Insulator Based Dielectrophoresis Dielectrophoresis ( (iDEP iDEP): - - PowerPoint PPT Presentation

Alfredo M. Alfredo M. Morales Moralesa

a, William D.

, William D. Munslow Munslowb

b,

, Scott M. Scott M. Maurer Maurerb

b, Blake A.

, Blake A. Simmons Simmonsa

a

a a Sandia National Laboratories, Livermore, CA 94550

Sandia National Laboratories, Livermore, CA 94550

b b Lockheed Martin MS2, Manassas, VA 20110

Lockheed Martin MS2, Manassas, VA 20110 Authorized for Unlimited Public Release by SNL SAND2009 Authorized for Unlimited Public Release by SNL SAND2009-

• 3326C

3326C And by LMC BAL200902002 And by LMC BAL200902002-

• 02

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NBC 2009 Symposium NBC 2009 Symposium Jyv Jyvä äskyl skylä ä, Finland , Finland 8 8 – – 11 June 2009 11 June 2009

Insulator Based Insulator Based Dielectrophoresis Dielectrophoresis ( (iDEP iDEP): ):

A Tool to Detect, Separate and Concentrate A Tool to Detect, Separate and Concentrate Aerosolized Biological Particles Aerosolized Biological Particles

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Biological Sensor Requirements Biological Sensor Requirements

• Requirements and Performance Measures:

Requirements and Performance Measures:

Affordability Affordability -

• Early detection

Early detection -

• Low false alarm rate

Low false alarm rate

• Current Biological Detection System Deficiencies:

Current Biological Detection System Deficiencies:

Expensive Expensive -

• Hours/days for analysis

Hours/days for analysis -

• Sensitivity

Sensitivity vs vs false alarm rate false alarm rate

• Desirable Characteristics of Typical Detect

Desirable Characteristics of Typical Detect-

• to

to-

• Warn Sensor:

Warn Sensor:

Spider Charts: metrics shown by length of legs Spider Charts: metrics shown by length of legs

Response Time (min) FAR (per year) Maintenance Interval (wks) Size (m3) Operating Cost ($/year) Unit Cost ($M) Detection Confidence

100 1 2 1000 100 1000 500 98 1 10 .5 4 .1 . 5 1 . 5 1 . 4 95 90 1 . 1 1 ,

Sensitivity (ACPLA)

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Response Time (min) FAR (per year) Maintenance Interval (wks) Size (m3) Operating Cost ($/year) Unit Cost ($M) Detection Confidence

100 1 2 1000 100 1000 500 98 1 10 .5 4 .1 . 5 1 . 5 1 . 4 95 90 1 . 1 1 ,

Sensitivity (ACPLA)

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Response Time (min) FAR (per year) Maintenance Interval (wks) Size (m3) Operating Cost ($/year) Unit Cost ($M) Detection Confidence

100 1 2 1000 100 1000 500 98 1 10 .5 4 .1 . 5 1 . 5 1 . 4 95 90 1 . 1 1 ,

Sensitivity (ACPLA)

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Ideal Biological Detection System: Ideal Biological Detection System:

• Low cost, low consumables and simple

Low cost, low consumables and simple

• peration
• peration
• Operational flexibility: Configurable as either

Operational flexibility: Configurable as either detect detect-

• to

to-

• warn or as a detect

warn or as a detect-

• to

to-

• treat system

treat system

• Expensive bioassays may be used if

Expensive bioassays may be used if inexpensive orthogonal triggers limit the inexpensive orthogonal triggers limit the number of times bioassays are run number of times bioassays are run

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Why an Why an iDEP iDEP Approach? Approach?

Flow 500 VDC 750 VDC 1000 VDC

+

• Flow

Separation by size: 2 μm (green) and 1 μm (red) Time Time Bacillus subtilis spores (red) trapped. 1 μm polystyrene beads (green) pass.

iDEP Selectively Traps Particles and May be Used as a Trigger iDEP Selectively Traps Particles and May be Used as a Trigger

Lapizco-Encinas, B. H.; Simmons, B.A., et al. , Anal.

• Chem. 2004, 76, 1571-1579

+

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iDEP iDEP Can Concentrate Bacteria Selectively Can Concentrate Bacteria Selectively

• iDEP

iDEP Separation of Live B. Separation of Live B. subtilis subtilis and B. cereus (similar bacteria) and B. cereus (similar bacteria)

• Mechanism: Different Size and Shape

Mechanism: Different Size and Shape

• Also Separated E. coli, B. cereus, B.

Also Separated E. coli, B. cereus, B. subtilis subtilis, B. , B. megaterium megaterium, B. , B. subtilis subtilis spores, TMV virus, and polystyrene particles of various sizes spores, TMV virus, and polystyrene particles of various sizes

10 20 30 40 50 60 70 80

• E. coli
• B. megaterium
• B. subtilis

B.cereus Minimum E required for DEP trapping (V/mm) medium conductivity = 2.2 microS/mm medium conductivity = 10.4 microS/mm Solution Conductivity=2.2microS/cm Solution Conductivity=2.2microS/cm Solution Conductivity=10microS/cm Solution Conductivity=10microS/cm

Selectively trapping B. cereus (green) while passing B. subtilis (red)

• E. coli < B. megaterium < B. subtilis < B. cereus

Lapizco-Encinas, B. H.; Simmons, B.A., et al. , Electrophoresis 2004, 25,1695-1704.

• +

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iDEP iDEP is a Physically Selective Phenomenon is a Physically Selective Phenomenon

• DEP: Motion of Conductive Object Toward

DEP: Motion of Conductive Object Toward (away from) High Electric Field (away from) High Electric Field

– – Along electric field gradients Along electric field gradients – – Nonlinear in the applied electric field Nonlinear in the applied electric field

• DEP Force Proportional to:

DEP Force Proportional to:

– – Particle volume Particle volume – – Conductivity + Conductivity + polarizability polarizability difference difference between particle and liquid between particle and liquid

• +

+ +

+

Net movement

Lower charge density Electrode Electrode Nonuniform electric field

• +

+ +

+

• Net movement

Higher charge density Lower charge density Electrode Electrode Nonuniform electric field

• DEP Force depend on:

DEP Force depend on:

– – Conductivity of the particle, Conductivity of the particle,  p

p

– – Conductivity of the medium, Conductivity of the medium,  m

m

– – Size of the particle, Size of the particle, r r

   

2 3

~ , ~ Re 2 E f r F

m p m

• DEP

     

   

m p m p m p

f       2 ) , ( Vdc, For   

iDEP iDEP Uses Insulating Post Uses Insulating Post Features to Shape E Features to Shape E-

• field

field

Sabounchi, P.; Morales, A.M. et al. Biomed Microdevices, 2008, 10, 661–670.

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iDEP iDEP with Pressure Driven Flow with Pressure Driven Flow

• Large Sample Volumes Must be Processed for Many Clinical Applica

Large Sample Volumes Must be Processed for Many Clinical Applications tions

• Only Pressure Driven Flow can Process Such Volumes

Only Pressure Driven Flow can Process Such Volumes

Electrokinetic Flow Profile Pressure Driven Flow Profile iDEP Force Force due to flow

• Potential Attributes of Biological

Potential Attributes of Biological Sensor Incorporating Sensor Incorporating iDEP iDEP: :

– – Affordability: Affordability: iDEP iDEP trigger for bioassay trigger for bioassay – – Early Detection: Response within Early Detection: Response within seconds of turning on seconds of turning on – – Low False Alarm Rate: Orthogonal Low False Alarm Rate: Orthogonal iDEP iDEP trigger trigger



   F U

particle particle

) (u

• dt

du m 

) ( E E E U u     

DEP EK particle

 

Fluid Flow Fluid Flow Electrokinetic Electrokinetic Velocity Velocity Dielectrophoretic Dielectrophoretic Velocity Velocity

A Significant Amount of Work A Significant Amount of Work Done in the Lab. Done in the Lab. What Will it Take to Field What Will it Take to Field iDEP iDEP? ?

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Micro Micro-

• molded

molded iDEP iDEP Zeonor Zeonor Chips Chips

• Polymer

Polymer iDEP iDEP Chips: Chips:

– – Low Cost & Ease of Manufacture Low Cost & Ease of Manufacture – – Amenable to Large Scale Production Amenable to Large Scale Production

R1 R2 R3 R4 x y

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Chip Surface and Buffer Chemistry Is Critical Chip Surface and Buffer Chemistry Is Critical

• Surface & buffer chemistry enable repeatable trapping with press

Surface & buffer chemistry enable repeatable trapping with pressure driven flow ure driven flow

• MOPA Treatment Lowers Contact angle,

MOPA Treatment Lowers Contact angle, 90 90   2 2° ° w/o MOPA w/o MOPA vs vs 65 65   4 4° ° w/ MOPA w/ MOPA

• Background Buffer pH Adjusted to 8

Background Buffer pH Adjusted to 8 While Keeping Conductivity Low While Keeping Conductivity Low

• Compatible with TWEEN used in

Compatible with TWEEN used in Aerosol Sample Collector Aerosol Sample Collector

O O

8

O OH O O OR OH

+ + NaIO4

MOPA (Methoxy polyethylene glycol acrylates) “Chip polymer” Functionalized “chip polymer”

With MOPA Without MOPA

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Waste Manifold Waste Manifold

System Layout and Architecture System Layout and Architecture

Aerosol Collector Aerosol Collector Test Sample Test Sample iDEP iDEP Chip Chip Background Buffer Background Buffer

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Robust Automated Robust Automated iDEP iDEP System System

• Designed a Robust System Capable of Operating in the Field

Designed a Robust System Capable of Operating in the Field

• Both Software & Hardware were Simplified and Optimized with

Both Software & Hardware were Simplified and Optimized with Reliability and Safety in Mind Reliability and Safety in Mind

• Prototype Field Test Unit Fabricated, Integrated and Tested

Prototype Field Test Unit Fabricated, Integrated and Tested

Modular iDEP chip holder

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Commercial microfluidic connectors

Robust Automated Robust Automated iDEP iDEP System System

• Designed a Robust System Capable of Operating in the Field

Designed a Robust System Capable of Operating in the Field

• Both Software & Hardware were Simplified and Optimized with

Both Software & Hardware were Simplified and Optimized with Reliability and Safety in Mind Reliability and Safety in Mind

• Prototype Field Test Unit Fabricated, Integrated and Tested

Prototype Field Test Unit Fabricated, Integrated and Tested

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Electronics, HVS, and aerosol collector integrated

Robust Automated Robust Automated iDEP iDEP System System

• Designed a Robust System Capable of Operating in the Field

Designed a Robust System Capable of Operating in the Field

• Both Software & Hardware were Simplified and Optimized with

Both Software & Hardware were Simplified and Optimized with Reliability and Safety in Mind Reliability and Safety in Mind

• Prototype Field Test Unit Fabricated, Integrated and Tested

Prototype Field Test Unit Fabricated, Integrated and Tested

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Rack mountable packaging

Robust Automated Robust Automated iDEP iDEP System System

• Designed a Robust System Capable of Operating in the Field

Designed a Robust System Capable of Operating in the Field

• Both Software & Hardware were Simplified and Optimized with

Both Software & Hardware were Simplified and Optimized with Reliability and Safety in Mind Reliability and Safety in Mind

• Prototype Field Test Unit Fabricated, Integrated and Tested

Prototype Field Test Unit Fabricated, Integrated and Tested

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Dedicated control and data logging software

Robust Automated Robust Automated iDEP iDEP System System

• Designed a Robust System Capable of Operating in the Field

Designed a Robust System Capable of Operating in the Field

• Both Software & Hardware were Simplified and Optimized with

Both Software & Hardware were Simplified and Optimized with Reliability and Safety in Mind Reliability and Safety in Mind

• Prototype Field Test Unit Fabricated, Integrated and Tested

Prototype Field Test Unit Fabricated, Integrated and Tested

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Sample Automated Sample Automated iDEP iDEP Run Run

Free Flow (0 V) Free Flow (0 V) Trapping (1600 V) Trapping (1600 V) Release (0 V) Release (0 V)

Time

Fluorescently labeled 2 μm polymer beads (106 particles/ml)

• +

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Fluorescence Detector is Immune to Fluorescence Detector is Immune to iDEP iDEP Artifacts Artifacts

• Off

Off-

• chip, Capillary

chip, Capillary-

• based

based Conductivity and Absorbance Conductivity and Absorbance Detectors are Affected by Detectors are Affected by iDEP iDEP-

• trapping Related Artifacts

trapping Related Artifacts

• Fluorescence Detection not

Fluorescence Detection not Affected by Artifacts. On Affected by Artifacts. On-

• chip

chip Fluorescence Detection Senses: Fluorescence Detection Senses:

• Trapping & concentration between

Trapping & concentration between posts posts

• Trapped particles shortly after

Trapped particles shortly after trapping voltage is turned on trapping voltage is turned on

• System response in seconds

System response in seconds

• Biological particles by detecting

Biological particles by detecting native fluorescence native fluorescence

• Chip Holder Design Becomes

Chip Holder Design Becomes Complex Complex

Fluorescently Labeled Polymer Beads (106 particles/ml)

Fluorescence Analysis Areas Fluorescence Imaging

20 40 60 80 100 Time(sec) Trapping Voltage (v Fluorescence (au Trapping Voltage Fluorescence

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Off Off-

• Chip Capillary Based Fluorescence Detector

Chip Capillary Based Fluorescence Detector

• These LIF Detectors Have Been Used for Many

These LIF Detectors Have Been Used for Many Years Both in the Lab and in the Field at Years Both in the Lab and in the Field at Various Locations Various Locations

• The Detector Units were Developed to

The Detector Units were Developed to be be Miniature, Rugged, & Miniature, Rugged, & Consume Little Power Consume Little Power

• They Measure Approximately 2.5 X 3 X 2 in,

They Measure Approximately 2.5 X 3 X 2 in, require require 5.3 volts, < 100 milliamps, Excluding 5.3 volts, < 100 milliamps, Excluding the Laser Source the Laser Source

• Detector has been used with 405, 488, 532, 635,

Detector has been used with 405, 488, 532, 635, and 650 nm Excitation Sources and 650 nm Excitation Sources

Off-Chip Miniaturized Laser Induced Fluorescence Detector Unit

Renzi, R. et al, Anal. Chem., 2005, 77(2), pp 435–441

Embedded Capillary Fluorescence Provides Real-Time Detection Embedded Capillary Fluorescence Provides Real-Time Detection

18 50 100 150 200 250 300 350 400 Time (sec) Trapping Voltage (v) Average Counts Per Second Trapping Voltage Avg CPS 50 100 150 200 250 300 350 400 Time (sec) Trapping Voltage (v) Average Counts Per Second

Trapping Voltage Avg CPS

Fluorescence Indicates Trapping and Release Fluorescence Indicates Trapping and Release

• No Redesign of Chip Holder Needed

No Redesign of Chip Holder Needed

• Fluorescence CPS Changes as Soon as Trapping Starts

Fluorescence CPS Changes as Soon as Trapping Starts

• Trapping is Seen a Minute after Voltage is Applied

Trapping is Seen a Minute after Voltage is Applied vs vs Concentration! Concentration!

• A Slight Delay is Introduced by Locating Sensor Off

A Slight Delay is Introduced by Locating Sensor Off-

• chip

chip

Fluorescently Labeled Polymer Beads (106 particles/ml) Fluorescently Labeled Bacillus atrophaeus (106 particles/ml) Trapping Trapping

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Summary Summary

• iDEP

iDEP Has Several Features and Can: Has Several Features and Can:

– – Selectively trap and concentrate particles Selectively trap and concentrate particles – – Be used as a trigger in a cost effective, fast Be used as a trigger in a cost effective, fast biodetector biodetector

• Automated, Prototype

Automated, Prototype iDEP iDEP Field Unit has been Built, Field Unit has been Built, Integrated and Tested Integrated and Tested

• Embedded Fluorescence Detection has been shown to

Embedded Fluorescence Detection has been shown to Sense Trapping and Concentration in < 1 minute After Sense Trapping and Concentration in < 1 minute After Applying Applying iDEP iDEP Voltage Voltage

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Questions? Questions?

Acknowledgments Acknowledgments

Sandia National Labs: Sandia National Labs:

• Tim

Tim Shepodd Shepodd

• Josh Whaley

Josh Whaley

• Mark D. Zimmerman

Mark D. Zimmerman

• Benjamin Van

Benjamin Van Blarigan Blarigan

• Karen L.

Karen L. Krafcik Krafcik

• April

April Nissen Nissen

• Joanne

Joanne Volponi Volponi

• Ron

Ron Renzi Renzi

• Nicholas

Nicholas Mascarenhas Mascarenhas

Previous SNL and LMC funding Previous SNL and LMC funding

Lockheed Martin: Lockheed Martin:

• Tyesia

Tyesia Pompey Pompey

• Stephanie Groves

Stephanie Groves

• Frank

Frank Rotondo Rotondo

• Robert

Robert D D’ ’Italia Italia