Oliveri My Little Bioreactor Serrano, Tomas Blaquier ...
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My Little BioreactorGroup 11: Alessandro Leggio, Ashley Cador, Ava Nemanic, Gabriel Caballero, Rodrigo Moreno, Sergio Serrano, Tomas Blaquier, Tyler Oliveri
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My Little Bioreactor
Dynamic Control
Fluid Handling
Gas Management
Waste Management
Optical Sensing
Thermal Control
Translation Control
Enclosure
Prototype Cost: $7681.55
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● Reliability○ Reducing component failure modes
● Manufacturability○ In-house production capabilities
● Reduced Cost○ Modular design with full bioreactor
capabilities
Motivations for Design
Reduced Costs
Reliability
Manufacturing
Simplicity
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Enclosure
1800W Power SupplyTwist Emergency Stop Button
40-Pin 7” LCD Display
Raised Indicator Light
Hatch Door
55cm50cm
50cm
● 304 Stainless Steel Plates○ Limits outside heat transfer
● Aluminum Frame Bars○ Fastened with M3 screws
●
● Hatch Doors○ 45cm opening○ Friction Hinges ○ Push-Lock to secure
●
Prototype Cost: $525.65
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Heat Transfer - Wall Plate Wall Section View
Inside Outside
Ti = 70°C
To = 40°C
Heat Transfer
Kss - 304 Stainless Steel Conductivity
Lss,nom - Nominal Wall Thickness (1.5mm)
To,1 - Max External Temp (40°C)
Lss,d - Desired Wall Thickness
To,2- Max External Temp + Buffer (50°C)
Ld,ss = 2.25mm
12 Gauge 304 Stainless Steel (2.66mm)
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Customer Need Mapping
Enclosure
2. Cost < 10,000$
14. Exterior surface not too hot to touch
9. Includes visual indicator light
8. Includes user interface
7. Includes emergency
shut-off6. Easy interior
access
5. Runs from 120 VAC outlet
4. Fits on research benchtop
3. Moveable by one person after disassembly
4.8 mm wallthickness
Light 2” from LCD
7 user commands 3 second kill
time45 cm
opening
64.01 Watts
3,150 cm2
footprint
38.3kg weight$516.22
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Thermal Control
OTS refrigeration pump (455W)
Infrared Non-Contact Sensor
Circulating liquid cooling/heating system
Conduction heat
exchanger
Resistive heating element (2500W)
Prototype Cost: $525.65
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Thermal ControlCooling heat exchangerHeating Element
Dehumidifier
Water Pump
● Rated power = 16.8 W● Pump head = 7m● Mass flow rate = 10L/m
● 17.1g water storage
Controlled heat generation and removal
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Thermal ControlCopper Sheet
Contact Temp Sensor
Infrared Sensor
Gasket
140 mm 182 mm
Avg. accuracy: ±0.55°C
ABS milled stock
Water line fittings
TEST TUBE RACKWELL PLATE
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Thermal ControlDifferent channel design leads to different velocity distribution, which results in a difference in temperature distribution.
Heat Transfer rate → ABS Heat Transfer rate → Copper
q
q
copper
ABS
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Thermal Control
Time (s) Time (s)
Tem
per
atu
re (°
C)
Water reservoir Test tube
Temperature gradient analysis
Reservoir target temp = 80°C
15mL test tube requires 539.7 J of energy to reach maximum setpoint
MATLAB nodal analysis with varying heat transfer rate for both reservoir and test tube temperatures
Setpoint temp (70°C) reached in 10.53 min
Q
Tem
per
atu
re (°
C)
Step time 1 sec
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Thermal
2. Cost < 10,000$
1. Lifetime > 10 years
24. Reach setpoint temp <
15 min
23. Humidity removal
22. Uniform heating/cooling 21. Internal temp
at ± 2.5°C
20. Operating temp range
19. Incubation period > 2 weeks
5. Runs from 120 VAC outlet
15 years
10.53 min
17.1g10.53 min +
Shaking ± 0.55°C
0°C - 100°C
15 years
455 W$525.65
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Translation Control
Prototype cost: $416.34YX
Z
LDO 17H Stepper Motors
M10x2mm Carbon Steel Lead Screws
6061 Aluminum Mounts & Carriages
Multi-Purpose Tool Head
Manganese Bronze Nuts
Stainless Steel Guiding Rod
Timing Belt Pulley & Tensioner
Lead Screw Gantry System
24cm
24cm
8cm
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Translation Control
Mass = 1956.14g
Where;
42STH40-1004MAH motor torque = 0.3825 Nm
⍺
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Translation Control
Mass = 1063.48g
M10x2mm Lead screw deflection
F = 10.43 N
L = 300mm
D = 10mmLinear velocity
Total system power consumption = 17.6 W
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Translation
2. Cost < 10,000$
33. Linear velocity 20. Operating temp range
18. Coverage range
5. Runs from 120 VAC outlet
5.625 mm/s
0-90°C
0.073m
17.6 W$409.07
1. Lifetime > 10 years
15 years
2
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Optical Sensing:Optical Density
Pico LEDs3mm LEDs
● Capable of OD measurements in conical tubes and standard well plates
● LabJack T7 DAQ with 86 Analog and 23 Digital Inputs controlled by RaspberryPi computer
LED Scale:
140 mm
182 mm
120 mm
Prototype cost: $2912.65
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Optical Sensing:OD/FI Tool
Identical surfaces for Well Plate and Tube Measurements
Central ALS Strictly for OD600
4 sets of filtered ALS and LED for up to 4 possible FI measurements
● Highly customizable for various desired wavelength measurements
● Easy to disassemble and replace
Filtered LEDs provide light for FI measurements
Row of 5 Specifically filtered ALS take OD&FI measurements
Direct mounting to Gantry System multi-purpose tool head
53 mm
19 mm
29 mm
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Optical Sensing:Fluorescent Intensity
Color Wavelengths (nm) Detectable Fluorophores
Blue 407, 430, 460 BOBO-1, DAPI, Marina Blue, NBD-X, mTFP1, SpectrumAqua, SpectrumBlue, TagBFP, ThiolTracker Violet GSH
Green 540, 555 tdTomato, mRFP, 5-carboxynapthofluorescein, 6-carboxyrhodamine 6G
Purple 360 Marina Blue, Vybrant DyeCycle Violet
Red 600, 630 mKate2, Nile red-phospholipid, SNARF-1, TurboFP635, YOYO-3
Yellow 573 LysoTracker Red, Rhodamine Red-X, TurboRFP
Source: ThorLabs
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Optical Sensing
2. Cost < 10,000$
1. Operating Lifetime > 10 Years
33. < 6.5 min processing time
for 384 well plate
32. Non-lethal wavelengths
31. Measures fluorescent
intensity30. Measures optical density
27. Includes a photobioreactor
mode
10. Is programmable
5. Runs from 120 VAC outlet
4.8 mm wallthickness
< 25 ms sensor
settling time
All lights filtered above
310 nm9 filters for 20+ fluorophores
600 nm filters with ambient light sensors
and LEDs
LED strips with 450 nm
wavelength
3 different mechanisms
29.1 W$2912.65
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Dual Axis Guide Rails
Dynamic Control
Adjustable Kinematic Linkages
15.5 cm
27 cm
Shaker Plate
Nema 17 H-Class Stepper Motors
Prototype Cost: $723.94
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Dynamic Control Shaking Characteristics
Orbital shaking minum ranges
● Orbit Diameter: 25 - 50 mm
● Frequency: 50 - 300 rpmCell De-clumping
Nutrient Distribution
Oxygen Aeration
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Dynamic Control Kinematic Linkages
Linkage Length
PackagingPattern Shape Pattern Size
*Pictured Linkage Simulation: 50 mm orbital diameter and corresponding double orbital pattern
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Dynamic Control AdjustabilityDesign Iteration 1.0 Design Iteration 2.0
Continuously adjustable primary & secondary linkages
No disassembly required
Continuously adjustable secondary linkage
Primary Linkage:● adjustable in 0.4 mm increments● Requires disassembly
Easy Access Knob
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Dynamic Control Adjustability
Female rod end
Adjustable Knob● Tick marks every 0.2mm
Set screw access
Custom lead screw
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Dynamic Control AccessibilityUser’s view while adjusting primary linkages:
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Dynamic Control AccessibilityUser’s view while adjusting secondary linkages:
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Dynamic Control
2. Cost < 10,000$
1. Lifetime > 10 years
29. Independent
shaking patterns
28. Shakes well plates and tubes in Linear, Orbital, and Double
Orbital patterns
26. Accommodates conical tubes of
sizes: 15mL & 50mL
25. Accommodates well plates of the
sizes: 6, 24, 48, 96, deep 96, 384
5. Runs from 120 VAC outlet
10+ years
N/A
3 patterns
2 sizes
6 sizes
8.2 W$723.94
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Fluid HandlingStepper Motorized MiNi SY-04 Syringe Pump
Multiport Selector Valve
500mL Beaker Reservoir
L16-P Miniature Linear Actuator
PPS Plastic Y-Connector
Custom PP Pipette
Prototype Cost: $1,463.00
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Distal Dispensing L16-P Miniature Linear Actuator
PPS Plastic Y-Connector
Custom PP Pipette
2024 Aluminum Mount PlateReynold’s Number
Q - Desired Flow Rate (225 μL/s to 300 μL/s)D- Pipette Tip Diameter (0.38mm)
→ Laminar Flow: No Aerosols
Linear Actuator:
● Actuator Stroke : 100mm● Maximum Force: 50N (11lbs)
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Pump and Valve SystemHagen-Poiseuille Equation
L- Length of Pipette (0.125m)Q - Desired Flow Rate (225 μL/s to 300 μL/s)A- Cross Sectional Area of Pipette Tip
Syringe Pump:
● Volume: 20 ml● Liquid Pressure: 0-1.2 MPa● Accuracy Resolution: ±0.4µL
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Fluid Handling
11. Only nonporous materials contact
cell cultures
40. No cross contamination
39. Dispensing volume precision
±0.01 μL 38. Dispensing
volume accuracy of ±0.1μL
37. Aliquot fluid volume from 5–20,000 μL
36. Achieves dispense rates from 225 μL/s to 300 μL/s
35. Dispenses fluid without creating
aerosols
34. Is capable of automated liquid
handling
0.45 Diameter
Ratio
±0.4µL20 ml Aliquot
Volume
ΔP = 0.065MPa
Reynolds Number = 1126
2 degrees of automation
30 µm pore size
±0.4µL
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Liquid Waste
● Fluid handling valve and pump are
plumbed into both sterilization
fluids and the waste container
Waste Management
1 Gallon Autoclavable
Polypropylene Waste Container
Glass sterilization fluid flasks
500 mLFormaldehyde Solution
1000 mL70% Isopropyl
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Waste Container
1
Waste Management Sterilization Operation
Well Plate or Tube
Sterilization Reservoir(s)
2
Liquid Waste
Sterilization Fluid
4Contaminated
Sterilization
Fluid
Pipette tip submerges
itself in sterilization
fluid
3
Air/N2/He purge fluid lines evaporate residual Isopropyl5Sterilization Time
for typical operation:
4-30 minutesWaste container and
contents are autoclaved6
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Fan and vent hatches are opened
Fan pulls air into enclosure
Air escapes from top vent
into biosafety cabinet
Waste Management Gaseous Waste
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Customer Need Mapping
Waste Management
2. Cost < 10,000$
1. Operating Lifetime > 10 Years
15. Sequester and Neutralize
own waste13. BSL-2 Space
Operation
11. Only Non-porous
materials
5. Runs from 120 VAC outlet
10000 hours at
continuous use
Formaldehyde & Isopropyl via Fluid handling piette -No leakage
-Decontamination-Cleaning-Venting
Material pores << 4um
38 Watts$186.65
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Gas Management
Solenoid Valves● 12 V● 20 W
Gas Manifold● Max. <1000 PSI
Pressure Transducer● 4.5 V● <100 PSI
Mass Flow Meter● Min. 0-20 sccm● Max. 0-500 L/min
Pressure Relief Valve● Max temp 82°C● <300 PSI
Prototype cost: $944.39
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Gas ManagementPipette injection sitePressure Relief Silicone valves for test tubes:
● Assists in maintaining
independent environmental
conditions (Req. 17)
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Gas ManagementVolumetric Flow Rate
Continuity Equation
Bernoulli’s Equation
Pressure difference due to change in tubing diameter was determined to be minimal (0.2 Pa)
12.7 mm 4 mm
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Gas
1. Lifetime of at least 10
years18. Inject, measure, regulate N2, O2, CO2, CH4, H2
17. Maintains independent
environmental conditions
16. Cultures microbes in
closed vessels12. Non-reactive
materials
10. Programmable
5. Runs from 120 VAC outlet
2. Cost < 10,000$
6 solenoid valves with mass flow
meter
5 independent conditions
4 vessel types 0
compounds
1 sensor
64.01 Watts
$944.39
10 years
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