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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.
GBM8320 - Dispositifs Médicaux Intelligents 3
• Low sample consumption;
• Fast biochemical reactions & Miniaturization;
• Point-of-care.
Laboratory-on-Chip : Advantage
2
GBM8320 - Dispositifs Médicaux Intelligents 4
Laboratory-on-Chip : CMOS technology
• Low voltage, and low power;
• Addressable sensing or manipulating sites;
• Standard technology.
GBM8320 - Dispositifs Médicaux Intelligents 5
I. Introduction
II. Biochemistry
III. Microfluidic Packaging
IV. Capacitive Sensors
V. On-Chip Cells Detection and Manipulation.
Laboratory-on-Chip : Outline
GBM8320 - Dispositifs Médicaux Intelligents 6
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!
3
GBM8320 - Dispositifs Médicaux Intelligents 7
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
GBM8320 - Dispositifs Médicaux Intelligents 8
Affymetrix Inc.!
Requires scanner for reading out!
EZ Smart Monitoring
Laboratory-on-Chip : Technologies
Glucose Detection DNA !array Rapid Bacterial Diagnosis
STMicro-electronics
GBM8320 - Dispositifs Médicaux Intelligents 9
Laboratory-on-Chip : Future Technologies
Display
Disposable needle
Sensor
Syringe Microchannel
Past Room
Laboratory
Today Desktop
Laboratory
Future Pencil-Size Laboratory
4
GBM8320 - Dispositifs Médicaux Intelligents 10
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
GBM8320 - Dispositifs Médicaux Intelligents 11
• 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.
GBM8320 - Dispositifs Médicaux Intelligents 12
,...,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!
5
GBM8320 - Dispositifs Médicaux Intelligents 13
! 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!
GBM8320 - Dispositifs Médicaux Intelligents 14
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!
GBM8320 - Dispositifs Médicaux Intelligents 15
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.
6
GBM8320 - Dispositifs Médicaux Intelligents 16
I. Introduction
II. Biochemistry
III. Microfluidic Packaging
IV. Capacitive Sensors
V. On-Chip Cells Detection and Manipulation.
Laboratory-on-Chip : Outline
GBM8320 - Dispositifs Médicaux Intelligents 17
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!
GBM8320 - Dispositifs Médicaux Intelligents 18
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.
Laboratory-on-Chip : Fundamental of Biochemistry!
7
GBM8320 - Dispositifs Médicaux Intelligents 19
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.
Laboratory-on-Chip : Fundamental of Biochemistry!
Polymerase Chain Reaction (PCR)
Centrifuge system Fragmentation
PCR
Cells
DNA
Molecules
GBM8320 - Dispositifs Médicaux Intelligents 20
Laboratory-on-Chip : Fundamental of Biochemistry!
• 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.
GBM8320 - Dispositifs Médicaux Intelligents 21
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).
8
GBM8320 - Dispositifs Médicaux Intelligents 22
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..
GBM8320 - Dispositifs Médicaux Intelligents 23
• 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
Laboratory-on-Chip : Fundamental of Biochemistry!
Antibody
Antigen
GBM8320 - Dispositifs Médicaux Intelligents 24
I. Introduction
II. Biochemistry
III. Microfluidic Packaging
IV. Capacitive Sensors
V. On-Chip Cells Detection and Manipulation.
Laboratory-on-Chip : Outline
9
GBM8320 - Dispositifs Médicaux Intelligents 25
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
GBM8320 - Dispositifs Médicaux Intelligents 26
• 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!
GBM8320 - Dispositifs Médicaux Intelligents 27
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 --
10
GBM8320 - Dispositifs Médicaux Intelligents 28
• 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!
GBM8320 - Dispositifs Médicaux Intelligents 29
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
GBM8320 - Dispositifs Médicaux Intelligents 30
• 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
Microfluidic Packaging : Direct-write assembly!
11
GBM8320 - Dispositifs Médicaux Intelligents 31
Removal of fugitive ink
Ink deposition
Before degassing
After degassing
During infiltration
Optical microscope images during resin infiltration process
Microfluidic Packaging : Direct-write assembly!
• Resin infiltration between patterned features • Solidification (Resin cures at room temperature) • Removal of fugitive ink under vacuum.
Femlab
GBM8320 - Dispositifs Médicaux Intelligents 32
(a)
(b)
• Sensing electrodes
• Fugitive Ink and fluidic Connection
• Fugitive dam, epoxy resin, ink removel, and analyte injection.
(c)
Microfluidic Packaging : Direct-write assembly
GBM8320 - Dispositifs Médicaux Intelligents 33
• 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