automated microfluidic cell separator project group: 16083
TRANSCRIPT
Automated Microfluidic Cell Separator
Project Group: 16083
The P16083 Group
Member Major Role Contact
Jay Dolas BME Lead Engineer [email protected]
Alexandra la Londe BME Microfludics Specialist [email protected]
Vincent Serianni II BME Project Manager [email protected]
Tyler Lisec ME Lead Mechanical [email protected]
Ryan Kinney EE Lead Electrical [email protected]
Chris Molinari EE Controls Engineer [email protected]
“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
Background Summary – What is Microfluidics?
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
http://cnx.org/resources/7b4d03a7fc1e724f75950258ae6d2356
Background Summary - What Size of Particles?
Research and Testing Teaching Tool General Medical
Purposes
http://www.mdpi.com/1422-0067/15/10/18281/htm
Background Summary – Why Sort Cells?
http://eshop.eppendorf.ca/upload/productView/Eppendorf_5427R_high-capacity_centrifuge.jpg
Background Summary - Other Sorting Methods
Centrifuge Flow Cytometry Channel Geometry Magnetic Based
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.
Project Statement
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
Hydrodynamic Cell Separator Inertial forces to separate cell types
High shear forces can damage cells
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
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
Project Goals and Deliverables
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
Key Constraints
Use Scenarios – Teaching Aid
Use Scenarios – Medical Field
http://www.rheonix.com/corporate/careers.phphttps://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
Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important
Customer Requirements
Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important
Customer Requirements
Separation can be visualized
Engineering Requirements
Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important
Engineering Requirements
Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important
n = Less Importantn = Moderate Importancen = Most Important
House of Quality
Project Plans
MSD I: 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
Project Plans (First 3 Weeks)
Project Plans (Next 3 Weeks)
Project Plans (MSD I)
Risks
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Technical
Power Supply Failure
Size/Weight
Channel Fabrication Errors
Fluid Flow Pressure
Cells Having Similar Electrical Properties
Cell Viability
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Resource
Expense of Components
Lack of Manpower
Data Gathering
Cells for sorting
Clean Room Workers
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
Safety
Electric Shock
Lab Work
Channel Fabrication
Leaks
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Environmental & Societal
Waste Generation
Potential Air Born Chemicals
Disposal of Chemicals
Class Room Setting