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

Multidisciplinary Senior Design

Project 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

Other Support: / Dr. Jennifer Bailey (tentatively)

Project Guide:
(assigned by MSD)
Jay Dolas / 06/14/2015 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.

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 channels

4. Sorted cells flow into final reservoirs

5. Final reservoirs accessible for cell extraction

Preliminary Engineering Requirements (ER):

Specification / Ideal Value / Comments
Automation / Allows user to sort by just adding cells and starting device
Accuracy / >90% / Test using staining techniques
Yield / >90% / Test by counting with hemacytometer
Viability / >90% / Test with live/dead stain or trypan blue
Flow rate / Reynolds Number < 1
Time of operation / <10 mins
Assembly 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:

Dispense the co-culture into the initial reservoir

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

Robotics

b) Manually pipette co-culture into the initial reservoir

Pump fluid into microchannel

a) Micropump

b) Vacuum

c) Electroosmotic flow

d) Mechanical pressure application

Sort cells into one of two channels

a) Microfluidic spiral flow channel separation

Microfluidics
Cell culture and analysis
Advanced fluid flow modeling
Advanced micro-scale fabrication

b) Dielectrophoretic separation

Microfluidics
Electronics
Cell culture and analysis
Advanced micro-scale fabrication

c) Acoustophoretic separation

Microfluidics
Electronics
Cell culture and analysis
Advanced micro-scale fabrication
Cell extraction from final reservoir

a) Manually pipette from reservoirs

b) Detachable reservoirs

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 Cost
Framework / $200
Interior Components / $3000
Disposables / $100
Software / Provided

Intellectual property:

• Concern - Silicon Biosystems utilizes dielectrophoretics and microfluidic channels to separate individual cells

o Microarray card for imaging, analysis, and experiments of individual cells

o 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 culture
Environment (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 Lab
Equipment (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
Other / Initial/date

Anticipated Staffing By Discipline:

Dept. / # Req. / Expected Activities
BME / 2 / Jay Dolas, Vincent Serianni – Cell culture, microfluidics
CE
EE / 1-2 / Power supply creation/modification, amplifier creation/modification, asymmetric signal splitting and programming for electrode control
ISE
ME / 1 / Tyler Lisec – Fabrication, fluid flow, mechanics
Other / 1 / MicroE – Microfluidics/dieletrophoretics fabrication

Skills Checklist:

Mechanical Engineering

/ ME Core Knowledge / ME Elective Knowledge /
1 / 3D CAD / 3 / Finite element analysis
Matlab programming / Heat transfer
1 / Basic machining / 1 / Modeling of electromechanical & fluid systems
3 / 2D stress analysis / Fatigue and static failure criteria
3 / 2D static/dynamic analysis / 3 / Machine elements
2 / Thermodynamics / Aerodynamics
1 / Fluid dynamics (CV) / 3 / Computational fluid dynamics
LabView / 1 / Biomaterials
2 / Statistics / Vibrations
1 / Materials selection / IC Engines
GD&T
Linear Controls
Composites
Robotics
Other (specify)

Electrical Engineering

/ EE 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 processing
System 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 design
PSpice / Algorithm design/simulation
Programming: C, Assembly / Embedded software design/implementation
1 / Electromagnetics: shielding, interference / Other (specify)

Industrial & Systems Engineering

n/a

Biomedical Engineering

/ BME Core Knowledge / BME Elective Knowledge /
3 / Matlab / Medical image processing
1 / Aseptic lab techniques / COMSOL software modeling
3 / Gel electrophoresis / Medical visualization software
Linear signal analysis and processing / 2 / Biomaterial testing/evaluation
1 / Fluid mechanics / Tissue culture
1 / Biomaterials / 1 / Advanced microscopy
3 / Labview / 1 / Microfluidic device fabrication and measurement
Simulation (Simulink) / Other (specify)
System physiology
Biosystems process analysis (mass, energy balance)
3 / Cell culture
Computer-based data acquisition
2 / Probability & statistics
Numerical & statistical analysis
Biomechanics

Computer Engineering

n/a

RIT – Kate Gleason College of Engineering
Multidisciplinary Senior Design / Project Readiness Package
Template Revised Jan 2015