Laboratory-on-Chip : Purpose · Laboratory-on-Chip : Purpose • A multidisciplinary approach to miniaturize biological assays or procedures in ... EZ Smart Monitoring Laboratory-on-Chip

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March 2011

GBM8320 - Dispositifs Médicaux Intelligents 2

Laboratory-on-Chip : Purpose • A multidisciplinary approach

to miniaturize biological assays or procedures in analytical chemistry.

• To embed the required equipments for performing the biological assays in a single chip for single purpose.


• Low sample consumption;

• Fast biochemical reactions & Miniaturization;

• Point-of-care.

Laboratory-on-Chip : Advantage

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Laboratory-on-Chip : CMOS technology

•  Low voltage, and low power;

•  Addressable sensing or manipulating sites;

•  Standard technology.


I.  Introduction

II.  Biochemistry

III.  Microfluidic Packaging

IV.  Capacitive Sensors

V.  On-Chip Cells Detection and Manipulation.

Laboratory-on-Chip : Outline


Laboratory-on-Chip : Miniaturization

•  Low voltage E=V/d (d: 1cm to 1um, V:10kV to 1V) •  Low-sample low-cost experiment with expensive

biomaterials 1ml to 1nl •  Fast biological assays 1 hr to 1ms •  Miniaturized devices •  Home healthcare, Point-of-care monitoring, Home

diagnostic,… -------- •  Portable devices •  Miniaturization is only solution for single cell

detection : Sensitivity = DVolume/Volume

d!

+! -!

V!

E!

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Application Method Microfluidic CMOS Year Authors Cells

monitoring Capacitive

Sensor Adhesive technique 0.5um 2007 Prakash et al

Organic solvent Sensor

Core CBCM capacitive Sensor

DWFP 0.18um 2007

Ghafar-Zadeh et al

Bioluminane detection

Optical Method

Rapid Prototyping 0.18um 2007 Eltoukhy et al

DNA detection Capacitive

Sensor Rapid

Prototyping 0.5um 2006 Stagni et al

Magnetic manipulation

Array of !coils

MEMS based Procedures 0.18um 2006 Lee et al

Virus detection

Capacitive Sensor

Rapid Prototyping 0.35um 2005

Balasubramanian et al

Cell localization

Capacitive Sensor

Adhesive technique 0.35um 2004 Romani et al

Dielectrophoresis Array of Rapid 0.35um 2003 Medoro et al manipulation !electrodes prototyping

Laboratory-on-Chip : CMOS-Based !systems


Affymetrix Inc.!

Requires scanner for reading out!

EZ Smart Monitoring

Laboratory-on-Chip : Technologies

Glucose Detection DNA !array Rapid Bacterial Diagnosis

STMicro-electronics


Laboratory-on-Chip : Future Technologies

Display

Disposable needle

Sensor

Syringe Microchannel

Past Room

Laboratory

Today Desktop

Laboratory

Future Pencil-Size Laboratory

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Lab-on-Chip Information

technology

Selected bioparticle

Electric Signal

Diagnostic information

BIO

Material flow

Information flow

Micro / Nano

BioInfo

Patient Physician

Blood sample

preparation

Laboratory-on-Chip : Multidisciplinarity !

Lab-on-

Chip


• Microfluidic structure ; • Sensing layer ; •  Interface circuit : Sensor

or Manipulator.

Interface Circuit

Vdd !C

Electrode

Cell

Microchannel

Sensing Layer

Laboratory-on-Chip : CMOS-based devices!

•  On-chip non-uniform electric field;

•  Cell manipulation.


,...,C m! ! Q!

•  Low complexity capacitive sensor •  High precision optical sensor

Antigen labeled by fluorescence materials

Beads

Note: Impedometric and cantilever require removing the passivation layers

Antibody

Antigen

Laboratory-on-Chip : Optical/Capacitor sensing!

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!  Optical biosensors are becoming popular due to inherent advantages. !  The basis is enzymatic reactions, which alter the optical properties of

substances allowing to emit light upon illumination ; !  Means of detection include fluorescence, phosphorescence, chemi /

bioluminescence. -------------

!  Photometric fiber sensors - changes in reflectivity & spectrum ; !  Temperature based fiber microprobes - The luminescence of a crystal at

the probes tip is temperature dependent (-50 to 200oC) ; !  Fiber based Pressure sensors - The fiber is in a catheter and operates

by the displacement of a membrane ; !  Chemical Sensors - Use of permeable membrane on the fiber probe,

which contains a reversible indicator that responds to a chemical stimulus by absorption or luminescence.

Laboratory-on-Chip : Optical/Capacitor sensing!


SHS

SHR

SIG

RST

COL BUS

RS

RST

VDD

CS

HOR BUS

VLN

VLP

VLP

PIXEL

GLOBAL

Integration starts

Signal sampled

Pixel reset level sampled Photodiode Voltage

0V

1V

2V

3V

33 ms 66 ms

Net signal

0V

1V

2V

3V

33 ms 66 ms

Laboratory-on-Chip : CMOS Imaging!


Our reported new applications : 1) Organic Solvent Detection, 2) Bacteria Growth Monitoring, 3) Polyelectrolyte layer detection.

Cell monitoring

(charges generate dipoles).

Organic solvent detection (Chemical gas change dielectric capacitance).

DNA detection

(hybridisation generates extra

charges)

Antibody-antigen recognition

Laboratory-on-Chip : Capacitor sensing applications!

Hagleither et al, IEEE SSCJ, Dec 2002. Ghafar-Zadeh et al, CJECE, 2008.

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I.  Introduction

II.  Biochemistry






Cell Separation DNA Extraction DNA Hybridization

For performing the biological assays, several equipments are required.

Manipulation protocols (Temperature, volume, others..).

DNA (Deoxyribonucleic) is a nucleic acid that contains the genetic instructions used in the development and functioning of all known

living organisms.

Laboratory-on-Chip : Fundamental of Biochemistry!


Blood Chromosomes!

DNA Strands!

Fundamental of biological particles Blood ; Cells ; Chromosomes ; Molecules ; Genes ; DNA ; Fractioning…

•  Genetic info is stored in the cell chromosomes •  A chromosome consists of long supercoiled strands of molecules. •  Each strand is a string of DNA fragments grouped as genes, each

expressing an identifiable function of the organism.


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Sampling Concentration Purification

Amplification Detection Signal Analysis

A typical biological test through desktop, or room size systems

PCB

Microfluidic

Sensor

Analog Digital and Software Microfluidic and Temperature control system

Microfluidic needle Dielectrophoresis (no charge) Electrophoresis (charge)

P.Grodzinski & al. Motorola labs. Anal. chem. 76, P.1824, 2004

Uniform electric field

•  Dielectrophoresis (DEP) for Separation, levitation, rotation, etc.

•  Electrophoresis for molecule level preparation.


Polymerase Chain Reaction (PCR)

Centrifuge system Fragmentation

PCR

Cells

DNA

Molecules



•  Gold electrode on CMOS •  Solution is spread on electrodes for 18 h. •  Single stranded DNA molecules immobi-

lized on the electrodes. •  Unknown molecule detected through a

CMOS capacitive sensor.

Affymetrix Inc. -- On-chip DNA Detection --

LoC innovation : Gene chip. It requires: •  Optical Scanner for readout / data mining ! On-Chip Reader is needed, capacitive

sensor is a good candidate.


Fundamental of Biochemistry : DNA!•  There are four types of nucleotides corresponding to four different

bases: adenine, guanine, cytosine, and thymine (A, G, C and T). •  DNA Individual bases are hydrophobic, but strands of DNA are

quite soluble in water due to the polar backbone. •  Single-stranded DNA tends to attach (or hybridize) through weak

hydrogen bonds to another strand of complementary base pairs (G–C and A–T), forming a double strand.

•  In humans, for example, each of the 46 chromosomes is 5x400 exp (6) units long, while the single chromosome in the E. coli bacteria is 4x10 exp (6) units long.

•  Each nucleotide consists of a base (B), a sugar linkage (S) and a phosphate bridge (P).

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Fundamental of Biochemistry : Microarrays!

Ink jet

Pin Quill Pen

Pin Ring

•  Robotic DNA Arrays : dispensing artificial DNA molecules (or probes) for diagnosis purposes.

•  Photolithography method to create artificial DNA probes for diagnosis purposes (very large scale immobilized polymer array synthesis).

Light Mask

Gene-Chip

•  Modify the surface by various chemical reactions to create the required T, C, G, A..


•  An antigen is defined as a substance that can be bound by an antibody molecule through its antigen-binding sites, also called epitopes.

Signal

No Signal Anti-body

Specific Recognition

Non-specific Recognition


Antibody

Antigen


I.  Introduction

II.  Biochemistry





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Laboratory-on-Chip : Microfluidic Packaging!

Ink jet printer

http://en.wikipedia.org/wiki/Inkjet_printer http://www.imaging.org/ist/resources/tutorials/inkjet_printer.cfm


•  3D microstructures for fluidics (Silicon)

•  Various applications: microvalves, microjets, lab-on-chip, Gas Chromatography, microfluidics.

MINOS, Ljubljana, D. Resnik et al

•  Micronozzle on silicon for sampling purposes.

Laboratory-on-Chip : Microfluidic Packaging!


Microfluidic Packaging : MEMS-based techniques!

Mold casting

Mold release

Casting against mold

Bonding

Clean

Photo-resist

UV exposure

Patterning

Deep etching

Clean

-- Adhesively bonding (polymeric) methods --

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•  On-Chip magnetic manipulation

Flow

PCB

Microfluidic

Lee, H. et al, IEEE, Journal of Solid State Circuits, 2006.

CMOS Chip

Gasket

Glass

Manaresi, N. et al, IEEE, Journal of Solid State Circuits, 2003.

-- Rapid prototyping methods --

Microfluidic Packaging : Rapid prototyping!


Microfluidic Packaging : Direct-write assembly!

Robotically controlled

deposition (RCD) machine*

Cesarano, J., “Freeforming objects with low-binder slurry,” US patent. Therriault et al., Nature Materials, 2003.

•  Robotic based method for ink dispensing •  Ink : Sacrificial layer


•  Ink deposition –  Robotically controlled apparatus for

deposition in a predefined pattern;

–  Air-operated dispensing system for ink extrusion through micronozzle

–  Fugitive ink is binary organic ink composed of petroleum jelly and microcrystalline wax.

Micronozzle

Resin infiltration

Solidification of structural material

Removal of fugitive ink


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Removal of fugitive ink

Ink deposition

Before degassing

After degassing

During infiltration

Optical microscope images during resin infiltration process


•  Resin infiltration between patterned features •  Solidification (Resin cures at room temperature) •  Removal of fugitive ink under vacuum.

Femlab


(a)

(b)

•  Sensing electrodes

•  Fugitive Ink and fluidic Connection

•  Fugitive dam, epoxy resin, ink removel, and analyte injection.

(c)

Microfluidic Packaging : Direct-write assembly


•  Fluidic connections –  Between millimeter-scale fittings to micro-scale channel

•  Fixed tubing, tapered opening for syringe tip insertion

Common techniques employed for connecting / fittings to microchannels

Microfluidic Packaging : Direct-write assembly

Documents

Laboratory-on-Chip : Purpose · Laboratory-on-Chip : Purpose • A multidisciplinary approach to miniaturize biological assays or procedures in ... EZ Smart Monitoring Laboratory-on-Chip