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Jay Dolas (BME), Tyler Lisec (ME), Vincent Serianni (BME) RIT

Multidisciplinary Senior DesignProject Readiness Package

Project Title: Automated Microfluidic Cell Separator

Project Number:(assigned by MSD)

Primary Customer:(provide name, phone number, and email)

TBD

Sponsor(s):(provide name, phone number, email, and amount of support)

Preferred Start Term: Fall 2015

Faculty Champion:(provide name and email)

Dr. Blanca Lapizco-Encinas [email protected]

Other Support: Dr. Jennifer Bailey (tentatively)[email protected]

Project Guide:(assigned by MSD)

Jay Dolas06/14/2015 Updated 08/19/2015

Prepared By Date

Received By Date

Items marked with a * are required, and items marked with a † are preferred if available, but we can work with the proposer on these.

RIT – Kate Gleason College of EngineeringMultidisciplinary Senior Design

Project Readiness PackageTemplate Revised Jan 2015


Project Information

The most common technology for sorting cells is flow cytometry, in which laser detection of biomarker tags is used to separate cells of interest. This is an extremely high throughput system that sorts several thousand cells per second and differentiates between up to eighteen cell types. However, it does require the addition of extremely costly fluorescently labeled antibodies, and more than one type may be required to accurately identify a specific cell type. Another method of cell sorting, Magnetic Activated Cell Sorting (MACS), requires the addition of magnetic nanoparticles bound to antibodies. Due to the use of these antibodies, MACS is also an expensive method of cell sorting.

Because of the high cost of these methods, we propose a device which will use microfluidic flow channels applying dielectrophoretics to separate cells based upon their physical and electrical properties. This negates the need for any antibody additions and tagging, greatly reducing the expense associated with cell sorting. These channels will be capable of sorting through as many cell types as the typical flow cytometry instrument. The only consumable material required is the microfluidic flow channel, which must be customized for each cell sorting application.

This system is automated such that the operator will only need to insert the mixed cell type culture into the initial reservoir and start the device. The culture will be drawn through the microfluidic channel and sorted using dielectrophoretics, where the end result will be containers that each hold a single type of cell. Because of the automation, the operator will be able to move on to different tasks as soon as the sorting process has been started. Being a self contained automated unit, this will greatly reduce the chances of human error, contamination, and damaging viable cells while increasing the repeatability, reliability, and time that the operator can spend working on other tasks. Dielectrophoretics is based on the interaction between an electric field and the cells surface and electrical properties, so one standard channel layout can be used to sort a multitude of cells by changing the electric field’s magnitude and frequency.

The potential customer for this automated cell sorting apparatus is anyone who works with a multitude of cells types, cell line development, or simply requires inexpensive high throughput cell sorting. This includes, but is not limited to:

• Medical Laboratories • Medical Device Manufacturers • Research Institutions • Pharmaceutical Testing and Development Companies • Private/Public Biomedical Companies • Regenerative Medicine Companies




Figure 1: Example process of dual frequency dielectrophoretic particle sorting source: http://www.mdpi.com/1422-0067/15/10/18281/htm

Preliminary Customer Requirements (CR): • Partially to fully automated cell separation device (after initialization, cells

separated without further intervention by operator)• Cost effective compared to standard separation methods • Sort cells at reasonable rate • Produces viable cells • Fit and work within a laminar flow hood • Capable of sorting two cell types• Compatible with typical laboratory cleaning solutions • Able to be cleaned and sterilized with existing equipment• Maintains sterility • Ease of use • Ease of repair

Functional Decomposition: 1. Dispense the culture into the initial reservoir 2. Fluid flows into the microchannel 3. Sort each cell type into one of two channels4. Sorted cells flow into final reservoirs




5. Final reservoirs accessible for cell extractionPreliminary Engineering Requirements (ER):

Specification Ideal Value CommentsAutomation Allows user to sort by just adding cells and

starting deviceAccuracy >90% Test using staining techniquesYield >90% Test by counting with hemacytometerViability >90% Test with live/dead stain or trypan blueFlow rate Reynolds Number

< 1Time of operation <10 minsAssembly and Disassembly time

<1 hour

Size 1. X 1.5’ x 10” LxWxH

Constraints: • Fit within a laminar flow hood

o Must be at maximum 1.5’x1.5’x10” (LxWxH) o Weight must be supported by laminar flow hood

• Must be compatible with 120V AC wall outlet • Must be bioinert • Keep the cost of production under $5,000.00 • Device can be easily disassembled for repairs and biology lab standard cleaning

Potential Concepts: 1. Dispense the co-culture into the initial reservoir

a) Automated recovery of co-culture from a flask or petri dish into the initial reservoir

i. Roboticsb) Manually pipette co-culture into the initial reservoir

2. Pump fluid into microchannel a) Micropump b) Vacuum c) Electroosmotic flow d) Mechanical pressure application

3. Sort cells into one of two channels a) Microfluidic spiral flow channel separation

i. Microfluidicsii. Cell culture and analysis

iii. Advanced fluid flow modelingiv. Advanced micro-scale fabrication

b) Dielectrophoretic separation i. Microfluidics

ii. Electronics




iii. Cell culture and analysisiv. Advanced micro-scale fabrication

c) Acoustophoretic separation i. Microfluidics

ii. Electronics iii. Cell culture and analysis iv. Advanced micro-scale fabrication

4. Cell extraction from final reservoir a) Manually pipette from reservoirsb) Detachable reservoirs

i. Fabrication

Project Deliverables: • All design documents (e.g., concepts, analysis, detailed drawings/schematics, BOM,

test results)• Final cost of production estimate • Working prototype • Test plan and results • User manual • Technical paper • Poster • Final presentation materials • Complete Edge site • ImagineRIT presentation/ display

Budget Information: Total Budget Estimate - $3300

Items Projected CostFramework $200Interior Components $3000Disposables $100Software Provided

Intellectual property:• Concern - Silicon Biosystems utilizes dielectrophoretics and microfluidic channels to

separate individual cellso Microarray card for imaging, analysis, and experiments of individual cellso Has a different scope

• Potential to patent as a high-throughput cell sorting device using microfluidics




Project Resources

Required Resources (besides student staffing):

Faculty list individuals and their area of expertise (people who can provide specialized knowledge unique to your project, e.g., faculty you will need to consult for more than a basic technical question during office hours)

Initial/date

Dr. Blanca Lapizco-Encinas: Microfluidics; Dr. Jennifer Bailey: Cell cultureEnvironment (e.g., a specific lab with specialized equipment/facilities, space for very large or oily/greasy projects, space for projects that generate airborne debris or hazardous gases, specific electrical requirements such as 3-phase power)

Initial/date

Biomedical Engineering Student Projects Lab/Wet Lab, MicroE Fabrication LabEquipment (specific computing, test, measurement, or construction equipment that the team will need to borrow, e.g., CMM, SEM, )

Initial/date

Laminar flow hood, incubator, MicroE fabrication equipment, microscope, plate reader, power supply/amplifier (borrowed, purchased, or created)Materials (materials that will be consumed during the course of the project, e.g., test samples from customer, specialized raw material for construction, chemicals that must be purchased and stored)

Initial/date

Aluminum sheeting, electrodes, electrical components, various polymers, cells, culture media, cellular stains

OtherInitial/date

Anticipated Staffing By Discipline:

Dept. # Req. Expected ActivitiesBME 2 Jay Dolas, Vincent Serianni – Cell culture, microfluidicsCEEE 1-2 Power supply creation/modification, amplifier creation/modification,

asymmetric signal splitting and programming for electrode controlISEME 1 Tyler Lisec – Fabrication, fluid flow, mechanicsOther 1 MicroE – Microfluidics/dieletrophoretics fabrication

Skills Checklist:

Mechanical EngineeringME Core Knowledge ME Elective Knowledge

1 3D CAD 3 Finite element analysisMatlab programming Heat transfer

1 Basic machining 1 Modeling of electromechanical & fluid systems




ME Core Knowledge ME Elective Knowledge3 2D stress analysis Fatigue and static failure criteria3 2D static/dynamic analysis 3 Machine elements2 Thermodynamics Aerodynamics1 Fluid dynamics (CV) 3 Computational fluid dynamics

LabView 1 Biomaterials2 Statistics Vibrations1 Materials selection IC Engines

GD&TLinear ControlsCompositesRoboticsOther (specify)

Electrical EngineeringEE Core Knowledge EE Elective Knowledge

1 Circuit Design (AC/DC converters, regulators, amplifies, analog filter design, FPGA logic design, sensor bias/support circuitry)

Digital filter design and implementation

1 Power systems: selection, analysis, power budget Digital signal processingSystem analysis: frequency analysis (Fourier, Laplace), stability, PID controllers, modulation schemes, VCO’s & mixers, ADC selection

Microcontroller selection/application

1 Circuit build, test, debug (scope, DMM, function generator

Wireless: communication protocol, component selection

Board layout Antenna selection (simple design)Matlab Communication system front end designPSpice Algorithm design/simulationProgramming: C, Assembly Embedded software design/implementation

1 Electromagnetics: shielding, interference Other (specify)

Industrial & Systems Engineering n/a

Biomedical EngineeringBME Core Knowledge BME Elective Knowledge

3 Matlab Medical image processing1 Aseptic lab techniques COMSOL software modeling3 Gel electrophoresis Medical visualization software

Linear signal analysis and processing 2 Biomaterial testing/evaluation1 Fluid mechanics Tissue culture1 Biomaterials 1 Advanced microscopy3 Labview 1 Microfluidic device fabrication and measurement

Simulation (Simulink) Other (specify)System physiologyBiosystems process analysis (mass, energy balance)

3 Cell cultureComputer-based data acquisition

2 Probability & statisticsNumerical & statistical analysis




BME Core Knowledge BME Elective KnowledgeBiomechanics

Computer Engineering n/a



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