2407 Unit 6 Nanotech For Biophotonics 12 - 26

• As it is, they are not biocompatible
• They are covered with materials like SiO2 (silica) which is biocompatible.
• Attach this with bio molecules, e.g. proteins.
• Inject the aqueous solution into body
• The biomolecules take care of delivering the particle in the exact location in the body.
• Take the fluorescence image.
• optical properties of the quantum dots did not change after conjugations and solubilization.
• Qdot emission was 100 times as stable as that of the common organic dye rhodamine 6G
against photobleaching
Protein-attached Qdots
• They were biocompatible in vitro as well as with living cells
• For this they transferrin-Qdot bioconjugates were used.
• Cultured HeLa cells were incubated with mercapto-Qdots as control and with transferrin-QD
bioconjugates.
• Only the transferrin-Qdot bioconjugates were transported into the cell, as evidenced by
emission from the stained cells, indicating receptor-mediated endocytosis.
• Thisresult was taken as evidence that the attached transferrin molecules were still active and
were recognized by the receptors on the cell surface.
ZnS-capped CdSe Qdots

They were coated with a lung-targeting peptide accumulate in the lungs of mice after
intravenous injection, whereas two other peptides directed Qdots to blood vessels or
lymphatic vessels in tumors.
• Chimeric fusion protein was electrostatically bind to the oppositely charged surface of
capped colloidal core-shell-type CdSe–ZnS quantum dots to produce a bioconjugate.
• In this approach, the capped CdSe–ZnS nanocrystals, first with alkyl- COOH capping
reagents. Adjusting the pH (basic) of an aqueous dispersion produces the quantum dots
with negative charges (—COO- groups).
• The use of an engineered bifunctional recombinant protein consisting of a positive charge
domain leads to the formation of a bioconjugate by electrostatic attraction.
• It was proposed that the use of a porous microbead of polystyrene to capture quantum dots
in specific quantities and in a wide range of colors and intensities.
• The application to DNA analysis by preparing microbeads of three different colors and
attaching them to strips of genetic materials, each color corresponding with a specific DNA
sequence. Then they were used to probe complementary pieces of genetic material in a
DNA mixture.
METALLIC NANOPARTICLES AND NANORODS FOR BIOSENSING
Biosensor
Metal nanoparticles and nanorods are used for biosensing
Biosensors are analytical devices that can detect chemical or biological species or a
microorganism
1. Bioreceptor 2. Transducer & 3. Detector

Biosensing using Metal Nanoparticles
• Metal nanoparticles, such as silver (Ag), gold (Au), platinum (Pt) and palladium
(Pd) have attracted much interest in the construction of biosensors due to their
 Large surface-to-volume ratio.
 High surface reaction activity.
 Strong adsorption ability to immobilize the desired biomolecules and
 The unique optical (SPR) and electromagnetic properties.
SURFACE PLASMON RESONANCE (SPR)
• Surface plasmons are electromagnetic waves that propagate along the interface between a
metal and a dielectric material.(collective oscillations of free electrons known as surface
plasmons).
• When polarized light hits a metal film at the interface of media with different refractive
indices, surface plasmons are generated and this phenomenon known as Surface plasmon
Resonance(SPR).
9
Localized Surface Plasmon Resonance (LSPR)
 d << 
Nanoparticle
d
SPR in nanometer-sized structures is called localized surface plasmon resonance(LSPR)

How SPR used in Sensor
• The excitation of LSPR in metal nanoparticles by electromagnetic irradiation
results in strong light scattering and a characteristic plasmon band.
• For sensor applications, the change of the local refractive index by binding of
biomolecules to the particle surface causing a detectable shift of this plasmon band.
• The intensity and position of this band depends on ,
• size,
• shape,
• material and
• material composition.
Mechanism
A biosensor in general utilizes a biological recognition element that senses the presence of an
analyte (the species to be detected) and creates a physical or chemical response that is
converted by a transducer to a signal.
Fig:-General scheme for biosensing.
6
Surface Plasmon Resonance Detection


1
reflectance
detector
0
c o 1
angle
Au
analyte
Change of the local refractive index by binding of biomolecules to the particle surface causing
a detectable shift of this plasmon band.
Gold Nanoparticle
SPR Excitation of Au Nanoparticles
Sensing DNA Hybridization
AuNPs using functionalized thiols which can helps

to attatched.
• A single-stranded DNA modified with a thiol group at one terminal, to a gold

nanoparticle----- Red color
• But DNA attached to the gold particle hybridizes with the complementary DNA in
the test sample----- Blue black
UP-CONVERTING NANOPHORES
(Inorganic Nanoparticles)
• Another group of nanoparticles useful for bioimaging as well as for light activation
• of therapy is that of rare-earth-ion-doped oxide nanoparticles
• The rare-earth ions are well known to produce IR to visible upconversion by a number of
mechanisms as shown in Figure 15.6.
• These upconversion processes in rare-earth ions, like the two-photon absorption in organics
are quadratically dependent on the excitation intensity.
• Thus, they provide better spatial resolution.
• They produce background-free (practically no autofluorescence) detection, because the
excitation source is in the near-IR (generally 974-nm laser diodes).
Difference between Up-conversion in nanoparticles
and Two-photon absorption in organic dyes
rare-earth-ion-doped oxide nanoparticles Two-photon absorption in

organic dyes
The up-conversion process in the rare-earth the two-photon absorption in
nanoparticles is by sequential absorption organic dyes is a direct (simultaneous)
through real states and is thus considerably two-photon absorption through a
stronger virtual state
Therefore, a low-power continuous-wave Therefore, it requires a high-peak-power
diode laser at 974 nm (which is also very pulse laser source
inexpensive and readily available) can be
used to excite the up-converted emission
the emission from the rare-earth ion is a the emission from dye is fluorescence
phosphorescence with a lifetime typically in with a lifetime in nanoseconds
milliseconds
Applications that require short lived fluorescence cannot use the phosphorescence from these
up-converting nanoparticles, also referred to as nanophores or nanophosphors

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• As it is, they are not biocompatible

ZnS-capped CdSe Qdots

1. Bioreceptor 2. Transducer & 3. Detector

SURFACE PLASMON RESONANCE (SPR)

SPR in nanometer-sized structures is called localized surface plasmon resonance(LSPR)

Surface Plasmon Resonance Detection

AuNPs using functionalized thiols which can helps

• A single-stranded DNA modified with a thiol group at one terminal, to a gold

rare-earth-ion-doped oxide nanoparticles Two-photon absorption in

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