1. Automated Microfluidic Cell Separator Project Group: 16083

2. Table of Contents  Group Members  Background Summary  Project Statement  Use Scenarios  Stakeholders  Customer Requirements  Engineering Requirements  House of Quality  Project Plan  Risk Management

3. The P16083 Group MemberMajorRoleContact Jay DolasBMELead Engineerjrd8174@rit.edu Alexandra LaLondeBMEMicrofludics Specialistael9930@rit.edu Vincent Serianni IIBMEProject Managervcs7133@rit.edu Tyler LisecMELead Mechanicaltjl9229@rit.edu Ryan KinneyEELead Electricalrhk9387@rit.edu Chris MolinariEEControls Engineercxm4025@rit.edu

4.  “Microfludics is the field that studies the manipulation of small amounts of fluids (10 -9 to 10 -18 liters)”  “Microfluidics refers to the handling of liquids or gases at a scale generally below 1mm, where a number of phenomena that are NOT present or not predominant at larger scales can be exploited for numerous purposes”.  “The field of microfluidics is in essence multidisciplinary as it combines microfabrication techniques with chemistry and biology”. BIME 489-01 Dr. Blanca Lapizco-Encinas Background Summary What is Microfluidics?

5. Background Summary What is Dielectrophoretics?  Dielectrophoresis is the movement of particles due to polarization effects in a non-uniform electric field.  In the presence on a nonuniform electric field, one side of the dipole will be in a region with a lower field intensity.  This will produce and UNEVEN charge alignment in the particle, inducing it to move toward the regions of greater field strength BIME 489-01 Dr. Blanca Lapizco-Encinas

6. http://cnx.org/resources/7b4d03a7fc1e724f75950258ae6d2356 Background Summary What Size of Particles?

7.  Research and Testing  Teaching Tool  General Medical Purposes http://www.mdpi.com/1422-0067/15/10/18281/htm Background Summary Why Sort Cells?

8. http://eshop.eppendorf.ca/upload/productView/Eppendorf_ 5427R_high-capacity_centrifuge.jpg Background Summary Other Sorting Methods  Centrifugation  Flow Cytometry  Channel Geometry  Magnetic Based

9. A cell separator is a device that separates cells in a mixture, based upon pre-established criteria (biomarkers, size, electrical characteristics, etc.). This is necessary in many cell culture and diagnostic applications where downstream processes occur after cell culture, such as purification or analysis. Optimally, this device should not interfere with the viability or characteristics of the cells, while still being cost effective. Current cell separation devices require some sort of labeling (either fluorescent or magnetic) which is not only costly but can affect cell behavior and mortality. We propose an automated microfluidic system that utilizes developing technologies (dielectrophoretics) to reduce costs drastically while maintaining cell viability. The goals of this project are to develop a system that not only sorts cells without the use of labeling, but also fits within a biosafety cabinet, is self-driven, and is automated (hands- off once the sample is loaded and sequence has started). The expected result is a functional prototype that fits all of the goals above and is suitable for use in a teaching laboratory. The design and prototype must conform to intellectual property and diagnostic laboratory standards so that it may be marketed this as a definitive step forwards in cell separation technology. Problem Statement

10. http://www.appliedcytometry.com/flow_cytometry.php http://www.elveflow.com/microfluidic-tutorials/cell-biology-imaging-reviews-and- tutorials/microfluidic-for-cell-biology/label-free-microfluidic-cell-separation-and- sorting-techniques-a-review/ Current State  Flow Cytometry  Fluorescence labeling  Laser to excite and identify the cells  Additives could alter or damage cells  Labeling is expensive  Hydrodynamic Cell Separator  Inertial forces to separate cell types  High shear forces can damage cells  Channel design and manufacturing is timely and expensive

11.  Dielectrophoresis  Uses electric fields to manipulate the cell location in a stream  No need for additives  No added shear force http://www.elveflow.com/microfluidic-tutorials/cell-biology-imaging- reviews-and-tutorials/microfluidic-for-cell-biology/label-free- microfluidic-cell-separation-and-sorting-techniques-a-review/ Desired State

12. Working prototype that can:  Demonstrate in a class setting the use of dielectrophoretics in cell separation  Act as a partially automated system  User only has to load the sample and set the target specifications  Maintain cell viability during sorting process  Accurately sorts the target cells Documentation of the prototype that illustrates:  Proper use and care of the device  Target specifications  Voltage amplitude and frequency standards to sort a given cell type Project Goals and Deliverables

13.  Device start-up cost below $5000  Lightweight - able to be moved by 1 person  Electrically shielded and insulated  Bio-hazard containment  Footprint (1.5' x 1.5')  120V outlet compatible  Reusable channel  Perform process within one hour Key Constraints

14. Use Scenarios – Teaching Aid

15. Use Scenarios – Medical Field

16. http://www.rheonix.com/corporate/careers.php https://twitter.com/ritbme Stakeholders  Customer – Dr. Blanca Lapizco-Encinas  End Users – Lab Workers, Professors, Researchers, Students  Potential Sponsors – Rheonix Inc. or the BME Department at RIT  Other Stakeholders – P16083 Group, MSD Team

17. Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important Customer Requirements

18. Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important Customer Requirements Separation can be visualized

19. Engineering Requirements Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important

20. Engineering Requirements Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important

21. = Less Important = Moderate Importance = Most Important House of Quality

22. Project Plans MSD I:  Define problem and requirements  Interact with the Customer  Interview, Funding, Customer vs Engineering Requirements  Gather data for design and hardware  Research, Design, Review, Standards, Bill of Materials (BOM)  Validate channel designs and characterize cell behavior MSD II:  Build validated designs  Circuits, PDMS Channels, 3D Print (where needed), Subsystems  Benchmark device and compare to engineering requirements  Present project in working state  Imagine RIT, Class Demo

23. Project Plans (MSD I)

24. Project Plans (First 3 Weeks)

25. Project Plans (Next 3 Weeks)

26. Risks

27. http://www.micromanufacturing.com/sites/default/files/stories/ import/1002-Fluids-web-images/micronitimage_opt.jpeg Technical  Power Supply Failure  Size/Weight  Channel Fabrication Errors  Fluid Flow Pressure  Cells Having Similar Electrical Properties  Cell Viability

28. http://images.clipartpanda.com/no-money-sign-clip-art-money-sign.png Resource  Expense of Components  Lack of Manpower  Data Gathering  Cells for sorting  Clean Room Workers

29. http://www.health-safety- signs.uk.com/productimage s/Danger-Electric-shock- risk-sign.gif http://www.emedco.com/media/catalog /product/Indl-Eyewash-Shower--First- Aid-Signs-42878BBVPLY2WY-ba.jpg Safety  Electric Shock  Lab Work  Channel Fabrication  Leaks

30. http://cdn2.bigcommerce.com/server4600/10c6f/ products/1621/images/3022/WS32001_warning _airborne_toxic_chemicals__69205.1403023543.1280.1280.png?c=2 Environmental & Societal  Waste Generation  Potential Airborne Chemicals  Disposal of Chemicals  Teaching Laboratory Setting

31. Questions and Comments