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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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Customer Need Mapping

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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Customer Need Mapping

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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