GXP-1702 Design of a Universal Microscope Incubator for Drug Screening of 3D Models of Engineered Myocardium A Major Qualifying Project Report Submitted to the Faculty of Worcester Polytechnic Institute In partial fulfillment of the requirements for the Degree of Bachelor of Science by Rachel Connolly _______________________ Marianne Kanellias ________________________ Heather Stratica ________________________ April 2016 Approved by: Professor George Pins, Advisor ______________________________ Professor John Sullivan, Advisor ______________________________ i Contents Abstract ........................................................................................................................................................ iv Authorship ..................................................................................................................................................... v Acknowledgements ...................................................................................................................................... vi Abbreviations .............................................................................................................................................. vii Table of Figures ........................................................................................................................................... viii Table of Tables ............................................................................................................................................. xi 1. Introduction ............................................................................................................................................... 1 2. Literature Review ...................................................................................................................................... 5 2.1 Clinical Need ........................................................................................................................................ 5 2.1.1 Problems with Current Drug Approval Process ............................................................................ 6 2.1.2 Current Preclinical Two-Dimensional Testing Methods ................................................................ 7 2.1.3 Current Animal Models ................................................................................................................ 9 2.1.4 Cardiovascular 3D Models .......................................................................................................... 10 2.1.5 Cardiotoxicity Testing ................................................................................................................. 12 2.2 Existing Solutions ............................................................................................................................... 14 2.2.1 Live Cell Imaging Microscopes .................................................................................................... 15 2.2.2 Single Well Incubator for Microscope Stages ............................................................................. 16 2.2.3 Multi-Well Incubator for Microscope Stages .............................................................................. 17 2.2.4 Summary of Existing Solutions.................................................................................................... 18 2.3 Need Statement ................................................................................................................................ 19 3. Project Strategy ....................................................................................................................................... 21 3.1 Initial Client Statement ...................................................................................................................... 21 3.2 Stakeholders ...................................................................................................................................... 21 3.3 Initial Objectives ................................................................................................................................ 22 3.4 Constraints ........................................................................................................................................ 23 3.5 Final Objectives ................................................................................................................................. 24 3.6 Revised Client Statement .................................................................................................................. 29 3.7 Project Approach ............................................................................................................................... 29 3.7.1 Management Approach .............................................................................................................. 29 3.7.2 Design Approach ........................................................................................................................ 30 3.7.3 Financial Approach ..................................................................................................................... 30 4. Design Process ......................................................................................................................................... 31 i 4.1 Needs Analysis ................................................................................................................................... 31 4.1.1 Design Needs .............................................................................................................................. 32 4.1.2 Design Wants .............................................................................................................................. 33 4.1.3 Needs and Wants Design Matrix ................................................................................................ 33 4.2 Functions and Specifications ............................................................................................................. 34 4.3 Conceptual Design Phase .................................................................................................................. 37 4.3.1 Brainstormed Design Elements .................................................................................................. 38 4.3.2 Evaluation of Design Elements ................................................................................................... 39 4.3.3 Quantitative Assessment of Design Elements ........................................................................... 53 5. Development and Verification of the Final Design ................................................................................. 56 5.1 Designs with Modified 6-Well Plate Bases ........................................................................................ 56 5.2 Designs with Modified 6-Well Plate Covers Only.............................................................................. 58 5.3 Preliminary Testing of Overall Design Ideas ..................................................................................... 59 5.3.1 Control System ........................................................................................................................... 60 5.3.2 Temperature .............................................................................................................................. 63 5.3.4 Humidity ..................................................................................................................................... 74 5.3.3 Imaging ....................................................................................................................................... 76 5.3.5 CO2 ............................................................................................................................................. 80 5.3.6 Perfusion .................................................................................................................................... 84 5.4 Complete Design of Microscope Incubator System .......................................................................... 88 6. Device Validation and Results ................................................................................................................. 91 6.1 Measuring Temperature ................................................................................................................... 91 6.2 Measuring CO2 Levels ....................................................................................................................... 93 6.3 Measuring Relative Humidity Levels ................................................................................................. 96 6.4 Sterility Testing ................................................................................................................................. 97 6.5 Measuring Flow Rate ........................................................................................................................ 99 6.6 Well Diffusion Test .......................................................................................................................... 100 6.7 Entire Device Testing ..................................................................................................................... 103 6.8 Visualization of Process .................................................................................................................. 105 7. Discussion ...........................................................................................................................................