64 Chapter 2 Nanomaterials in biomedical diagnosis

synthesized with chemical reactions. The hetero fullerenes are

the fullerenes containing one or more carbon atoms, which are
replaced by other atoms such as nitrogen, boron, etc.

2.2.4.9 Metal-based nanomaterials

Metal-based nanoparticles are used in many applications
such as sunscreens and cosmetics, as antibacterial agents, and
for diagnostic and therapeutic purposes. Metal-based nanoma-
terials include silver nanomaterials, gold nanoparticles, and
metallic and nonmetallic nanoparticles [29]. In the bioanalytical
field for medical diagnosis, gold nanoparticles are used at a
higher rate because they can be easily synthesized, show higher
biocompatibility, and are nontoxic. These nanomaterials can
also be used as labeling agents as well as biosensing agents.
Gold nanomaterials can be used for light-based techniques
because they have higher scattering properties, strong absorp-
tion, and plasma resonance. The ease of surface modification of
gold nanoparticles by using a variety of biomolecules, like oligo-
nucleotides and proteins, to create specific/selective binding,
enables for the development of new biosensor platforms with
enhanced capabilities in the detection of various biological/
chemical analytes. Silver nanoparticles are helpful in the sepa-
ration of bacteria, viruses, and other blood components, facili-
tating clearer Raman spectroscopy signals of the components.
This method further helps in identifying the bacteria and
viruses in a much shorter time span [30].

2.3 Approaches to nanomaterial production

The nanomaterials have opened, and will continuously
unlock, synchronous and unpredictable domains of their appli-
cations with the development of newer technologies.
The development of nanofunctional devices depends on the
controlled assembly of nanoscale objects for particular applica-
tions. The two general approaches to the synthesis of nanoma-
terials and fabrication of nanostructures are top-down and
bottom-up approaches (Fig. 2.2).

2.3.1 Top-down approach

The top-down approach involves successive scaling down of
larger (macroscopic) complex bulk materials to smaller, nano-
sized materials using externally controlled physical processes
like etching through the mask, crushing, milling, or grinding.
 Chapter 2 Nanomaterials in biomedical diagnosis 65

Figure 2.2 Techniques of nanomaterial synthesis. *NMs: nanomaterials.

Severe deformities within the surface structure during the top-

down approach show the impact on physical features and sur-
face properties of nanomaterials [31]. Top-down methods are
costly, slow, and are not well-suited for large-scale production.

2.3.2 Bottom-up approach

The bottom-up approach assembles materials from an atom
and molecule scale to form larger structures. It is mostly used
for developing nanoscale materials of uniform size as well as
shape. It eventually involves the synthesis of nanomaterials via
self-assembly and molecular patterning chemically, along with
an effective means of restricting the particle growth.
Nanomaterial fabrication is much less expensive using the
bottom-up approach [31].
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2.4 Nanomaterial method of synthesis

Nanomaterials can be synthesized using various physical,
biological, or chemical methods. Commonly used methods of
nanomaterial synthesis include.

2.4.1 Physical method

2.4.1.1 Inert gas condensation
This technique involves fabrication of nanoparticles via
evaporation of a metallic source (titanium, cadmium, silver,
magnesium, etc.) with the help of an inert gas (argon or helium)
[32]. The inert gas condensation process is widely used in the
synthesis of ultrafine metal particles. It involves the evaporation
of a metallic source by induction heating, followed by the cool-
ing of metallic vapors with inert gas. The metal core transfers
heat energy to cooler inert gases via diffusion and convective
heat flow. The evaporated atoms smash with gas atoms within
the evacuated compartment and lose their kinetic energy. This
passage of inert gas facilitates nucleation, a collision bonding
process, and ultimately leads to the collection of particles by a
deposition method onto a cold surface [33]. The size of the par-
ticles depends on the inert gas pressure, temperature, ablation
rate, sputtering technique, and gas composition [34].

2.4.1.2 Plasma arc discharge

The plasma arc discharge method is utilized for the synthesis
of low-dimensional carbon materials (e.g., graphite, nanodia-
monds) between graphite electrodes in an inert environment.
The process involves vaporization of substrate carbon content
in the presence of a catalyst (e.g., iron, nickel, cobalt, yttrium,
boron, gadolinium, etc.) under a reduced atmosphere of inert
gas (argon or helium). The migration of arc (ions) between two
graphite electrodes leads to the formation of plasma composed
of a mixture of carbon vapors, inert gas (helium or argon), and
catalysts. The erosion of anodes due to the transfer of energy of
vaporization of arc to anode imparts high CNT production [35].
The nucleation and deposition of carbon clusters requires
suitable temperatures and space domains. However, the nucle-
ation, as well as the growth process, is influenced by the size
and spatial distribution of catalysts in the anode. The outcome
of the plasma arc discharge process is also influenced by several
variables like buffer gas, current, voltage, catalysts, electrode
morphologies, carbon sources, electrical field, etc. [36,37]. This
 Chapter 2 Nanomaterials in biomedical diagnosis 67

method is considered suitable for production of carbon nano-

particles and graphene at large scale. It is a cost-effective pro-
cess and doesn’t require expensive equipment.

2.4.1.3 Thermal plasma jets

Thermal plasma jets are also called thermal plasma torches,
and are created by electric arc heaters [38]. Direct current and
radio frequency plasma torches are widely used for nanomater-
ial production. The direct current plasma (DC plasma) torch
consists of cathode and anode nozzles of either rod or hollow
typed electrodes. A rod-shaped cathode is made up of thoriated
tungsten and is the most sensitive to oxygen-containing gases
and prone to severe erosion. However, hollow-shaped electrodes
are made up of oxygen-free, high-purity copper. In this method
an electric arc is first introduced at high voltage between elec-
trodes and hot gases are flown around the column to maintain
a temperature of 8000 16,000K through the heat exchange,
conductive, and convective processes. A nozzle is placed at the
end of the torch to expand the heated gas and form a plasma
jet. The jet conditions (temperature, velocity etc.) are governed
by nozzle design and can be customized for the fabrication of
nanomaterials.
A radio-frequency plasma (RF plasma) torch is composed of
a confinment tube and an electromagnetic coil. The conserva-
tion of plasma in an RF plasma torch can be achieved via an
induction heating mechanism where the alternating magnetic
field is generated by oscillating current inside the confinement
tube containing an electrically conducting medium. Eddy cur-
rents generated under alternating magnetic field in the medium
dissipates heat via the Joule heating process. The energy so
evoked is enough for the continuous ionization of injected gas.
The RF plasma torch usually provides a larger volume of heat-
ing zone, which ultimately provides a higher residence time of
feedstock in the plasma zone, leading to a significant enhance-
ment of material growth. However, relatively low power effi-
ciency (, 70%) is still a big challenge to be addressed before
this technique can be used on large scale for fabrication of
nanomaterials [39,40].

2.4.1.4 Ion sputtering

The ion sputtering technique involves the bombardment of
target material with ions (sputter atoms) at a suitable electron
volt. It is based on two different mechanisms: thermal vaporiza-
tion theory (vaporization of target material due to energy
68 Chapter 2 Nanomaterials in biomedical diagnosis

transfer by bombarding ions), and momentum transfer theory

(emission of surface atom of target when kinetic moments of
incident particle are transferred to target surface atom) [41].
Sputtering can be performed directly, under vacuum, using
radio frequency plasma jets, or in a magnetron. Ion sputtering
has found wider applications in the fabrication of thin film,
multiple layers of metal and metal oxides, surface cleansing,
and surface layer analysis [42].

2.4.1.5 Laser ablation

In the laser ablation method, the intense pulse of laser beams
of temporal width interact with material at a constant time. The
laser energy is then transmitted to the lattice via electron-proton
coupling. The energy of the laser beam continues to pass into
the material even after the carrier has finished transferring its
energy and when pulse width is more than proton coupling time.
This results in dissipation of laser energy as heat from the mate-
rial’s surface where the laser beam is incident and causes defor-
mity in material like formation of a pronounced heat affected
zone, recast layers, surface debris, mechanical cracks and other
defects. On the other hand, if the pulse width is smaller, the laser
energy is constrained to its initial volume, in which irradiation
was absorbed. In this case, there is no dissipation of laser energy
into heat from the irradiated area, and due to the ultrashort
pulse width, the surface enthalpy of the material is of higher
magnitude than its sublimation enthalpy, which facilitates abla-
tion of material [43]. Single walled carbon nanotubes (SCNTs)
with excellent structural integrity and high purity can be pro-
duced by pulsed laser vaporization method [44].

2.4.1.6 Laser pyrolysis

Laser pyrolysis is a viable platform to develop nanoscale
materials for wider applications. The process involves collision
of a continuous wave of 10.6 μm wavelength CO2 laser beam
with a stream of reactants fed through a nozzle into the reactor.
The process of laser pyrolysis proceeds as emission of the con-
tinuous wave of CO2 laser and an infrared absorption band of
components of the reactant stream resonates with the subse-
quent transfer of energy to nonlaser active molecules [45].
Precursors suitable for laser pyrolysis exhibit the following
properties:
1. Directly produce desired products upon decomposition [46].
2. Materials that decompose upon absorption of energy with-
out producing product like SF6.
 Chapter 2 Nanomaterials in biomedical diagnosis 69

3. Species facilitating absorption, decomposition, and reacting

with reactant molecules like production of vanadium oxide
from vanadium oxy-trichloride vapor by ethylene (organic
ligand) in existence of oxygen [47].

2.4.1.7 Ball milling

Ball milling is one of the fabrication methods used for metallic
and ceramic nanomaterials. Ball mills use hardened steel or tung-
sten carbide balls as a milling tool. A container is rotated on its
own axis, as well as on a central axis. In order to ground material
into a fine powder, the speed of rotation along with the duration
of milling needs to be regulated. The use of heavy milling balls
exhibits high impact of attrition and energy of collision with a
subsequent rise of temperature ranging from 100 C to 1100 C in
the container. The efficiency of milling is dependent upon the full-
ness of the container. Usually a less than half-filled container with
2:1 mass ratio of balls to material is advisable for an efficient ball
milling process. The major drawback is contamination of product
with ball material, which can be hindered by filling the remaining
space in the container with inert gas or air [48].

2.4.1.8 Chemical vapor deposition

The chemical vapor deposition (CVD) process is relatively
cost effective in comparison to the other processes. It involves
the exposure of a substrate maintained at a high temperature to
a volatile precursor. The reactant breaks into different products,
which diffuse on the surface, undergo chemical reaction,
nucleate, and grow to form the material of choice. It is a hybrid
technique for coating organic, as well as inorganic, materials
[49]. A variety of CVD techniques are available, including metal-
lo organic CVD [50], molecular beam epitaxy [51], vapor phase
epitaxy, and plasma enhanced CVD.

2.4.2 Biological/green methods

Nanoparticles are diversely used as a catalyst, an adsorbent,
as sensors, and as ferrofluids for synthesis of nanomaterials,
which have wider applications to develop devices for diagnostic
purposes. Shape and size of the materials can be well con-
trolled. However, the shape is variable depending on the
amount of polymeric stabilizer, reaction condition, and oil/
water/surfactant ratio. Nanoparticles of various metals are syn-
thesized using plant extracts [52], bacteria [53], fungi [53], and
yeast [54] as precursors.
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2.4.3 Chemical methods

2.4.3.1 Hydrothermal synthesis
Material synthesis via hydrothermal methods involves the
direct crystallization from solution in two steps: nucleation fol-
lowed by crystal growth. The size and morphology of the mate-
rials crystallized are governed by supersaturation, which is
affected by various processing variables like temperature, pH,
reactant concentration, etc. Hydrothermal methods are exe-
cuted in a pressure vessel or autoclave with a Teflon or alloy lin-
ing. The temperature of the autoclave can be increased
progressively above the boiling point by increasing the vapor
pressure of the compartment. Hydrothermal synthesis can con-
trol particle size, morphology, crystalline phase, and surface
chemistry by regulating temperature, pressure, additives, time,
and solvent properties [55]. TiO2 nanoparticles can be prepared
by hydrothermal reaction in two steps: the first step involves
formation of titanium hydroxides by hydrolysis, and the second
step is the formation of titania by dehydration or a condensa-
tion reaction using water as a solvent [56].

2.4.3.2 Solvothermal synthesis

Solvothermal synthesis to achieve crystal formation is similar
to hydrothermal synthesis except that a variety of organic sol-
vents other than water are used. It involves heating an organic
solution and surfactant above the boiling point of the solvent
(,250 C) to facilitate the formation of supersaturated solutions
followed by nucleation of crystals with crystal growth at a faster
rate. Surfactant use prevents lump formation during crystal for-
mation. However the catalyst plays an important role in initiat-
ing the growth of crystals of inorganic material. This method
can control shape, size distribution, and crystallinity better than
the hydrothermal method [55]. TiO2 nanoparticles [57], nanor-
ods, and NWs are usually prepared by solvothermal synthesis.

2.4.3.3 Cryochemical synthesis

The cryochemical synthesis method involves crystallization
of various solutions at cryogenic temperatures followed by the
removal of the solvent using freezing, drying, or spraying
(Fig. 2.3). Cryochemical methods are technologically and eco-
logically safe and offer high output at low cost [58]. It is per-
formed in various steps.
 Chapter 2 Nanomaterials in biomedical diagnosis 71

Preparation of dispersive system

Freezing

Solvent removal
by sublimation

Crystals obtained

Thermal treatment Desublimation

De-aggregation of granules

Figure 2.3 Flowchart of cryochemical synthesis.

2.4.3.4 Aerosol-based process

An aerosol-based process is a dynamic field of particle tech-
nology used to produce nanosized materials. The assembly
used to produce aerosol consists of three main parts: an atomi-
zation chamber, a tubular reactor, and a particle collector. In
the atomization chamber high-energy impact carrier gas com-
bined with a liquid solution generates fine aerosol. Aerosol gen-
eration is usually achieved by pumping a precursor liquid via a
peristaltic pump at a flow rate of 2 3 mL min21. The generated
aerosol is transported into the reaction chamber and main-
tained at pyrolysis temperature of 800 C 1000 C. The particles
undergo thermolysis at a high temperature to form microporous
particles, which is followed by conversion into dense particles
by the sintering process. Through the intermediate tube con-
necting reactor chamber to particle collector unit, dry air is usu-
ally introduced at a high pressure and high flow rate as
quenching gas near the outlet of the reaction chamber. This
process provides better control of the particle size [59].

2.5 Characterization of nanomaterials

Properties of nanomaterials vary according to their size, and
thus their physical, structural, and optical properties need to be
characterized. Various techniques used for characterization
(Table 2.1) are discussed in the following.