bacteria actuation, sensing, and transport (bast) in micro/nanoscale
DESCRIPTION
Bacteria Actuation, Sensing, and Transport (BAST) in Micro/Nanoscale. Dr. MinJun Kim Dept. of Mechanical Engineering & Mechanics Drexel University. Layout of This Presentation. 1. Introduction of Flagellated Bacteria. - PowerPoint PPT PresentationTRANSCRIPT
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Bacteria Actuation, Sensing, and Transport (BAST) in
Micro/Nanoscale
Dr. MinJun Kim
Dept. of Mechanical Engineering & Mechanics
Drexel University
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Layout of This Presentation
2. Microscale Bacterial Actuation - Chaotic Microfluidic Mixing System- Chemotactic Bacterial Sensing System- Self-sustained Microfluidic Pump- Autonomous Bacterial Transportation System
4. Microbial Risk Assessment
- Ultra-fast Bacteria Detection and Configuration- Rapid Bacteria Cell Lysis
1. Introduction of Flagellated Bacteria
5. Conclusions & Acknowledgements
3. Nanoscale Bacterial Actuation - Nanoscale Mechanical Actuator- Flagella-templated Nantube
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Going Micro & Nano: Miniaturization Theory
Why do we need it?
- Reduced fabrication cost
- Reduced sample consumption
- High sample throughput
- Superior performance (speed / efficiency)
- MEMS and NEMS compatible
What are the applications?
- Molecular separations
- Chemical and biological synthesis
- Medical and clinical diagnostics
- Environmental monitoring
- DNA sequence analysis
- Process control
Why not use “nature”?
- Challenge to micron-nano scale actuation
- Intergration Engineering with Biology
Self-powered Bacterial Pump
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Flagellated Bacteria (E.coli & Serratia marcescens)
Flagellated Bacteria:- Cell body & Flagella- Rod-shaped cell body : 2 m long, 1 m diameter- Flagella : rotary motor, hook, and filament
10 m
2 m
1m
A cell of E. coli, fluorescently labeled.(Turner, Ryu, and Berg 2000)
http://www.npn.jst.go.jp/ Namba
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E.coli Rotary Motor, Hook, and Filament
25 nm
Filament – typically about 10 m and 20 nm in diameter. Helical shape in the unstressed state.
Hook – about 50 nm long and 20 nm in diameter. Plays the universal joint.
Motor – proton (H+) is the energy source. The typical rotation speed is about 100 Hz. The motor can rotate either direction.
Schematic diagram (Berg, 2003), electron micro-scopy image of the flagella motor (De Rosier, 1998), and http://www.npn.jst.go.jp/Namba
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E.coli in Motion
Sequence of E. coli flagella bundling(Turner, Ryu, and Berg, 2000)
E. coli swim by rotating helical filaments. Filaments form a bundle and disperse the
bundle. Tumbles and runs change the swimming
directions.
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Macro-scale Model of Bacterial Flagellar Bundling
Model Full-Scale
Fluid 10 5 cP 1cP
Flagella: 10 cm 10 um
Rotation: 0.3 Hz100 Hz
SetupTwo stepper motors.Epoxy-filled plastic tubes in helical shape.High viscosity silicone oil (100,000 cp).
Match geometryPitch, Aspect ratio, Number of turns.
Match flow characteristicsReynolds number 10-3 (Re 10-5 for Bacteria).
[ FRONT VIEW ] [ SIDE VIEW ]
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Flow by Bundles Helices
- Flexible Helices Movie (Real time)
- Complex flow field induced by bundling
- Bundled state resembles single helix
flow
~
Bundled helices Double thickness helix
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Test Geometry & Experimental Setup
y
x24 mm 20 mm 16 mm 12 mm 8 mm 4 mm 0.5 mm
Fluorescence
No Fluorescence
Width = 200 m
Depth = 40 m
(a)
(b)
• PDMS Microchannels Using Soft-Lithography Techniques
• E.coli: Tumbly (RP 1616), Wild type (HCB 33), and Immobile
• Concentrations of E. Coli: 0 ~ 109 /ml
• Flowrate: 0.5 ~ 1.25 l/min
• Velocity: 1 ~ 2 mm/s
(a) Buffer + 0.02% of FITC + Dextran (MW=77,000) 0.97 cp @ 24.3 C
(b) Buffer + 0.02% of Dextran (MW=68,800) 0.98 cp @ 24.3 C
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Bacteria-Enhanced Diffusion
<Baseline> <Immobile E.coli>
<Tumbly E.coli> <Wild type E.coli>
Fixed at
U = 1.04 mm/s
x = 24 mm.
Each Concentration of E.coli = 1.05 109/ml. channel wall
Flow
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Chemotactic Bacteria-Sensors
1
7
6
5
4
3
2
y
x
1.
3.
5.
7.
1
2
3
Bacteria’s Chemotatic Receptors
Sudden Change
Rotary Motor Performance Affected
Global Microfluidic Effects
Monitoring or Detecting
Bio-Sensor
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Controlled-Mixing in Microfluidics
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Formation of Bacterial Carpets
• Concentration of Serratia Marcescens
: 2 ~ 5 109/ml
• Time : ~ 1 hour
• Flow rate : 0.06 l/min
On : 10 seconds
Off : 5 minutes repeatedly
Flow
…etc…
…etc…
Glass wall
PDMS wall
15 m
Fill
fa
cto
r [%
]
Time [s]10
010
110
210
310
410
0
101
102
PDMSGlass
17 6 5 4 3 2
y
x
2
1
3
1
23
MJ Kim and KS Breuer, PNAS, 2007
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Cell Orientation on Bacterial Carpets
-100 -50 0 50 1000
0.002
0.004
0.006
0.008
0.01
0.012
PD
FCell Orientation [Degree]
-30 < degree < 30 : 54.9013%
-40 < degree < 40 : 64.3485%
-50 < degree < 50 : 74.5086%
Single Cell : 80.5192 %
Group Cell : 19.4808 %
Bacterial Carpet: 50 m x 50 m
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Chaotic Mixing with Bacterial Carpet
Baseline Dead Bacterial Carpet Live Bacterial Carpet
Depth: 15 mWidth: 200 m
0 5 10 15 20 250
2
4
6
8
10x 10- 7
D [
cm
2/s
]
Concentration of Bacteria [ x 108/ml]
BaselineDeadTumblyWild- type (1)Wild- type (2)Wild- type (3)
Baseline
Dead Bacterial Carpet
Live Bacterial Carpet
MJ Kim and KS Breuer, JFE, 2007
Active bacterial carpet in the microchannel (1 micron dia. fluorescence bead motion)
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Autonomous Bacterial Pumping System
• Coat surface of racetrack with Serratia marcesens using flow-deposited carpet
• Seed with 500 nm fluorescent particles
• Pumping velocities ~ 10 m/sec in the racetrack microchannel
PD
F-10 -5 0 5 10 15 20 250
0.02
0.04
0.06
0.08
0.1
0.12
0.14
2 mm
200 m
50 m
1.6 mm
Channel Blockedby Glue (RTV)
Streamwise Drift Velocity [ m/s]
MJ Kim and KS Breuer, APS DFD Meeting. 2004
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Fully Developed Pumping Velocity
Pumping Enhancements in the Open System:
1) Glucose (Food) Effects
2) Geometric Effects
Flagellar Motor Acitivities
Large-scale Self-coordinations
Various Effects on Bacterial Microfluidic Pumps
MJ Kim and KS Breuer, PNAS, In review. 2007
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Autonomous Bacterial Transportation System
Micro-Barges:
- Fill Factor: 90 ~ 95 %
- Typical Velocity : ~ 5 m/s
- Chemotaxis, Phototaxis, and Aerotaxis
PDMS barge
Glass substrate
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Microbarges in Motion
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Engineered Bacterial Systems
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Cell Patterning Using Colloidal Lithography
DK Yi, MJ Kim, et al. Biotech. Lett, 2006
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Conclusions
ACKNOWLEDGEMENTS:
E. Steager (Ph.D), R. Mulero (Ph.D), C.-B Kim (PostDoc), C. Naik (UG), J. Patel (UG), L. Reber (UG), S. Bith (UG).
Kenny Breuer, Tom Powers (Brown), Howard Berg, Linda Turner (Harvard), Nick Darnton (U.Mass), MunJu Kim (U.Pittsburgh).