Saliterman, Fundamentals of BioMEMS and Medical Microdevices, Ch.

Micro-Total-Analysis-
Systems
Introduction to BioMEMS
MN-BIO4600 Lecture 9
2015
 Micro-Total-Analysis-Systems
A device that automates and includes all necessary steps for chemical
analysis of a specimen e.g. sampling, sample transport, filtration, dilution,
chemical reactions, separation and detection:
Faster analysis and response times
short diffusion distances, high surface to volume ratios
rapid thermocycling (small heat capacities)
Smaller sample requirements and carrier volumes
Less waste, lower reagent consumption.
Safer platform
chemical, radioactive or hazardous biological studies
Improved transport
electrokinetic effects and miniaturised pumps
Compactness of a single device system
Integration of channels, mixers, separators, reaction chambers, electrodes and
detectors.
massive parallelization, permit high-throughput analysis
Lower fabrication costs
cost-effective disposable chips, fabricated in mass production
Microanalysis
 Task:
Designyour own total
analysis system
Design Opportunities
 Reaction Integrated
Valves Filter channel Detector/electrodes

Waste
Chip (outlet)

Intercon-
nectivity
(optional)

Actuators Reservoirs Pad ring Wash

(inlet) - electrical connections (buffer)
- power, user interface

Hybrids of multiple chips

 Chain of Operations

Fig 1: Lab-on-a-chip device.

 Applications
Real-time PCR
DNA amplification to detect bacteria, viruses and cancers

Biochemical assays
Cytotoxicity, DNA, Protein, RNA, Cell counting, viability, proliferation,
cytotoxicity, cellular secretions, virology and drugs

Immunoassay
Detect bacteria, viruses and cancers based on antigen-antibody reactions

Dielectrophoresis
Selection and trapping of particles, detection of cancer cells and bacteria

Blood sample preparation

Lysis of cells to extract DNA

Single-cell analysis
Ion channel screening (patch clamp)
Fully integrated BioChip
60 mm

100 mm
a b

Fig 14: Liu, RH, Analytical Chemistry, 76(7), 2004

Components
 Micromixer
Passivemicromixers:
Noexternalenergy
mixingreliesondiffusionor
chaoticadvection.
mixedphasesarranged:
parallellamination,serial
lamination,injection,chaotic
advectionanddroplet.
robust,andeasilyintegratedina
morecomplexsystem

Activemicromixers:
Useexternalenergyformixing
pressure,temperature,
electrohydrodynamics,
dielectrophoretics,electrokinetics
andacoustics.
Fig 2: Example of a mixer chip.
Paralell lamination micromixers

Fig 3: (a) basic T-mixer and (b) Y-mixer, (c) the concept of
parallel lamination and (d ) the concept of hydraulic focusing.
 Chaotic advection Mixers, Re > 200

Fig 4: (a) obstacles on wall, (b) obstacles in the channel and (c) zig-zag-shaped channel.
 Chaotic advection Mixers, Re < 20

Fig 5: Modification of mixing channel for chaotic advection at low Reynolds numbers:
(a) slanted ribs, (b) slanted grooves, (c) staggered-herringbone grooves and (d)( f )
patterns for surface modification in a micromixer with electrokinetic flows.
Chaotic advection Mixers, Re < 20

Fig 6: Laminar flow over staggered-herringbone grooves.

 Serial lamination micromixers

Fig 7: (a) joinsplitjoin, (b) splitjoin, (c) splitsplitjoin and (d ) multiple intersecting microchannels.
 Active micromixers

Fig 8: (a) serial segmentation, (b) pressure disturbance along the mixing channel, (c) integrated microstirrer in the
mixing channel, (d ) electrohydrodynamic disturbance, (e) dielectrophoretic disturbance, ( f ) electrokinetic
disturbance in the mixing chamber and (g) electrokinetic disturbance in the mixing channel.
Microchemical Reactors

Takesadvantageoflaminar
flowinmicrochannels
Transportbydiffusion
Welldefinedcontacttimes
Passiveoractivemixers
Immisciblephasesbrought
togetherandseparated
withoutphasemixing.
Singleormultiphase(liq/liq
andliq/gas)
Scalingpotential arrays

Fig 9: Examples of chemical reactor chips.

Principle of two-phase microcontactor

Fig 10: Microreactor

 Detection
Detectionstrategies:
Electrodesfor
electrochemicaldetection:
Metallayersmustbe
incorporated
Platinumisusuallyused
(alsogold,palladium)
Fluorescence:
Considerthewavelength
ofemission
below400nmrequire
fusedsilica(quartz)
Laser:
Refractionoflightisa
commondetection
method.

Fig 11: Examples of integrated electrodes.

 Interconnectivity

Interconnection
specificationsinclude:

Numberofports
Pressureandtemperature
operatingrange
Material(PEEKandstainless
steelareused)
Sizeofchipcapillaries
Methodofdetection(eg.
Microscopeaccess)

Fig 12: Examples of chip holders.

 Interconnect: Pressure Driven Flow

Fig 13: Analytical chip attached to a carrier with threaded interconnections

 Chip = 1 L
Dead Volume 10,000 L

Fig 15: Dead volumes in Lab-on-a-Chip applications

Fig 14: Analytical chip attached to a glass carrier with zero dead volume interconnectors
 Flow rate vs. applied pressure
1.0E+06

8QL d = 50 m
1.0E+04 P
R 4
1.0E+02
100 bar
Pressure / bar

1.0E+00
d = 1 mm

1.0E-02

1.0E-04 1 mbar

1.0E-06
Q

1.0E-08

1.0E-10
1.0E-16 1.0E-14 1.0E-12 1.0E-10 1.0E-08 1.0E-06
1 L

Flow / m3 s-1

Fig 15: Limitation in the delivery rate of feeding tubes

Electro-osmotic
Interconnect: flow
Electro-osmotic flow

100 V cm-1

Units in mm

Fig 16: Flow generated by the movement of charged

ionic compounds in an electric field
Assay Techniques
 Assay Techniques

Capillary array electrophoresis:

Linear and radial arrays

Cell, molecule and particle handling:

Mechanical, semipermeable membranes, electrokinetic,
chemical, optical, hydrogels

Surface modification:
Glass, silicon and polymers,
Immobilization strategies

Microspheres:
Bioseparations, latex agglutination test, drug delivery
Capillary Electrophoresis
Capillary Electrophoresis
Separation of ionic species based on
their size to charge ratio
(hydrodynamic radius, friction)
Utilise the interior of a small capillary
filled with an electrolyte
Sample introduced into capillary
capillary action, pressure, siphoning

Migration of the analytes is then

initiated by an electric field
Sample detection
UV absorption, fluorescence,

Fig 17: Separation by capillary electrophoresis.

 Sample detection

Fig 18: Optical detection by optical fibre.

 Optical path lenght

Fig 19: Techniques for increasing the pathlength of the capillary.

Microchannel flow profile

Fig 20: Flow profile laminar and electroosmotic flow.

 Modes of separation
ve eE ve = migration velocity (m/s)
e = electrophoretic mobility (m2/Vs)
(proportional to ionic charge of sample,
z inverse proportional to frictional forces)
e E = electric field strength (V/m)

6r z = net charge of analyte/particle

(eta) = viscosity (Pas) or (kg/sm)
r = stokes radius of analyte/particle (m)
k BT kB = Boltzmanns constant (1.38x10-23 J/K)
r T = temperature (K)

6D D = diffusion coefficient (m2/s)

L = distance inlet to detection point (m)
Lt = total length of capillary (m)
L L t tr = time required of the analyte to reach the
e detection point (s)

t r V
V = applied electric potential (V)

Fig 21: Modes of separation.

 Linear to radial arrays

Fig 22: Evolution from linear to radial capillary array electrophoresis.

 Radial capillary array electrophoresis

Fig 23: Combining parallell screening and extended separation channel length in a radial
configuration with a common anode.
Cell,Molecularand
ParticleHandling
Cell, Molecular and Particle Handling

Mechanical

Impedance Counting and Sizing

Semipermeable Membranes

Dielectrophoresis

Protein Adhesive Rolling

Optical Tweezers and Scissors

Hydrogels
 Mechanical separation

Separation by mechanical rather

than chemical means
Selective barrier
screen or filter cloth

Fluidic phase density

hydrostatic separators

Fluid and particle mechanics

Density, size, shape

Surface or electrical
characteristics of particles.
electrophoresis

Fig 24: Separation by filtering (silicon filter, 3m wide 50 m high pillars, 2m separation).
 Impedance counting and sizing

Impedance measurement as cells pass through silicon apertures

Displacement of electrolyte change in impedance over insulating wall
Constant current over electrodes change in voltage
Pulse shaped response from each cell (magnitude related to volume)
Thousands of cells analysed in seconds

Fig 25: The principle of impedance cell sizing.

Fig 26: Cell counter and separator, Sony Corp. 2011.
 Semipermeable Membranes
Permselective membranes for cell
immunoisolation:
High density uniform pores allow sufficient
permeability to nutrients and hormones
while preventing the passage of
immunoglobulins.

For example islet-cell transplantation

Uniform pores can be micromachining in

silicon

Polyethylene terephthalate (PET)

membranes may be machined with an
excimer laser to produce pores as
sieves

Fig 27: Silicon membrane array for immunoisolation and separation of biological components.
 Dielectrophoresis

Dielectric particles
Spatially nonuniform electric
field
experience a net force directed
toward locations with increasing
or decreasing field intensity
According to physical properties
of the particles and medium.

Potential uses include:

Isolation and detection of cancer
cells.
Concentration, separation,
trapping and positioning of cells
in dilute suspensions.

Fig 28: Separation of cells by dielectrophoresis

(Korea Advanced Institute of Science and Technology).
Protein adhesive rolling

Carbohydrate receptors on the

cell (leukocyte) wall
bind to (ligand proteins (e.g.
selectin) on the surface of a
vessel, with marginal affinity
This causes the leukocytes to
slow down and begin rolling
along the surface
Can be utilized to trap and sort
specific cells

Fig 29: Cell rolling along a surface.

 Optical Tweezers and Scissors

Tweezers:
Moving particles with light
Transfer of momentum from a
focused laser to a particle.
Particle trapped near beam focus.
Pushed axially in direction of beam.
Gaussian beam:
Trapping of particles in axial
direction limited to few
micrometers apart.
Bessel beams:
does not diverge + reconstructs
after obstruction
Multiple trapping of cells

Scissors:
Lysis of cells

Fig 30: Principle of Operation.

 Groves in the Zona pellucida

Fig 31: Application of optical tweezers/scissors.

 Hydrogels

Control of microfluidic flow

operation:
Swelling from chemical stimulus
Temperature
Control by secondary flow
stream (isolation of hydrogel
from main stream accomplished
by membrane).

Fig 32: Microfluidic hydrogel valve that swell

by chemical stimulus and closing the valve.
Fig 33: Temperature sensitive tethered hydrogel valves fabricated into a silicon membrane.
Fig 34: Isolation of the hydrogel from the main stream
by thin film membrane.
Surface Modifications
 Surface modifications

Chemical surface modification

enhance electrokinetic effects and create areas of
hydrophobicity, hydrophilicity and adhesion that assist in
fluid handling
Biological surface science
the study of properties and processes at interfaces between
synthetic materials and biological environments and
fabrication of biofunctional surfaces
Protein or cell-resistant surfaces
a requirement for medical devices that are in contact with
biological fluids
Immobilization strategies
Control of fluids and positioning of analytes for reactions and
detection
 Glass and silicon substrates

Silicon dioxide surfaces (and glass) are negatively

charged at neutral pH due to deprotonated silanol
groups, and are hydrophilic
An EDL will form when in contact with an electrolyte
solution.

To control EOF, both the sign of the surface charge and its
distribution on the surface are important

Discrete areas of positive charge can be achieved by

patterning with positively charged poly(allylamine
hydrochloride) (PAH)

Electro-deposited silver may increase hydrophobicity

allowing control of flow.
 Polymer substrates
(selection dependent on following properties)

Wide range of substrates with different (hydrophobic)

surface chemistries
Machinability

Solvent resistance

Hydrophobicity (PMMA)

Zeta-potential and associated electroosmosis flow

mobility (ratio of flow rate to electric field)
Surface bond chemical moieties

Non-specific adsorption of analytes

 Techniques

Covalent chemical modification:

Surfaces retain chemical integrity over an extended period
of time compared to non-covalent modification
Reactions with pendent groups typically occur

Energetic surface treatments:

UV and plasma (Ar, Ne, He, H2, NH2, CO, CO2, O2,H2O, N2,
NO2 and F2)
May enhance hydrophilicity, hydrophobicity, adhesion,
surface charge density, biocompatibility and permeability

Non-Covalent Modification
Protein and other surfactant coatings
 PEG and Gold Surface Modification

Fig 35: Surface modification for (a) silicon substrate with PEG, and (b) gold patterned silicon
substrate for guided cell adhesion.
 Immobilization Strategies

Mechanical barriers and traps

Laser tweezers

Surface modification:
Hydrophilicy and hydrophobicity as discussed.
Lipid bilayers offer efficient reduction of nonspecific cell
and protein binding.
Biospecific reactions such as with biotin/streptavidin.
Step modification of a substrate surface by first
immobilizing proteins, peptides, or carbohydrates as a
means to promote cell attachment.

Hydrogel matrices
 Summary

Biomedical Micro Electro-Mechanical Systems

Topics of study (curriculum):
Introduction to BioMEMS
Principles of Biochemistry
Silicon and Soft Fabrication Techniques
Polymer Materials
Microfluidic Principles
Sensor Principles and Microsensors
Microactuators and Drug Delivery
Clinical Laboratory Medicine
Micro-Total-Analysis Systems
Detection and Measurement Methods
Genomics and DNA Microarrays
Proteomics and Protein Microarrays
Emerging BioMEMS technologies
Packaging, Power, Data, and RF Safety
Biocompatibility, FDA and ISO 10993
Thank you