Nanotechnology in Biosensors

Nanowire Biosensors
Nanotube Biosensors
Nanoparticles
FET and CMOS devices
Nanotechnology (sometimes shortened to "nanotech") is the study of
manipulating matter on an atomic and molecular scale. Generally
nanotechnology deals with structures sized between 1 to 100 nanometer in at
least one dimension, and involves developing materials or devices within that
size. Quantum mechanical effects are very important at this scale.

Drexel University materials science graduate

student Jennifer S. Atchison made the silicon
nanocones shown in this scanning electron
microscope image by decomposing silane at
high temperature in a chemical vapor
deposition apparatus.
This transcript of the classic talk that Richard Feynman gave on December 29th 1959 at the
annual meeting of the American Physical Society at the California Institute of Technology
(Caltech) was first published in the February 1960 issue of Caltech's Engineering and
Science.

Miniaturizing the computer

I don't know how to do this on a small scale in a
practical way, but I do know that computing
machines are very large; they fill rooms. Why can't
we make them very small, make them of little wires,
little elements---and by little, I mean little. For
instance, the wires should be 10 or 100 atoms in
diameter, and the circuits should be a few thousand
Richard Feynman angstroms across.
 What is nanostructure?
One dimention 1-100 nm

2-D Structures (1-D confinement)

2 µm
• Thin Films
• Planar Quantum wells
• Superlattices Si Nanowire Array

1-D Structures (2-D confinement)

• Nanowires
• Quantum wires
• Nanorods
• Nanotubes

0-D Structures (3-D confinement)

• Nanoparticle
• Quantum Dots Multiwall Carbon
Nanotube
 Energy Levels in a Metal

Bulk Material Large Nanoparticle Small Nanoparticle

 Nanowires
• Solid, “1D”

• Conductive, Semi-conductive, Insulator

• Crystalline

• Quantum Confinement Effect (electon, phonon)

2 µm
• Decrease in nanowire diameter resulted Si Nanowire Array
with increase in band gap

• Thermal conductivity decreases with decreasing

nanowire diameter

• Core-shell synthesis is possible

Si/SiGe Nanowires
Publications

“Web of Science” September 2007 data; 4699 publications

December 2010 data; 13186 publications
Nanowire Synthesis
Nanowire Synthesis

• Electron Beam Lithography

• Molecular Beam Epitaxy (MBE)
• Thermal Evaporation
• Laser-Ablation
• Template Assisted Synthesis
High Pressure Injection
Vapor Depostion
Electrodeposition
Template Assisted Au catalyzed Si Nanowire Synthesis

Template Above

Au Nanoparticles

1µm

100nm
Chemical Vapor
Deposition(CVD)

Nat. Protocols 1, 1711-1724 (2006).

Si Nanowires

Basınç (Tor)
30-70

Sıcaklık Proses Gazı/Taşıyıcı Gas

75 25
480-500 ºC
50 50
Zaman 25 75
10 saniye-25 SiH4/N2

dakika
Au Kolloid Boyutu
2-20 nm

Au Kolloid Derişimi
0.01-1
 Nomenclature

• CVD - Chemical Vapor Deposition

• MWNT - Multi-Walled Carbon Nanotubes
• SWNT - Single-Walled Carbon Nanotubes
• XRD - X-ray Diffraction
• Pyrolysis - decomposition of organic material through the
application of heat and the absence of oxygen
• Chirality - measure of the twist of the nanotube
• Ablation - Removing a surface material by vaporization

15
 What Are Carbon Nanotubes?
• CNT can be described as a
sheet of graphite rolled into a
cylinder
• Constructed from hexagonal
rings of carbon
• Can have one layer or multiple
layers
• Can have caps at the ends
making them look like pills

Information retrieved from: http://www.photon.t.u-tokyo.ac.jp/~maruyama/agallery/agallery.html

16
 Nanotube Classification
• Chirality - twist of the
nanotube
• Described as the vector
R (n, m)
• Armchair vector, R vector,
angle
• = 0º,φarmchair nanotube
• φ0º < < 30º, chiral nanotube
• φ zigzag nanotube
> 30º,
φ

Information and image retrieved from: http://www.pa.msu.edu/cmp/csc/ntproperties/

17
 Nanotube Classification
• MWNT
– Consist of 2 or more layers of
carbon
– Tend to form unordered clumps
• SWNT
– Consist of just one layer of
carbon
– Greater tendency to align into
ordered bundles
– Used to test theory of nanotube
properties

Images retrieved from: http://www.photon.t.u-tokyo.ac.jp/~maruyama/agallery/agallery.html

18
 Applications
• Electronic Devices
– Nanotube TV’s
– Nano-wiring
• High Strength Composites
– 100 times as strong as steel and 1/6 the weight
• Conductive Composites
• Medical Applications
– Encase drug into nanotube capsule for more
predictable time release

19
 Synthesis Techniques
– Nanotube Synthesis By CVD Process

– Plasma Enhanced CVD Nanotube Synthesis

– Nanotube Synthesis By Arc Discharge in a Magnetic Field

– Carbon Nanotube Synthesis Using Laser Ablation of Metallic Catalyst

“Web of Science December 2010 data; 27117 publications

20
 Chemical Vapor Deposition
1. Gas enters chamber at room
temperature (cooler than the
reaction temperature)
2. Gas is heated as it approaches
the substrate
3. Gases then react with the
substrate or undergo chemical
reaction in the “Reaction Zone”
before reacting with the
substrate forming the deposited
material
4. Gaseous products are then
removed from the reaction
chamber

Information and photo retrieved from:

21 http://www.sandia.gov/1100/CVDwww/cvdinfo.htm
Nanotube Synthesis By CVD Process

Schematic from: Andrews, Jacques, Qian, and Rantell, “Mulitwall Carbon Nanotubes: Synthesis and Application”
22
 Nanotube Synthesis By CVD Process
• Source of carbon atoms usually comes from an
organic compound
• Mixed with a metal catalyst and inert gas
• Atomized and sprayed into reactor with
temperatures ranging from 600ºC to 1200ºC
• Pyrolysis of organic compound deposits
carbon (as soot) and carbon nanotubes on
reactor wall (usually a tube constructed from
quartz)
23
 Sources of Carbon
• Typical Organic/Catalyst Mixtures
– Xylene/ferrocene
– Toluene, benzene, xylene, mesitylene, and n-
hexane/ferrocene
– Ethylene and ethanol/Fe, Co, and Mo alloys

• Typical Carrier Gases

– Argon
– Hydrogen

24
 Plasma Enhanced CVD Nanotube
Synthesis
• Methane moves toward the
catalyst on the substrate
• Heat of the reactor
decomposes methane at the
catalyst surface
• Catalyst is at a slightly cooler
temperature so carbon is
supersaturated in the catalyst
film so carbon precipitates out
• Carbon forms nanotubes at
the surface of the catalyst film

25
Nanowire sensor approach
• Approach for the direct electrical detection of
biological macromolecules uses semiconducting
nanowires or carbon nanotubes configured as field-
effect transistors, which change conductance upon
binding of charged macromolecules to receptors
linked to the device surfaces

Carbon nanotubes on the electrode

Field effect transistor (FET).. An overview

• The name ‘transistor’ is a shortened version of the

original term, transfer resistor
• Relies on using one electrical signal to control
another
• The effect is to make a ‘resistance’ whose value can
be altered by the input signal.
• This behaviour can be used to ‘transfer’ patterns of
signal fluctuation from a small input signal to a
larger output signal.
Again… Semiconductors…
• Group III-V single-crystal semiconductors are used
for detection of a target analyte.
• If changes are reversible and detectable by means of
cahnges in optical and/or electrical properties, on-line
detection is possible.
• Adsorbate can be categorized as donors (D) or
acceptors (A) and can react in the following way:
• D
D++e-
• A+e-
A-
 Semiconductors

• For silicon and germanium, the Fermi level is essentially

halfway between the valence and conduction bands.
• No conduction occurs at 0 K, at higher temperatures a finite
number of electrons can reach the conduction band and provide
some current. In doped semiconductors, extra energy levels are
added.
Silicon Energy Bands

• At finite temperatures, the number of electrons which reach the conduction band
and contribute to current can be modeled by the Fermi function. That current is
small compared to that in doped semiconductors under the same conditions.

• The increase in conductivity with temperature can be modeled in terms of the

Fermi function, which allows one to calculate the population of the conduction
band.
P-Type Semiconductor

Lack of electrons results in

lowering fermi level down to
valance band

• The addition of trivalent impurities such as boron,

aluminum or gallium to an intrinsic semiconductor creates
deficiencies of valence electrons, called "holes".
N-Type Semiconductor

Fermi level rises up to

conduction band in
negative type semi conductor

The addition of pentavalent impurities such as antimony, arsenic

or phosphorous contributes free electrons, greatly increasing the
conductivity of the intrinsic semiconductor.
Bands for Doped Semiconductors

• In n-type material there are electron energy levels near the top of
the band gap so that they can be easily excited into the
conduction band.
• In p-type material, extra holes in the band gap allow excitation
of valence band electrons, leaving mobile holes in the valence
band.
 Semiconductor Current
• The electrons which have been freed from their lattice positions

into the conduction band can move through the material.

• Holes are migrating across the material in the direction opposite

to the free electron movement.

• This current is highly temperature dependent.

•The current flow in a semiconductor is influenced by the density of energy

states which in turn influences the electron density in the conduction band.
P-N Junction – coming closer to transistors…

• When p-type and n-type materials are placed in contact with each other, the junction
behaves very differently than either type of material alone.

• Specifically, current will flow readily in one direction (forward biased) but not in the other
(reverse biased), creating the basic diode.

• The open circles on the left side of the junction above represent "holes" or deficiencies of
electrons in the lattice which can act like positive charge carriers. The solid circles on the
right of the junction represent the available electrons from the n-type dopant.

• Near the junction, electrons diffuse across to combine with holes, creating a "depletion
region".
Depletion Region

• When a p-n junction is formed, some of the free electrons in the n-region
diffuse across the junction and combine with holes to form negative ions.
In so doing they leave behind positive ions at the donor impurity sites.
Depletion Region Details
• In the p-type region there are holes from the acceptor
impurities and in the n-type region there are extra
electrons.

• When a p-n junction is formed, some of the electrons from

the n-region which have reached the conduction band are
free to diffuse across the junction and combine with holes.

• Filling a hole makes a negative ion and leaves behind a

positive ion on the n-side. A space charge builds up,
creating a depletion region which inhibits any further
electron transfer unless it is helped by putting a forward
bias on the junction.
 Forward bias and Reverse bias
• An applied voltage in the forward direction as
indicated assists electrons in overcoming the
coulomb barrier of the space charge in depletion
region. Electrons will flow with very small
resistance in the forward direction.

• An applied voltage with the indicated polarity further

impedes the flow of electrons across the junction. For
conduction in the device, electrons from the N region
must move to the junction and combine with holes in
the P region. A reverse voltage drives the electrons
away from the junction, preventing conduction.
Forward Biased Conduction

• The forward current in a p-n

junction (forward-biased) involves
electrons from the n-type material
moving leftward across the
junction and combining with holes
in the p-type material.
Field effect transistor
•A pair of metallic contacts are
placed at each end of the channel.
When we apply a voltage between
these, a current can flow along the
channel from one contact to the
other. The contact which launches
charges along the channel is called
the source, the one that 'eats' them
at the other end is called the drain.

•In a sense, the device is a bit like

a PN-junction diode, except that
we've connected two wires to the
N-type side.
 Forward bias

p- or n
n-- type silicon
nanowire (SiNW)
as semiconductor

Receptors acting as
acceptor or donor
on semiconductor yielding
Field Effect Transistor
arrangement
FET amplifies the result
(conductivity change)
..A nanowire arrangement
1D SiNWs as field effect transistors
• Binding to the surface of a nanowire can
lead to depletion or accumulation of charge
carriers in the “bulk” of the 1D structure
(over nanowire).

• Remember that in planar FET, charge

defficiency was only on the surface region
of the device.

• This increase sensitivity (single molecule

detection possible)
Sensitivity
• Binding strength dependent on Fermi level
(equilibrium binding constant)
• Binding strength
Analyte detection sensitivity
(detection limit)
• Fermi level
n or p-type semiconductor (crystal
growth, impurities)
• Amplification on FET structure
binding properties
• Sensitivity increment
irreversibility on binding
Si Nanowire Field Effect Transistors (FET)

Nat. Protocols 1, 1711-1724 (2006).

Si Nanowire Field Effect Transistors (FET)

Nat. Protocols 1, 1711-1724 (2006).

Si Nanowire Field Effect Transistors (FET)

Nat. Protocols 1, 1711-1724 (2006).

Si Nanowire Field Effect Transistors (FET)

Virus Detection

Influenza A

Adenovirus

80 Attomolar (10-18)

Appx. 50 Virus/mL

Proc. Natl. Acad. Sci. USA 101, 14017-14022 (2004).

Si Nanowire Field Effect Transistors (FET)

DNA SENSORS

60 Femtomolar PNA

100 HeLA Cells

Nat. Biotechnol. 23, 1294-1301 (2005).

Si Nanowire Field Effect Transistors (FET)

Prostat Spesific Antigen

2 Femtomolar

PSA
Carsinoembriyonic Antigen
Musin 1

Nat. Biotechnol. 23, 1294-1301 (2005).

Carbon Nanotubes
Carbon Nanotubes
Carbon Nanotubes
Carbon Nanotubes
Carbon Nanotubes
Carbon Nanotubes
 NANOPARTICLES

Metalic Nanoparticles
(Au, Ag)

Polymeric Nanoparticles

Quantum Dots
Quantum Dots
3 nm
 Quantum Dots
• Quantum dots are very monodisperse inorganic nanocrystalline particles made
from semiconducting materials which are crystals composed of periodic groups of
II-VI, III-V, or IV-VI materials.

II-VI :CdSe, CdTe, CdS, and ZnSe

III-V :InP and InAs
IV-VI :PbS, PbSe

• These particles are typically 2-10 nm (10-50 atoms) in size, about the size of
typical proteins.

• QDs are also referred to “artificial atoms”

Quantum dots are nanometer-scale crystals that were developed in the mid-
1980s for optoelectronic applications.
 Quantum Dots
The most intriguing features of QDs is that the particle size determines many of the QD
properties most importantly the wavelength of fluorescence emission.

Colloidal CdSe quantum dots dispersed in hexane

Individual quantum dots are too small to see

with the naked eye, but they signal their
presence by emitting light in a variety of
colors. They emit different colors depending
on their size.
 Quantum Dot Type
Core type Qdot Core/shell type Qdot

c
Core-
shell
coating

Core
Quantum
Dot

CdSe Core Qdot CdSe/Zns Core/shell Qdot

Cd S

Se Zn
Semiconductor nanocrystals can also be produced with other shapes such as;

 Pyramids  Boomerangs
 Rods  Tetrapods, or many other shapes
 Spheres Biological application
 Quantum Dots Structure
Biomolecules(SA)
Polymer coating
Inorganic shell (ZnS)

Nanocrystal
Core (CdSe)

Core Shell
The first step in synthesizing Qdot  The shell layer composed of a semicoductor
nanocrystals is the preparation of a core that material of wider bandgap than the core
is composed of : material.
 Cadmium sulfide (CdS): UV-blue
 Cadmium selenide (CdSe): the bulk of  Stabilize the material,
the visible spectrum
 Improve quantum yield (increases the
 Cadmium telluride (CdTe): the far red
intensity of the fluorescence)
and near-infrared
The core nanocrystal determines the
 Reduce photo-degradation.
color
 Optical Properties
Color Tunability

Tuning the QD emission wavelenght by changing the particle size or composition

QDots are available in multiple emissions from 465 nm (visible) to 2300 nm (infared)

Quantum Dot size Quantum Dot Composition

A
 Optical Properties
Narrow Fluorescent Emission

Qdots exhibit narrow and symmetric emission peaks (FWHM typically 25-35 nm).

Broadband Absorption Source

QDots have a broader absorption profile compared to traditional organic dyes and phosphors.

CdSe Quantum Dot FITC: Fluorescein isothiocyanate

(organic dye)

Wavelength (nm) Wavelength (nm)

Simultaneous labeling and detection of multiple analytes

 Optical Properties
Em
issi
on
Wa
vel

Emission Wavelength (nm)

Excitation 360 nm

All samples are induced to emit their respective

Vials of Quantum Dots in front of a common UV colors even though a single source was used to
hand lamp excite them.

Different size QDs can be excited with a simple excitation sources

This facilitates simultaneous detection, imaging and quantification

 Optical Properties
Stability
The nanocrystal materials are very stable. They are composed of inert inorganic compounds
and are further stabilized with our engineered shells that resist photochemical damage.

Intensity
QDot nanocrystals fluoresce intensely and exhibit high quantum yields. Brightness is
comparable to or greater than traditional organic fluore dyes.

organic-dye

The signal is seen to

fade in time

quantum dots
The same time
fluoresce brightly for
much longer

In vivo imaging of Xenopus embryos using time-lapse

microscopy
 Optical Properties
Fluorescence Lifetimes
CdSe/ZnS nanocrystals have 15-20ns fluorescence lifetimes which is an order of magnitude
greater than conventional organic dyes and even greater than the auto-fluorescence lifetime of
organic dyes.

A: Nuclear antigen: QDot (red) B: Nuclear antigen: Alexa 488 (Green)

Microtubules: Alexa 488 (Green) Microtubules: QDot (red)

 Synthesis of Quantum Dots

Vapor Deposition

Ion Implantation

Sol-gel Method

Micelle Method

Organometallic Synthesis via Pyrolysis

Organometallic Synthesis via Pyrolysis
Core Type Qdot (CdSe)

TOP: Trioctylphosphine TOPO: Trioctylphosphine oxide

HPA: Hexylphosphonic acid

Precursor: CdO
 Organometallic Synthesis via Pyrolysis
High-temperature Pyrolytic Reaction
Core/shell Type Qdot (CdSe/ZnS)

Metallic or organometallic precursor Zinc, Chalcogen precursor

cadmium or mercury species Sulfur, selenium or tellurium species

Coordinating solvent (TOPO,TOP, hexadecylamine)

High temperatures
Organometallic Synthesis via Pyrolysis
QDs obtained from these procedures are:

 Highly fluorescent,
 Photostable,
 Sufficiently monodisperse for use as labels in biological studies,
 But they are not soluble in aqueous solution and biocompatible

Surface modification Bioconjugation

 Surface Modification
Two general methods
LIGAND EXCHANGE
A:Surface -exchange of hydrophobic
surfactant molecules for bifunctional
linker molecules

TOPO-coated QD replaced with

mercaptoacetic acid or silane

B:Phase-transfer methods using A

amphiphilic molecules that act as detergent HYDROPHOBIC INTERACTIONS
for solubilizing the QD coated with
hydrophobic groups.

TOPO ligands on the surface may be

interact with an amphiphilic polymer
(octylamine-modified polyacrylic acid). B
 Bioconjugation Methods
a) Bifunctional linkage d) Electrostatic attraction

b) Hydrophobic attraction

e) Nanobeads
c) Silanization

Two to five protein molecules

+ conjugated to a single 4 nm QD
50 or more small molecules
(oligonucleotides or peptides)
Biological Applications
 Qdots Based Array
Barcoding technology

Fluorescence Intensity
Code readout

The use of 6 colors and 10 intensity levels can encode 1

million protein or nucleic acid sequences

Wavelength

“Barcoding” technology can be used for

gene profiling and high-throughput drug and
disease screening

Polystyrene beads are embedded with multicolor Nature Biotechnology 19:631-635-97 (2001), ‘Quantum-dot-tagged microbeads
CdSe/ZnS QDs for multiplexed optical coding of biomolecules’
 Qdots Based Array
Prepared multicolor QD-tagged beads, and conjugated these beads to biomolecules
to carry out biological assays

Designed a model DNA

hybridization system using
oligonucleotide probes and
triple--color encoded beads
triple

DNA hybridization assays using QD-

tagged beads. Probe oligos (No. 1–4) were
conjugated to the beads by crosslinking,
and target oligos (No. 1–4) were detected
with a blue fluorescent dye such as
Cascade Blue. After hybridization,
nonspecific molecules and excess reagents
were removed by washing
 Qdots Based Array
Encoding

Application Format

Multiplex gene Encoded beads each with

expression gene-spesific oligo

Multiplex SNP Encoded beads with gene-

genotyping spesific oligos

Multiplexed Encoded beads with

immunoassays antibodies

Multiplexed cell- Mixed cell yypes encoded

based assays with qdots
dot
Quantum dots Spectrally-encoded
colors
colors mixtures Encoded cell lines
Multiplexed reporter
Encoded beads, cells, etc. assays expressing various
receptors

Mix to form
unique
mixtures Polymer beads (oligos), various
(codes) cell types, or cells with diferrent
receptors
 Qdots Based Array
Biological Threat Detection System Using QDs

Specific target
UV Light
Excitation

QD Nanoprobes Nanoassembly Colocalization

Colocalization

NANOLETTERS 2005,Vol. 5, No. 9,1693-1697, “Multiplexed Hybridization Detection with Multicolor Colocalization of Quantum Dot Nanoprobes”.
 Qdots Based Array
Simulated multiplexed analysis of anthrax-related genetic targets for pathogenicity

C I II
A

III IV

B: Four samples containing different combinations of the three

A: Color pallet for the three pairs of target specific QD
targets rpoB, capC, and pagA. Checks represent the existence of
nanoprobes and their resulting colocalized fluorescent
certain target sequences. Sample IV does not contain any target and
images upon sandwich hybridization.
is used as a negative control.

C: Fluorescent images I, II, III, and IV correlate with samples I, II, III, and IV,respectively.
NANOLETTERS 2005,Vol. 5, No. 9,1693-1697, “Multiplexed Hybridization Detection with Multicolor Colocalization of Quantum Dot Nanoprobes”.
Metalic Nanoparticles
 Metalic Nanoparticles
• Metalic nanoparticles possess optical properties that make them uniquely suitable for
biosensing applications.
• Their optical properties strongly depend on both the particle size and shape and are related to
the interaction between the metal conduction electrons and the electric field component of the
incident electromagnetic radiation, which leads to strong, characteristic absorption bands in
the visible to infrared part of the spectrum.
• In aqueous solutions, gold nanostructures exhibit strong plasmon bands depending on their
geometric shape and size.

Gold (Au)

• Gold nanoparticles are particularly attractive for studies in a biological environment

because they show no surface oxidation
• High biocompatibility without any surface modification.
• In addition, thiol chemistry can be applied to conjugate molecules to the gold surface.

LSPR (Localized Yüzey Plasmon Resonans) nanosensors can be used as diagnostic

tools for a variety of diseases
Size and Geometry Variation on Surface Plasmon
Absorption

• Size change in Au spheres

has little effect on absorption
maximum position

• Au nanorods have two

absorption peaks. One is due to
plasmon setup along the
axial direction and the other
one is along the radial
direction

• The longitudinal plasmon

absorption position changes with
aspect ratio.
 Colloidal form of Gold –Silver
particles for different shapes and
sizes

Au-Ag alloy
nanoparticles

Au nanorods

Ag nanoprisms
 Optic Properties of Au and Ag Nanoparticles

Au nanospheres Ag nanoparticles

Ag nanoparticles Ag nanoparticles

Ag nanoparticles Ag nanoparticles
 SPR (Surface Plasmon Resonance)
Modification of sensor surface
Au film

Probe immobilization
Glass slide

Injection of analyte solution

Binding

Changing of refraktive index or

dielectric constant
LSPR (Localized Surface Plasmon Resonance)

The nanoparticles have tunable optical

properties, which make them an ideal LSPR-
sensing platform. By functionalizing the
nanoparticle surface with the appropriate
receptor, the LSPR nanosensor can be used to
detect specific ligands
Silanization of glass İmmobilization of Au +
surface nanoparticles on the glass
surface
Sensing surface is exposed to the target
molecules

The refractive index of the surrounding

enviroment changes on binding to the
receptors, inducing a shift in the
nanoparticles LSPR,λmax . The
wavelength shift is monitored by UV-
Visible spectrocopy .
 LSPR (Localized Surface Plasmon Resonance)

Nanosphere Lithography (NL)

 LSPR Sandwich Assay
Anti ADDL antibody

ADDL antibody

Ag nanoparticles after modification

with anti- ADDL antibody

after exposure to ADDL antibody

And anti -ADDL antibody

Biomarker for Alzheimer’s disease

 Synthesis of Gold Nanoparticles
Spherical gold nanopartciles are easily produced by the chemical reduction of gold salts

Chlorauric acid
solution (HAuCl4) Boil 15 min

Tri sodium citrate or

NaBH4 solution

2.5×10-4 M chlorauric acid solution

• Bring to a boil 50 mL of 2.5×
• Add 0.16 to 1.0 mL of 34 mM sodium citrate solution to the boiling solution while stirring
• After a minute will be faint blue and then darkening over 5 min to a brilliant red
 Synthesis of Au Nanorods
Seed-mediated growth procedure

I. Seeds are prepared by

reducing of a metal salt
with a strong reducing
agent (Na-citrate or
NaBH4)

II. The growth steps

involve the addition of
more metal salt to the
seed solution, with a
weak reducing agent, in
the presence of a
surfactant

Capping agent: CTAB

Weak reducing agents:
ascorbic acid

The nanorod growth reaction was terminated after 3 h by removing the reaction solution by centrifugation at
5000 rpm for 15 min. The nanorods were resuspensed in 0.005 M CTAB solutions and found to remain stable for
up to 100 days.
 Bioconjugation Methods
Thiol end groups (-SH) are enough to covalently bond the molecules to the gold surfaces
surfaces.. It is
quite straight forward, easy and quick reaction, and requires no other chemicals.
chemicals. It is possible to
couple molecules carrying thiol end group on gold surfaces simply

Au Nanoprobes
+
SH
SH

SH

SH
SH

(Mercaptoundeconic acid
(MUA))
Probe Molecules

• Oligonucleotides
• Oligopeptides/proteins
• Antibodies

The alkyl chains can be terminated by reactive head groups, such as carboxylic acid (COOH) and
amino groups (NH2), are used for formation of funtionalized SAMs.
SAMs. These alkyl chanis can be
covalently bond to gold surfaces to act as spacer arm for further immobilization steps
steps.. These
sublayers are capable of supporting the immobilization of biomolecules via covalent chemical
coupling..
coupling
Multiplexing Ag Nanosensor Carbohydrate-
Carbohydrate-Sensing
Chip
Ag nanobiosensor consisted of nanosphere
litography-fabricated Ag nanoparticles with
the same in –plane width but two different out-
of-plane heights: 35 and 75 nm and thus two
different LSPR λmax.
724.5 nm
682 nm (B) Ag nanoparticles (75 nm height) mannose
677 nm
724.6 nm modification
(C) Ag nanoparticles (75 nm height) after
exposure to Con A.
(D) Ag nanoparticles (35 nm height) galactose
modification
(C) Ag nanoparticles (35 nm height) after
exposure to Con A.

Mannose

Galactose

Concavalin A
 Multiplex Biosensor Using Gold Nanorods

Gold nanorod molecular probe

Gold nanorods (GNRs) with different aspect ratios

were fabricated through seed-mediated growth and
surface activation by alkanethiols for the attachment of
antibodies to yield gold nanorod molecular probes
(GNrMPs). Multiplex
sensing was demonstrated by the distinct response of
the plasmon spectra of the GNrMPs to binding events
of three targets (goat anti-human IgG1 Fab, rabbit
antimouse IgG1 Fab, rabbit anti-sheep IgG (H+L))
 Multiplex Biosensor Using Gold Nanorods
(a) (b)

(c)

Multiplexing detection of various targets using GNrMPs

(a) One target; (b) two targets; (c) three targets
 Solution-phase Nanoparticles

2. Releasing the Ag nanoparticles into solution.

3.Asymmetrically linked solution-phase Ag

nanoparticles with alkanedithiol to form dimer.

Surface-bound Ag nanoparticles fabricated by nanosphere

lithograpy and modified with a SAM of alkanethiol.
Market For Biosensors
Need for Biosensor
Diagnostic Market. Diagnostics represent a very large and well established
market that is continually expanding. Particularly in the current climate of
prevention rather than remedy, the need for detection at increasingly lower
limits is increasing in many diverse areas. Estimates of the market size and
future projections tend to be difficult and inaccurate.
Need for Biosensor

Clinical Testing. However, undoubtedly clinical testing iis one of the biggest
diagnostic markets. A study of the European market suggests a clinical testing
products market in excess of 4000 million US$ in the 1990s (Biomedical
Business International). In the US, the current biosensor market is already
reported to be 12 million dollars, and future prospects vary from 100 to 10,000
million dollars by the turn of the century. This compares with a world market in
1985 of 1.5 billion dollars with an estimated growth rate of 9.5%, achieving a
world market of 2 billion in 1990 and then expanding upwards and outwards.
Need for Biosensor
Other Markets. Among the market shares, nearly 50% belongs to the medical
arena (Technical Insights Inc.) with veterinary and agricultural applications
amounting to a figure of half the size.

Table 1. Fifteen characteristics required in a commercial sensor

Relevance of output signal to measurement environment
•Accuracy and repeatability
•Sensitivity and resolution
•Dynamic range
•Speed of response
•Insensitivity to temperature (or temperature compensation)
•Insensitive to electrical and other environmental interference
•Amenable to testing and calibration
•Reliability and self-checking capability
•Physical robustness
•Service requirements
•Capital cost
•Running costs and life
•Acceptability by user
•Product safety-sample host system must not be contaminated by sensor
 Biosensor Market

USA market projection for biosensors

The biosensors market is expected to grow from $6.72 billion in 2009 to $14.42 billion
in 2016 (Yahoo Finance).
How to start a biosensor company
Biosensors are getting really popular. For the newbie, biosensors are like the machines which are
trained to recognize biological responses and convert them into mechanical signals. One of the best
examples of biosensors is blood sugar monitors which are capable of detecting blood sugar levels
almost instantly and accurately.
Biosensors are one of the most innovative and exciting fields now in the market for healthcare.
Irrespective of the weak economy and doubtful future outlook most industry experts are predicting a
60% annual growth rate for the biosensor industry and most of the drive comes from health-care and
related industries. There is also an acute shortage of donated livers, pancreas and pancreatic cells for
diabetics as well as kidneys for transplants but several companies are racing to provide artificial
substitutes which combine human cells with innovative engineering as standby or bridge transplants!
Innovative medical biosensor devices which can be used as bridges till organs become available are
now the target of several biosensor companies! Several other industries have also turned their
interests towards the use of biosensors like the food industry for food quality appraisal as well as
environmental agencies for rapid and accurate environmental monitoring. Several surveys have
estimated a surge of about £12,000,000,000 per year in the healthcare industry alone.
How to start a biosensor company
Putting up a biosensor company however will require extensive research and development into this
extremely technology sensitive field. The research required to innovate biosensors is extremely original
and involves several disciplines of physics, biochemistry, human anatomy, laboratory sciences and
bioreactor science, physical chemistry, human bodily reactions, electrochemistry, electronics and software
engineering. As a result you will need trained staff of the highest caliber to work on your research and
development wing to get the best biosensors out in the market for different purposes. The current market
is flooded with potentiometric as well as amperometric biosensors which use special colorimetric paper
enzyme strips. The potential for more and better sensors and substitute organs is a boon for patients who
are suffering life-threatening diseases.
Constant innovation will require highly paid and talented staff and you will also require a precision
modern fully automated factory to produce the technique sensitive biosensors for marketing and sales!
The ideal biosensor if you manufacture should be tiny, swift, easy to handle, cheap and easily available. It
should last for a long time and should be able to be stored in different weather conditions. The
possibilities of having your own patented innovative design can result in several more biosensors based
on the same research model and used for different detection purposes.