipsma,

8. Mike Haggerty, Andrew Neumann Client:

9. Jill Kolesar, Pharm.D., BCPS Advisor: Professor Justin Williams

10. Abstract Our objective is to build an apparatus to house between one and three zebrafish during a diagnostic micro CT scanning process. The device will be used for an evaluation of carcinogenesis in a zebrafish model research project currently being conducted at the UW hospital. The ultimate goal of this research is to cure cancer using drugs which eliminate the blood supply to malignant tumors. To guarantee the best imaging quality, the device must be composed of materials compatible with the micro CT scanner and must immobilize the fish. The zebrafish must remain alive for the duration of the twenty minute imaging process.

11. Project Motivation • Client’s Objective • An Evaluation of Carcinogenesis in Zebrafish Model • Ultimate Goal: Cure Cancer by Eliminating Malignant Tumors • Client’s Device • Cardboard • Zebrafish Slits • High Mortality Rate Client’s Current Device

12. 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

13. Design Specifications • Max Dimensions: • Width- 4 in • Length- 1 ft • Height- <4 in • No Leakage • Zebrafish Must Survive • Metal, Glass, and Dense Materials Must Not Be Used • Minimize All Motion of the Zebrafish • Disposable or Autoclavable Micro CT Scanner

14. Testing: Phase 1 Thirty Minute Bob Date: February 11, 2004 Purpose: Determine Zebrafish Survival Rate in Cuvette Procedure: Leave Fish in 3.5 ml Cuvette for 30 min Conclusions: Fish Can Survive in 3.5 ml Cuvette for 30 min No Adverse Behavior in Cuvette Fish Preferred Horizontal Orientation

15. Testing: Phase 2 • Dates: April 4, April 6, April 12, 2004 • Purpose: Test Restraints in Cuvette • Procedure: • Test Various Restraints in Cuvettes for 20 min 1. 1.5 ml semi-micro cuvette with dense dish scrubber 2. 1.5 ml semi-micro cuvette with low-density tile sponge 3. 3.5 ml cuvette with low-density tile sponge • Conclusions: • 75% Survival Rate with Restraint 1 • 100% Survival Rate with Restraint 2 and 3 • Restraint 2 and 3 Equally Immobilized the Fish

16. Final Design: Prototype I • Calculated Base • Cuvette Apparatus • Allows 1-7 Zebrafish Standard Cuvette & Sponge Vs. Semi-Micro Cuvette & Sponge

17. Prototype I: Micro CT Testing Semi-micro Cuvette Flanked by Two Standard Cuvettes

18. Final Design: Prototype II • Calculated Base • Micro CT Compatible • Allows 1-3 Zebrafish • Standard Cuvette Design for Maximum Immobility Standard Cuvette & Sponge

19. Prototype II: Micro CT Testing Three Dimensional View of Three Standard Cuvettes

20. Manufacturing • Cut the Plexiglas • Table Saw • Band Saw • Milling Machine • Gluing • Finish Sanding • Testing

21. Materials and Costs

22. System Advantages • High Survival Rate • Immobilizes Zebrafish • Compatible with Micro CT Scanner • Disposable Design • Easy Maintenance • No Cross-Contamination • Flexible 1-3 Zebrafish Design

23. Future Work • Individually Scan Sponge • Keep Fish Alive for Extended Periods of Time • Determine How to Further Reduce Motion • Find Better Method of Inserting Fish and Sponge into Cuvette

24. References • Aquadex. 19 Feb 2004 <http://aquadex.info/fish/Danio_rerio.cfm>. • 2. Bailey, George S. Research. Oregon State University Molecular and Cellular Biology Program. 06 Feb 2004. <http://www.cgrb.orst.edu/mcb/faculty/bailey/>. • 3. eXplore RS Micro CT System Pre-clinical Imaging. GE Medical Systems. 08 Feb. 2004. <http://www.gemedicalsystems.com/rad/nm_pet/preclinicalimaginghome.html>. • 4. Functional Imaging: Pre-clinical Imaging. GE Medical Systems. 15 Feb. 2004. <http://www.gemedicalsystems.com/>. • 5. How Fish Breath?. Pet Place. 07 Feb 2004. <http://petplace.netscape.com/Articles/ artShow.asp?artID=3980 • 6. Info Center. Duke University. 30 Jan 2004. <http://wwwcivm.mc.duke.edu/zebra /info.html> • 7. Keil, Eli. Fish Base. World Wide Web Electronic Publication. 19 Feb 2004. <http://www.fishbase.org/Summary/SpeciesSummary.cfm?genusname=Danio&speciesname=rerio>. • 8. McAllister, Michael Jason. Zebrafish: An Integrated 3D Magnetic Resonance Study of Brachydanio Rerio. Duke Medical Center. 30 Jan 2004. <http://wwwcivm.mc.duke.edu/zebra/info.html>. • 9. McMaster-Carr. 21 April 2004. <http://www.mcmaster.com/>. • 10. Perfector Scientific. 21 April 2004. <www.perf-sci.com>. • 11. Robinson, Tracy and Bates, Todd. Thousands of fish die in Belmar's Silver Lake. Asbury Park Press. 30 Jan 2004. <http://www.app.com/app2001/story/0%2C21133%2C 777232%2C00.html • 12. Sprague, Judy et al. The Zebrafish Information Network: a resource for genetic, genomic, and developmental research. The Zebrafish International Resource Center, University of Oregon. 30 Jan 2004. <http://www.cs.uoregon.edu/~douglas/PAPERS%20DOWNLOAD/Sprague01.pdf>. • 13. TeraRecon to Demonstrate High Performance Cone Beam CT Reconstruction Server at RSNA. TeraRecon Inc. 9 Feb 2004. <http://www.rtviz.com/news/press/2001/11_november_13_pr.html>. • 14. Vascular Anatomy of the 3.5-4 dpf Zebrafish Embryo. The Interactive Atlas of Zebrafish Vascular Anatomy. 20 Feb. 2004. <http://mgchd1.nichd.nih.gov:8000/zfatlas/FinalDesign2/L26.html>. • 15. Zebrafish Cancer. Howard Hughes Medical Institute, Boston Children’s Hospital. 06 Feb 2004. <http://zon.tchlab.org/zebrafishcancer.htm>. • 16. Zebrafish Research: Activities at the NIH. Trans-NIH Zebrafish Initiative. 30 Jan 2004. <http://www.nih.gov/science/models/zebrafish/reports/activities.html>.

25. A Special Thanks • Professor Justin Williams – Advisor • Dr. Jill Kolesar – Client • Dr. Jamey Weichert – Micro CT Expert • Dr. Marc Longino – Micro CT Expert