ZEBR

AFISH

HOLDI

NG

CHAM

BER

Team Members:

Austin Ramme,

Kristen Sipsma,

Mike Haggerty,

Andrew Neumann

Client:

Jill Kolesar, Pharm.D.,

BCPS

Advisor:

Professor Justin Williams

AbstractOur 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.

Project Motivation

Clients ObjectiveAn Evaluation of Carcinogenesis in Zebrafish ModelUltimate Goal:Cure Cancer by Eliminating Malignant TumorsClients DeviceCardboardZebrafish SlitsHigh Mortality RateClients Current Device

Background InformationCT = Computed TomographyBenefits of Zebrafish ResearchQuick BreedersLow MaintenanceMapped GenomeZebrafish AnatomyLength: 1-6cmLifespan: 2 yearsWater Temp: 64-75 FBony Fish: Breathe Without Moving

Typical CT Scanner

Design SpecificationsMax Dimensions:Width- 4 inLength- 1 ftHeight-

Testing: Phase 1Date: February 11, 2004Purpose: Determine Zebrafish Survival Rate in CuvetteProcedure:Leave Fish in 3.5 ml Cuvette for 30 min

Conclusions:Fish Can Survive in 3.5 ml Cuvette for 30 minNo Adverse Behavior in CuvetteFish Preferred Horizontal OrientationThirty Minute Bob

Testing: Phase 2Dates: April 4, April 6, April 12, 2004Purpose: Test Restraints in CuvetteProcedure:Test Various Restraints in Cuvettes for 20 min1. 1.5 ml semi-micro cuvette with dense dish scrubber2. 1.5 ml semi-micro cuvette with low-density tile sponge3. 3.5 ml cuvette with low-density tile spongeConclusions:75% Survival Rate with Restraint 1100% Survival Rate with Restraint 2 and 3Restraint 2 and 3 Equally Immobilized the Fish

Final Design: Prototype IStandard Cuvette & SpongeSemi-Micro Cuvette & SpongeVs.Calculated BaseCuvette ApparatusAllows 1-7 Zebrafish

Prototype I: Micro CT TestingSemi-micro Cuvette Flanked by Two Standard Cuvettes

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

Prototype II: Micro CT TestingThree Dimensional View of Three Standard Cuvettes

ManufacturingCut the PlexiglasTable SawBand SawMilling MachineGluingFinish SandingTesting

Materials and Costs

ItemQuantityTotal PriceZebrafish12$9.00Tile and Grout Sponges1$5.97Plexiglas 2x1x1/41$4.00Acrylic Adhesive3$5.07Standard Cuvettes500$29.50Synthetic Steel Wool1$3.49Total$57.03

System AdvantagesHigh Survival RateImmobilizes ZebrafishCompatible with Micro CT ScannerDisposable DesignEasy MaintenanceNo Cross-ContaminationFlexible 1-3 Zebrafish Design

Future WorkIndividually Scan SpongeKeep Fish Alive for Extended Periods of TimeDetermine How to Further Reduce MotionFind Better Method of Inserting Fish and Sponge into Cuvette

ReferencesAquadex. 19 Feb 2004 .2. Bailey, George S. Research. Oregon State University Molecular and Cellular Biology Program. 06 Feb 2004. .3. eXplore RS Micro CT System Pre-clinical Imaging. GE Medical Systems. 08 Feb. 2004. .4. Functional Imaging: Pre-clinical Imaging. GE Medical Systems. 15 Feb. 2004. .5. How Fish Breath?. Pet Place. 07 Feb 2004.

A Special ThanksProfessor Justin Williams AdvisorDr. Jill Kolesar ClientDr. Jamey Weichert Micro CT ExpertDr. Marc Longino Micro CT Expert