MEMS for Bio Applications MEMS for Bio Applications October 14, - - PDF document

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MEMS for Bio Applications MEMS for Bio Applications October 14, 2003 Kim, Yong-Kweon School of Electrical Engineering and Computer Science Seoul National University Lab. for Micro Sensors and Actuators, SNU since 1992 Contents Contents

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SLIDE 1
  • Lab. for Micro Sensors and Actuators, SNU since 1992

MEMS for Bio Applications MEMS for Bio Applications

October 14, 2003

Kim, Yong-Kweon

School of Electrical Engineering and Computer Science Seoul National University

Kim, Yong-Kweon 2/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Contents Contents

  • MEMS Applications
  • Bio Applications
  • Micro Array: Peptide Micro Array
  • Bead Affinity Chromatography Chips
  • Enzyme Reaction in a Micro Fluidic System
  • Bio Measurement : Lens and Scanner
  • Summary
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SLIDE 2
  • Lab. for Micro Sensors and Actuators, SNU since 1992

Applications of MEMS Applications of MEMS

Kim, Yong-Kweon 4/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Applications of MEMS Applications of MEMS Applications of MEMS

  • Pressure sensors
  • Accelerometer
  • Gyroscope
  • Digital Micromirror Device
  • I nkjet Head
  • Optical Switch
  • Pressure sensors
  • Accelerometer
  • Gyroscope
  • Digital Micromirror Device
  • I nkjet Head
  • Optical Switch
  • MEMS has been developed as a break-through technology.
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SLIDE 3

Kim, Yong-Kweon 5/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Conventional vs. MEMS I nertial Conventional vs. MEMS I nertial Measurement Units Measurement Units

Mass : 1587.5 grams Size : 15 cm x 8 cm x 5 cm Pow er : 35 W Survivability : 35 G’s Cost : $ 20,000 Mass : 10 grams Size : 2 cm x 2 cm x 0.5 cm Pow er : ~ 1 mW Survivability : 100 kG’s Cost : $ 500 I nertial Measurement Unit

Conventional MEMS

From DARPA

  • The merits of MEMS are to be small, light, cheap, multi-functional and

integrated with mechanical and electronic components.

Kim, Yong-Kweon 6/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Mass Spectrograph on a Chip Mass Spectrograph on a Chip

  • Mass spectrograph on a chip will be integrated with vacuum pumps,

ionizer, an ion detector array, and control electronic circuits.

  • The MS on a chip enables potable measurement units and point-of-care

will be available.

I on Detection Array Processor Support Electronics I onizer Dual Filter Magnet Filter I on Optics Vacuum Pumps Slits Memory Sample Gas I nput

$ 17,000 7 0 kg 1,200 W 20,000 cm3 Mass Spec Chip $ 20 0.2 kg 0.5 W 3 cm3 Conventional MEMS From DARPA

  • BioMEMS provides new methodologies to the biotechnology.
  • For examples, lab-on-a-chips, Affymetrix DNA chips, and high throughput

screening chips continue to evolve the biotechnology.

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SLIDE 4
  • Lab. for Micro Sensors and Actuators, SNU since 1992

Bio Applications : Peptide Micro Array Bio Applications : Peptide Micro Array Synthesized using Synthesized using Micromirror Micromirror Array Array

Kim, Yong-Kweon 8/ 25

  • Lab. for Micro Sensors and Actuators, SNU
  • DNA micro arrays
  • DNA sequencing assay
  • Microfabrication

(a) Contact printing (b) Inkjet printing (c) In situ fabrication using inkjet (d) In situ fabrication using photolithography

  • Merits
  • One chip assay
  • Performance is improved
  • Assay time and cost are

reduced

  • Easy manipulation

DNA Micro Array DNA Micro Array

DNA -double helix strands

DNA broken into fragments and tagged Base pair being investigated

Segment from cell being tested Probes that test for: Weak bonds Strong bond

Enlarged view of four fields Field Gene chip

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SLIDE 5

Kim, Yong-Kweon 9/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Making Micro Arrays Making Micro Arrays

  • I n situ fabrication using

photolithography

  • Light at 365nm is shone through a mask.
  • The light releases the capping chemical,

exposing parts of the substrate.

  • A solution is then washed over the substrate.
  • The nucleotides attach to the unprotected

sites, adding their own capping layer.

  • The process is repeated, building up

sequences of DNA.

  • Making micro arrays
  • Just printing

(a) Contact printing (b) Inkjet printing

  • In situ fabrication

(c) In situ fabrication using inkjet (d) In situ fabrication using photolithography

I n situ fabrication using inkjet

Repeat

I n situ fabrication using Photolithography I nkjet printing Contact printing

Wetted pin Light at 365 nm Mask Photosensitive chemical Nucleotide with photosensitive chemical Nucleotide Nozzle Nozzle Substrate

  • Suitable to mass production with

the same sequence.

Kim, Yong-Kweon 10/ 25

  • Lab. for Micro Sensors and Actuators, SNU

UV illuminator Micromirror Off-state Micromirror On-state Micro fluidic systems Port connected to a biochip reagent Wasting port Observation camera Reagent delivery port Reaction chamber

Schematic drawing of biochip fabrication system

Synthesis of Peptide Micro Array using Synthesis of Peptide Micro Array using Micromirror Micromirror Array Array

Mircromirror : 50 ㎛ x50 ㎛ Mirror array-16x16 3 x 3 units operation (1 unit : 2 x 2 mirrors) Photoresist(AZ1512) patterns produced by micromirror array

SoEE

X distance a b c de f g h Proje ction time[ min] 2 4 6 8 1 1 2 1 4 1 6 Dut y cycl e of mir ror = 0.6 8

Peptide micro array

Y distance Line 3 (I HPQG) Line 1 (Biotin) Line 2 (HPQI G) Line 4 (I GHPQ)

Y.-K. Kim and Y.-S. Lee, SNU

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SLIDE 6

Kim, Yong-Kweon 11/ 25

  • Lab. for Micro Sensors and Actuators, SNU

MMA Flow system for peptide/ligand synthesis Reaction chamber Outlet Slide chip UV light illumination UV Imaging optics (lens, filter, stop) Inlet

MMA Systems for Peptide Synthesis on a Chip MMA Systems for Peptide Synthesis on a Chip

  • Virtual mask pattern

generator

  • Micromirror array(MMA)
  • Virtual mask pattern

generator

  • Micromirror array(MMA)
  • UV imaging optics for MMA

photolithography

  • UV imaging optics for MMA

photolithography

  • Flow cell for surface

synthesis chemistry

  • Reagent delivery to the

flow cell

  • Flow cell for surface

synthesis chemistry

  • Reagent delivery to the

flow cell

Y.-K. Kim, SNU

Kim, Yong-Kweon 12/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Micromirror array

UV illuminator Micromirror On-state Micromirror Off-state Selective photolithography A11 A12 A13 A21 A2

2 A2 3

A31 A3

2 A3 3

A A’

Fabricated biochip

Amino acids G: Glycine I : I soleucine Q : Glutamine P : Proline H : Histidine Amino acids G: Glycine I : I soleucine Q : Glutamine P : Proline H : Histidine

Peptide Array Synthesis on a Chip using MMA and Peptide Array Synthesis on a Chip using MMA and Protein Binding Assay Protein Binding Assay

Glass spacer N H NVOC spacer N H NVOC i) UV(365nm, 30mW/ cm

2)

UV ii) 5mM NVOC- G BOP,HOBt in DMF i) UV(365nm, 30mW/ cm

2)

ii) 5mM NVOC- I BOP,HOBt in DMF NVOC NVOC UV I NVOC NVOC ii) FI TC

  • streptavidin

in 50mM phosphate buffer ( pH= 7.4 ) i) 0.5% (w/ v) BSA in 50mM phosphate buffer ( pH= 7.4 ) spacer N H NVOC spacer N H NVOC spacer N H NVOC spacer N H NVOC spacer N H NVOC spacer N H NVOC Glass spacer N H NVOC spacer N H G spacer N H NVOC spacer N H G Glass spacer N H NVOC spacer N H NVOC spacer N H G spacer N H G spacer N H NVOC spacer N H NVOC spacer N H G spacer N H G Glass spacer N H NVOC spacer N H G spacer N H NVOC spacer N H G Glass spacer N H NVOC spacer N H NVOC spacer N H G spacer N H G spacer N H NVOC spacer N H NVOC spacer N H G spacer N H G I Q Q P P H G I H I P H Glass spacer N H Biotin spacer N H G spacer N H Q spacer N H G I Q Q P P H G I H I P H Glass spacer N H Biotin spacer N H G spacer N H Q spacer N H G Glass spacer N H Biotin spacer N H Biotin spacer N H G spacer N H G spacer N H Q spacer N H Q spacer N H G spacer N H G Total ; 9 photolithography processes

Y.-S. Lee, SNU

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SLIDE 7

Kim, Yong-Kweon 13/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Ligand Ligand I nteraction using MMA I nteraction using MMA

Assay on a chip Assay on a chip

Fluorescence assay v s Enzyme assay Sensitivity & S/N One chip analysis v s 4 times ELISA with sophisticated controlled condition

Biotin HPQIG IHPQG IGHPQ 0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 Opitical Density[a.u.]

Assay on beads Assay on beads

Error range Assay time( labor) reduced ( < 1/ 7days) , Convenience( automated synthesis by MMA systems), I mprovement of assay reliability by comparison on a chip Assay time( labor) reduced ( < 1/ 7days) , Convenience( automated synthesis by MMA systems), I mprovement of assay reliability by comparison on a chip

Bg Line 1 Line 2 Line 3 Line 4

20 30 40 50 60 70 80 90 Intensity of fluorescence[au]

Error range Biotin HPQIG I HPQG IGHPQ

Line 3 Line 1 Line 2 Background Line 4 Line 3 Line 1 Line 2 Background Line 4 X distance Y distance

Y.-S. Lee, SNU

Kim, Yong-Kweon 14/ 25

  • Lab. for Micro Sensors and Actuators, SNU

affinity assay of streptavidin < 1 day ~ 7 days Assay time (labor) 3 sequences x penta peptide = 15 times 15 μmol 15 times x 150 μmol = 2250 μmol Reagent quantity (Amino acids) 100 mg resins for each penta peptide for 3 kinds

  • f sequence

G:Glycine, I : I soleucine, Q: Glutamine, P: Proline, H: Histadine Application example 1μmol / reaction 5 mM solution in 200 ㎕ flow cell 1 nmol/ cm 2 On surface(chip) 150 μmol / reaction Required amino acid resin 500 mg Quantity 0.1 mmol/ g Surface density Comments On resin(bead) NH2 GI QPH GQPHI QPHGI GI QPH GQPHI QPHGI NH2

Merits of the Synthesis using MMA Merits of the Synthesis using MMA

Y.-K. Kim and Y.-S. Lee, SNU

  • The MMA system provides the flexibility on biopattern design and fast turn-

around analysis.

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SLIDE 8
  • Lab. for Micro Sensors and Actuators, SNU since 1992

Bio Applications : Bead Affinity Bio Applications : Bead Affinity Chromatography Chips Chromatography Chips

Kim, Yong-Kweon 16/ 25

  • Lab. for Micro Sensors and Actuators, SNU

(a) I njection biosamples (b) Concentration & separation of target protein (c) Elution of target protein using UV cleavage (d) Analysis of target protein using mass spectrometry or fluorescence

Bead Affinity Chromatography (BAC) Bead Affinity Chromatography (BAC)

Y.-K. Kim and Y.-S. Lee, SNU

  • BAC chips are based on the idea using a large surface area of be ads to

analyze bio target molecules in a low concentration.

  • Silicon and glass based micromachining makes the reaction chamber

packed micro beads and micro channels.

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SLIDE 9

Kim, Yong-Kweon 17/ 25

  • Lab. for Micro Sensors and Actuators, SNU

29.3 mm 36.83 m m

Reactor depth 50 ㎛ Total volume : 1 ㎕ Micro Beads Filter 500㎛ Left : magnetic bead Right : CutiCore bead Reactor depth 100 ㎛ Total volume : 0.4 ㎕

Micro Micro Bead Affinity Chromatography Chips Bead Affinity Chromatography Chips

Y.-K. Kim , B.-G. Kim and Y.-S. Lee, SNU

Kim, Yong-Kweon 18/ 25

  • Lab. for Micro Sensors and Actuators, SNU
  • Packed bead (40 ㎕ of 0.33 % bead suspension), BSA blocking (20 min),
  • Sample injection and reaction (30 min), room temperature
  • Streptavidin-FI TC (1 - 50 ㎍/㎖) / PB (pH 7.4)
  • UV irradiation for 30 min and elution with PB
  • Packed bead (40 ㎕ of 0.33 % bead suspension), BSA blocking (20 min),
  • Sample injection and reaction (30 min), room temperature
  • Streptavidin-FI TC (1 - 50 ㎍/㎖) / PB (pH 7.4)
  • UV irradiation for 30 min and elution with PB

2 4 6 8 1 1 2 1 4 1 6 1 8 1 2 3 4 5 6

Streptavidin-FI TC (㎍/ ㎖) Peak area (a.u.)

y = 270.3x + 1962.5 R2 = 0.9818

3 60 90 120 150 180 500 1000 1500 2000 2500

Fluorescence(a.u.) time(s)

3 60 90 120 150 180 500 1000 1500 2000 2500 31.6

Fluorescence(a.u.) time(s)

3 60 90 120 150 180 500 1000 1500 2000 2500

Fluorescence(a.u.) time(s)

30 60 90 120 150 180 500 1000 1500 2000 2500

Fluorescence(a.u.) time(s)

30 60 90 120 150 180 500 1000 1500 2000 2500

1㎍/ ㎖ 10㎍/ ㎖ 5㎍/ ㎖ 25㎍/ ㎖ 50㎍/ ㎖ time(s) Fluorescence(a.u.)

Chromatogram of eluted protein

Quantitative Analysis Quantitative Analysis Streptavidin Streptavidin-Biotin Model System Biotin Model System

Y.-K. Kim and Y.-S. Lee, SNU

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SLIDE 10

Kim, Yong-Kweon 19/ 25

  • Lab. for Micro Sensors and Actuators, SNU

1.1311.961( LPINALSNSLLR) 2.1511.846(YLFNWAVKTKLK) 3.1835.717( WEYVLLLFLLLADAR) 4.2234.996(LNAACNWTRGERCDLEDR) 5.2585.975(SLTERLYIGGPLTNSKGQNCGYR) 6.2651.06(CRASGVLTTSCGNTLTCYLKASAACR) 7.2779.324(ALDCQIYGACYSIEPLDLPQIIER)

100 - 200 fmol/ ㎕ of HCV RNA polymerase was detected

1000 2000 3000 1 2 3 4 5 6 7

A.Normal serum (21.woman)

Relative Intensity

B.Hepatitis C patient

  • Mass analysis
  • Fluorescence detection

HCV RNA Polymerase-FI TC Mixture HSA-FI TC Ovalbumin-FI TC Streptavidin-FI TC

50 100 150 2 2 5 200 400 600 800

Fluorescence intensity (arb. unit) Time (s)

HCV RNA Polymerase Detection and Analysis HCV RNA Polymerase Detection and Analysis by MALDI by MALDI -TOF & Fluorescence TOF & Fluorescence

Y.-K. Kim , B.-G. Kim and Y.-S. Lee, SNU

  • To show the feasibility, we tested the HCV RNA polymerase detection.
  • With MALDI and fluorescence detection, HCV RNA polymerase are

detected, even in cocktailed samples.

  • Lab. for Micro Sensors and Actuators, SNU since 1992

Bio Applications : Enzyme Reaction in a Bio Applications : Enzyme Reaction in a Micro Fluidic System Micro Fluidic System

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SLIDE 11

Kim, Yong-Kweon 21/ 25

  • Lab. for Micro Sensors and Actuators, SNU

      +       + = hw w hw w h P

FC PDMS

θ γ θ γ cos 2 cos

Hydrophobic PDMS Hydrophobic FC film A negative pressure indicates a repellent force at the solid-liquid-gas interface.

Variable Metering using a T Variable Metering using a T-shaped shaped Microinjecor Microinjecor with Sidewall with Sidewall Microchannel Microchannel Array Array

Sidewall microchannel array (Hydrophobic valve) I njection channel Hydrophilic glass Hydrophobic PDMS Hydrophobic FC pattern Metering channel Scale bar

  • Nanoliter droplets are controlled using the surface property such as

hydrophobic and hydrophilic.

Kim, Yong-Kweon 22/ 25

  • Lab. for Micro Sensors and Actuators, SNU

I njection channel 2 Hydrophilic glass Pneumatic control channel 1 Air venting channel 3 Air venting channel 4 PDMS channel network Metering region Hydrophobic FC patterns Reaction region Scale bar

  • Nanolitermeasuring step

Sidewall microchannel array (Hydrophobic valve “ON”) Effective length (30 nl)

  • Flow direction control step

Fabricated device 30 s ( b) Enzyme side FDG side ( c) A

A´

7 min 30 s

( a)

FDG Enzyme I nitial state Reaction on a micro fluidic system Flow direction Hydrophilic surface Hydrophobic surface

Enzyme Reaction in a Micro Fluidic System Enzyme Reaction in a Micro Fluidic System

fluorescein di-ß- galactopyranoside

Y.-K. Kim and B.-G. Kim

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SLIDE 12
  • Lab. for Micro Sensors and Actuators, SNU since 1992

Bio Applications : Measurements using Bio Applications : Measurements using Optical MEMS Components Optical MEMS Components

Kim, Yong-Kweon 24/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Micro prism (PMMA) fabricated using hot embossing Fiber fixer & micro lens Scanning mirror Micro prism Collimating lens Location of protein chip Photodetector

Schematic drawing of Bio Measurements

2D micro scanner Mirror : 3.5 mm x 3.5 mm Micro scanning mirror Micro spring Lissajous pattern illuminated using micro scanner

  • 1. 2D surface scan (silicon spring)
  • 2. 1:2 freq. ratio (silicon spring)
  • 3. 1:2 freq. ratio (polymer spring)
  • 4. 2:5 freq. ratio (polymer spring)

Resonance of movable frame at 150Hz

Micro lens (PMMA)

PMMA Glass Resonance of mirror at 380Hz Laser spot (off state) Scanning line ( driving movable frame only) Scanning line (driving mirror

  • nly)

Bio Measurements using Optical MEMS Bio Measurements using Optical MEMS

Y.-K. Kim and H. –S. Kim

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SLIDE 13

Kim, Yong-Kweon 25/ 25

  • Lab. for Micro Sensors and Actuators, SNU

Summaries Summaries

  • MEMS technology provides the strong methodologies to

biotechnology, life science and chemical engineering.

  • Nanotechnology will also be a break-through technology

to biotechnology.

  • Acknowledgements
  • These researches have been supported by MOST (Ministry of

Science and Technology) and KOSEF (Korea Science and Engineering Foundation).

  • Co workers
  • Prof. Yoon-Sik Lee, Prof. Byung-Gee Kim (SNU), Prof. Hoseong

Kim (ChungAng Univ.)

  • Post Doc. Researchers : Dr. Kooknyung Lee, Dr. Sangho Lee, Dr.

Chang-Soo Lee (SNU).

  • Graduate Students: Minsoo Kim, Suhyung Cho, Woojae chung

(SNU), Yongyoon Kim (ChungAng Univ.).