Automated Microfluidic Cell Separator Project Group: 16083 1

The P16083 Group Member Major Role Contact Jay Dolas BME Lead Engineer jrd8174@rit.edu Alexandra la Londe Microfludics Specialist ael9930@rit.edu Vincent Serianni II Project Manager vcs7133@rit.edu Tyler Lisec ME Lead Mechanical tjl9229@rit.edu Ryan Kinney EE Lead Electrical rhk9387@rit.edu Chris Molinari Controls Engineer cxm4025@rit.edu 2

Background Summary – What is Microfluidics? “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 tecniques with chemistry and biology”. BIME 489-01 Dr. Blanca Lapizco-Encinas 3

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 4

Background Summary - What Size of Particles? http://cnx.org/resources/7b4d03a7fc1e724f75950258ae6d2356 5

Background Summary – Why Sort Cells? Research and Testing Teaching Tool General Medical Purposes http://www.mdpi.com/1422-0067/15/10/18281/htm 6

Background Summary - Other Sorting Methods Centrifuge Flow Cytometry Channel Geometry Magnetic Based http://eshop.eppendorf.ca/upload/productView/Eppendorf_5427R_high-capacity_centrifuge.jpg 7

Project Statement 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.

Current State Flow Cytometry Hydrodynamic Cell Separator Fluorescence labeling Laser to excite and identify the cells Additives could alter or damage cells Hydrodynamic Cell Separator Inertial forces to separate cell types High shear forces can damage cells 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/

Desired State 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/

Project Goals and Deliverables Working prototype that can: Separate cells to 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 for certain cells Voltage amplitude and frequency standards in order to sort a given cell

Key Constraints Device start-up cost Weight, to be moved by 1 person Electrical shielding and insulation Bio-hazard containment Footprint (2' x 1') 120V outlet compatible Reusable channel Channel is visible under microscope during run Preform process within one hour

Use Scenarios – Teaching Aid 13

Use Scenarios – Medical Field 14

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 http://www.rheonix.com/corporate/careers.php https://twitter.com/ritbme 15

Customer Requirements Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important 16

Customer Requirements Separation can be visualized Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important 17

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

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

House of Quality n = Less Important n = Moderate Importance n = Most Important

Project Plans MSD I: MSD II: Introduced to the project and group Gather data for design and hardware Research, Design, Review, Standards, Bill of Materials (BOM) Interact with the Customer Interview, Funding, Customer vs Engineering Requirements MSD II: Build validated designs Circuits, PDMS Channels, 3D Print (where needed), Subsystems, Benchmarking Present project in working state Imagine RIT, Class Demo 21

Project Plans (First 3 Weeks) 22

Project Plans (Next 3 Weeks) 23

Project Plans (MSD I) 24

Risks 25

Technical Power Supply Failure Size/Weight Channel Fabrication Errors Fluid Flow Pressure Cells Having Similar Electrical Properties Cell Viability http://www.clker.com/cliparts/O/c/x/E/f/v/fire-hazard-md.png 26

Resource Expense of Components Lack of Manpower Data Gathering Cells for sorting Clean Room Workers http://images.clipartpanda.com/no-money-sign-clip-art-money-sign.png 27

Safety Electric Shock Lab Work Channel Fabrication Leaks 28 http://www.health-safety-signs.uk.com/productimages/Danger-Electric-shock-risk-sign.gif http://www.emedco.com/media/catalog/product/Indl-Eyewash-Shower--First-Aid-Signs-42878BBVPLY2WY-ba.jpg 28

Environmental & Societal Waste Generation Potential Air Born Chemicals Disposal of Chemicals Class Room Setting http://cdn2.bigcommerce.com/server4600/10c6f/ products/1621/images/3022/WS32001_warning _airborne_toxic_chemicals__69205.1403023543 .1280.1280.png?c=2 29