Review Three Synthesis Methods of CDX

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Published in IET Nanobiotechnology
Received on 21st August 2012
Revised on 26th November 2012
Accepted on 29th November 2012
doi: 10.1049/iet-nbt.2012.0028
ISSN 1751-8741
Review: three synthesis methods of CdX (X = Se,

S or Te) quantum dots
Samad Mussa Farkhani, Alireza Valizadeh
Department of Medical Nanotechnology, Faculty of Advanced Medical Science, Tabriz University of Medical Sciences,
Tabriz 51664, Iran
E-mail: a.valizadeh_medicalnano@yahoo.com
Abstract: Quantum dots (QDs) are one of the rst nanotechnologies to be integrated with the biological sciences that used for
imaging or tracking macromolecules/cells in cell/tissue. Because of QDs are important in biomedical and biological applications,
identify a variety of synthesis methods to produce QDs with different characteristics also is particularly important. Hence, in this
review the authors discussed three methods for synthesis of heavy metal chalcogenide-based QDs for use in biomedical eld: (i)
Organometallic method for synthesis of QDs consists of three components: precursors, organic surfactants and solvents. The
authors also discussed water-solubilisation strategies of synthesised QDs including encapsulation and ligand exchange. (ii)
Aqueous synthesis technique using short-chain thiols as stabilising agents is a useful alternative to organometallic synthesis of
CdSe, CdS and CdTe QDs. (iii) The third method discussed in this article for QDs synthesis involves the utilise of
microorganisms to prepare QDs with controlled size, shape, chemical composition and functionality. The authors also
discussed recently new methods for the synthesis of the appropriate QDs for use in biology. In addition, attachment of
biomolecules such as antibodies, oligonucleotides on the surface of QDs for specic targeting and different opinions about
toxicity of QD have been studied.
Nomenclature
QDs
TOPO
TOP
HPA
TDPA
ODE
TBP
OA
(TMS)2 S
ZnEt2
DTA
(BDMS)2 Te
HAD
TEM
HRTEM
Cd (Ac)2
DDA
BSA
TGA
GSH
MSA
APTES
AOT
CA
quantum dots
trioctylphosphine oxide
trioctylphosphine
hexylphosphonic acid
tetradecylphosphonic acid
octadecene
tributylphosphine
oleylamine
hexamethyldisilathiane
diethylzinc
dowtherm A
bis(tert-butyldimethylsilyl) telluride
hexadecylamine
transmission electron microscopy
high-resolution transmission electron
microscopy
cadmium acetate
dodecylamine
bovine serum albumin
thioglycolic acid
glutathione
mercaptosuccinic acid
(3-aminopropyl)triethoxysilane
dioctyl sulphosuccinate sodium salt
(aerosol-OT)
cysteamine
IET Nanobiotechnol., 2014, Vol. 8, Iss. 2, pp. 5976

doi: 10.1049/iet-nbt.2012.0028
TAA
TP
PEG
MAA
PLQY
thioacetamide
tiopronin
polyethylene glycol
mercaptoacetic acid
photoluminescence quantum yield
Introduction
Interest in colloidal inorganic nanoparticles such as

semiconductor
nanocrystals,
metal
and
magnetic
nanoparticles has stably grown in the past two decades
because of their unique optoelectronics, magnetic and
electrical properties that are different from their bulk
structure counterpart [113]. Among the various colloidal
semiconductor
nanocrystals,
representatively
metal
chalcogenide nanocrystals, have been interested because of
their quantum connement effects and size-dependent
photoemission characteristics [14]. For biological imaging,
Cd-based chalcogenide (This term generally refers to
sulphur (S), selenium (Se), tellurium (Te) elements of the
periodic table) nanocrystals are regarded to be promising
materials [11]. Quantum dots (QDs) are semiconductor
nanoparticles including of elements from the periodic
groups IIVI or IIIV and their size are ranging between 2
and 10 nm in diameter. This ranging of diameter is close to
or smaller than the dimensions of the exciton Bohr radius.
Moreover, the mobility of charge carriers (electrons and
holes) is restricted within the nanoscale dimensions, and
59
& The Institution of Engineering and Technology 2014
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this quantum connement effect QDs with unique optical and
electronic features [15]. Owing to this size-dependent
property, the absorption and emission properties [16] of the
semiconductor QDs can be easily tuned by controlling the
particle size, shape or surface structure [17]. Fig. 1 shows a
schematic drawing representing the wavelength of light
emitted by QDs is tunable by changing the particle size.
Compared with traditional organic uorophores, QDs
process several advantages in uorescence properties for
biological applications, that including: a high brightness
because of the extinction coefcient and quantum yield,
broad absorption characteristics and a narrow band width in
emission spectra, independence of emission on the

excitation wavelength, continuous and tunable emission
maxima because of quantum size effects, and a longer
uorescence lifetime ranging from 10 to 40 ns, and stability
under relatively harsh environments [10, 18]. QDs with
careful functionalisation have been widely used for imaging
and sensing [6, 10, 1927] and tracking particles or cells
[2830] in biology and medicine. Because of these features
mentioned and the importance of medical and biological
applications of QDs, in this review, we have discussed the
three synthesis methods and also new method of CdX (X =
Se, S or Te) QDs, which is the most common commercially
Fig. 1 Schematic drawing demonstrated

a Exciton has shown when a photon is absorbed by a semiconductor
b Wavelength of light emitted by QDs is tunable by changing the particle size and their emission wavelength is narrow and symmetrical
c Series of QDs with different core sizes and emission wavelength can be excited simultaneously by a single excitation light source
60

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available and used as secondary antibody conjugates for
labelling [31]. In addition, we briey discussed the
applications, toxicity, photostability of QDs and surface
engineering of QDs for use in biology.
QD materials
QDs are constructed from semiconducting materials. The

particles are generally made from hundreds to thousands of
atoms of group II and VI elements (e.g. CdSe and CdTe) or
group III and V elements (e.g. InP and InAs) [15]. QDs
composed of Group IIVI have recently drawn signicantly
interest because of their potential applications in
optoelectronics and biomedicine elds such as photovoltaic
cells and biomedical tags [3238]. Among the various QDs,
colloidal IIVI group metal chalcogenide QDs such as CdS,
CdSe and CdTe, have been studied intensively on their
quantum connement effects including size-dependent
photoemission properties, because of the latest development
in synthesis leading to high-quality QD ensembles with
narrow size distribution [39]. They are most widely utilised
QDs for biomedical eld to date [40]. Table 1 listed key
parameters of bulk semiconductors usually used for CdSe,
CdTe and CdS QDs.
Design principles of QDs for biological uses
QDs modelling for biological use includes: (i) Synthesise the

QDs: most QDs for biological environmental are based on
heavy metal chalcogenide compounds, such as CdSe and
CdTe, which can leach heavy metals in tissue. (ii) To solve
the problem of mentioned in number one, a non-heavy metal
shell, such as ZnS or CdS is used as a barrier. (iii) Adjusting
its surface so that it is biologically compatible, and then
further modifying the surface so that the QD can be directed
to a target. There are three primary ways to target a
biocompatible QD: with antibodies, peptides or small
molecules [42]. In this review, we discussed the conventional
and new methods to the synthesis of water soluble QDs.
QD structure
QDs for use in eld of biology consist of:

1. Core of QD: The central part of QDs that determines the
optical properties of the ultimate structure. The typical QDs
consist of a IIIV, IVVI, or IIIV semiconductor core (e.g.
CdTe, CdSe, PbSe, GaAs, GaN,InP and InAs) [6].
2. Shell of QD: Core of QDs has an inorganic shell. Coating
around the core is a wide band gap inorganic semiconductor
shell such as CdS or ZnS in order to minimise the surface
deciency and increase the quantum yield (QY) [43].
3. Water-soluble QD: These QDs are hydrophilic and are
soluble in water and usable in biological buffers. QDs for
use in biology need to be water-soluble. There are two
methods for making QD water-soluble: (1) Ligand
exchange (2) Encapsulation. Normally, QDs synthesised in
organic solvents have hydrophobic surface ligands such as

trioctylphosphine (TOP), trioctylphosphine oxide (TOPO),
tetradecylphosphonic acid (TDPA), oleic acid or et. In
method one these ligands of QDs surface could be replaced
by some water-soluble ligand such as thiol-based molecules
(such as mercaptocarbonic acids), peptides and other. This
is ligands exchange. QDs can also be encapsulated by a
shell of material such as silica, phospholipids, amphiphilic
polymers (poly(acrylic acid), polyethylene glycol (PEG)) to
become biocompatible and makes them more soluble in
aqueous media [18].
4. QD bioconjugate: A water-soluble QD bound to an
afnity molecule such as antibodies, peptides [44].
QD applications
The application of QDs, as a new technology for biosystems,

has been mostly studied on mammalian cells. There is an
increasing tendency to apply QDs as markers in medicine
and biology that because of their small size, brightness,
independence of emission on the excitation wavelength and
stability under relatively harsh environments [45]. They also
have excellent photostability [18] and overcome the
limitations associated with photobleaching and because of
the small structures of QDs, some physical properties such
as optical and electron transport characteristics are quite
different from those of the bulk materials [46]. Some of the
application summarised in Fig. 2 [45].
6 Stability of QDs in live cells and

photostability
The right choice of QDs diameter and monolayer structure, is
important for bioapplications of QDs. Zhu et al. [47]
discovered that by increasing the diameter of the QDs,
monolayer stability decreases. They showed that the
smallest QD studied, (2.9 nm core diameter), demonstrates
high stability in cells, against the largest QD (5.9 nm core
diameter) loses 70% of its monolayer over the 24 h. They
also showed that, when a dithiolate monolayer is replaced
by a monothiolate monolayer, the monolayer stability
decreasing by 40%. They found that QD monolayer
stability is correlated with both QD diameter and monolayer
structure, with right choice of both particle size and ligand
structure required for intracellular stability. Since the
synthesis process of QDs effect on particle size and
monolayer structure, it is important that we fully understand
the procedures and process of QDs synthesis. Also the
photostability is one of most important properties of QDs
for cell imaging, particularly for the long-term monitoring.
Ma et al. [48] studied the photobleaching of naked CdTe
QDs and bovine serum albumin (BSA) coated CdSe/CdS/
ZnS QDs under single-photon excitation and two-photon
excitation. Based on their research the BSA-coated core/
shell QDs had improved the photostability up to 45 times
than the naked QDs because of the shielding effect of the
QD shell. Manner et al. [49] discovered that the properties
Table 1 Important parameters of bulk semiconductors usually used for CdSe, CdTe and CdS QDs [41]
Material
CdSe
CdTe
CdS
Structure, 300 K
Type
Egap, eV
Lattice parameter, [A]
Density, kg/m3
wurtzite
zinc blende
wurtzite
IIVI
IIVI
IIVI
1.74
1.43
2.49
4.3/7.01
6.482
4.136/6.714
5810
5870
4820

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Fig. 2 Demonstrated some of QDs applications in medicine and biology [45]
of environment have signicant impact on the stability of

QDs, and the improvement of their resistance to
photooxidation may require the control of the properties of
the surrounding solvent or matrix.
Synthesis QDs
In the early 1980s, several pioneering groups began to work on

QDs [50]. Ekimov et al. in 1982 discovered the size-dependent
characteristics of ionic nanocrystals, in a glass matrix, at the
loffe Institute in St. Petersburg, Russia [51, 52]. L. Brus in
1983, reported size-dependent behaviour in colloidal CdS
nanocrystals through the measurement of size-dependent
redox potentials [53]. QDs Synthesis was rstly
accomplished by precipitation from a solution containing the
metal ion (Ag, Hg, Pb, Zn, Cd, In) by a hydroxide of S, Se
or Te followed [54]. The preparation process includes control
of the shape, size, homogeneity and the surface structure of
the QDs and this is a very important part in producing those
mentioned unique properties [55]. In this review we
discussed three popular methods and also new methods for
synthesis of QDs used for biology:
1. Organometallic synthesis: this method is the most popular
method for QDs synthesis.
2. Aqueous route for synthesis QDs: low-temperature reaction
3. Biosynthesis of QDs
4. New methods for synthesis of water-soluble QDs
7.1
Organometallic precursor synthesis of QDs
With entrance of nucleation and growth techniques to

synthesize QDs in high-temperature organic solvents,
62
synthesis of QDs made easier and more controllable. An

organometallic system for synthesis QDs consists of three
components: precursors, organic surfactants and solvents.
Sometimes, surfactants also serve as solvents [56]. Metal
chalcogenide-based QDs with different sizes to be
manufactured by alteration of the amount of precursors and
crystal growth time; this is important for their uorescent
properties in that their emission wavelength is dependent
upon crystal size, and hence tuneable during manufacture
[57]. In this process to create QDs, ionic sources of the
component materials such as Cd2+ are needed. This method
consists of pyrolysis of organometallic precursors into a hot
coordinating solvent to manufacture monodisperse (<5%
size dispersion) QDs composed of cadmium chalcogenides
[1]. Organometallic synthesis of QDs, invented in the early
1990s by Steigerwald and Brus [58], developed to a
practical level and for rst time controllable nucleation and
growth of narrow size distribution QDs was demonstrated
by Murray et al. in 1993 [59]. Precursors describe the
combination of the particles that are formed. When
precursors dissolved in the solvent, they decompose into
reactive species, known as monomers [60]. Steigerward
indicated that an important step to synthesis of QDs is
identifying suitable precursor molecules such as
organometallic compounds: a suitable precursor for the
synthesis QDs at the required growth temperature need to
quickly decompose to yield reactive atomic or molecular
species (the monomer) that can after that contribute to
nanocrystal nucleation and growth. The most famous
example is the use of dimethyl cadmium (Cd(CH3)2) and
trialkyl phosphine selenide to yield CdSe [61, 62]. The best
been used precursors are simple molecules with leaving
groups that readily depart to leave behind the desired
reactive specie [56]. Murray et al. use Cd(CH3)2 as the
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cadmium precursor for the synthesis highly crystalline CdSe
QDs. Injection of Cd(CH3)2 into a hot solution can yield
supersaturation, nucleation, and subsequent growth but Cd
(CH3)2 is very toxic, unstable at room temperature and air
sensitive [63]. To solve this problems Peng and co-workers
in 2001 reported the use of CdO complexed to
hexylphosphonic acid (HPA) or TDPA, as an alternative to
Cd (CH3)2, in TOPO syntheses of QDs [64].
In synthesis of QDs, tow common events should occur: (1)
The nucleation: precursors at a high-temperature decompose
or react to form a supersaturation of monomers followed by
a burst of nucleation of nanocrystals (2) these nuclei growth
from molecular precursors (Fig. 3a). This method requires
accurate temperature control to separate the nucleation and
growth of the QDs, which solely depend on the temperature
of the mixture, injection process utilized as well as the
concentration gradient under control [65].
In this method, an injection temperature and a growth
temperature are selected. This high reaction temperature
(>150350C) has two major advantages: facilitates the
removal of crystalline defects and allows improvement in
the photoluminescence [66]. When the reaction medium is
warm enough, the precursors chemically convert into active
atomic or molecular species (monomers); these then form
nanocrystals and with utilise the surfactant molecules
subsequent growth is strongly affected [15]. In synthesis of
Fig. 3 Schematic diagram showing

a Cartoon depicting the stages of nucleation and growth for the preparation of
QDs based on La Mer model [1]
b Schematic drawing representing a synthetic apparatus employed in the
synthesis of QDs
doi: 10.1049/iet-nbt.2012.0028
chalcogenide QDs, cadmium precursors are organometallics

[67] such as Cd(CH3)2 while Se, S, Te precursors are often
from elemental source. Organic solvents, such as TOPO
[59], HDA [68], octadecene (ODE) [69] of which are high
boiling point solvents containing long alkyl chains and have
the ability to create high reaction temperature (200400).
Table 2. Lists common precursors, solvents and stabilisers
used in the synthesis of chalcogenide-based QDs. ODE has
several advantages compared with other solvents such as:
good dissolving power, environmentally friendlier, stability
in air, less toxicity, low cost, low melting point (below 20
C) and this makes it easier to be handled at room
temperature. Furthermore, it has a high boiling point
(approximately 360C) coupled with its inert nature to the
Se precursor, and which makes it an ideal solvent for the
growth of high-quality QDs [70]. These molecules serve
three major purposes:
1. The reaction medium for solubilise and diffuse the
nanocrystals and the reactants involved in the growth.
2. Coordinate with unsaturated metal atoms on the QDs
surface to prevent the formation of bulk QDs.
3. They can control the speed of the reaction.
To do so, the solvent molecules require attaching and
unbinding dynamically on the surface of the growing
nanocrystals. With separating one molecule from the
surface of the nanocrystal, monomers can be incorporated
into the nanocrystal, and thus it can grow [71]. Used
surfactant for synthesis QDs needs to be able to exchange
on and off the growing crystals, such that regions of the
nanocrystal surface are transiently available for growth, yet
total crystals are on average monolayer protected to block
aggregation. These molecules display tow domains: one
non-polar, usually a long alkyl chain, and a polar head
group. Non-polar tail affects the diffusion properties, while
the polar head group generally affects the binding efciency
[71]. The surface of energy with which surfactant molecules
present in the growth medium stick to the surfaces of
growing nanocrystals is one of the most important
parameters inuencing crystal growth [56]. Examples of
surfactants consist of fatty acids, amines, some
nitrogen-containing aromatics and molecules with very long
chains of carbon atoms (alkyl chains) such as alkyl
phosphine oxides, alkyl phosphonic acids and alkyl
phosphines that dynamically solvate nanocrystals [56].
Function of capping agents: with growth of nuclei, van der
Waals forces can cause quick coalescence of nuclei and
uncontrolled particle growth. Ligands such as stearic acid,
myristic acid, oleic acid and TOP are chemically bonded to
both the precursors and to the particles that form. Such
bonds resist van der Waals interactions. These molecules
may be added during the synthesis of QDs to adsorb and
restrict particle particle agglomeration, although such
molecules may, in principle, prevent monomer
sedimentation too. Additionally, it is possible that
complexation of capping agents to the active species and
precursors may hinder nucleation, which will subsequently
lead to larger, rather than smaller, particle sizes [69].
Optimisation of QDs synthesis conditions were carried out
by systematically changing the reaction condition including,
the effect of type of solvent [80, 81], form of surfactant
[78], precursor form [67], reaction temperature [82] and
working to increase the QY and monodispersity. The
reaction is normally performed in three-neck asks connect
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Table 2 Precursors, solvents and stabilisers used in the synthesis of chalcogenide-based QD
QDs
Size of
particles
Solvent
CdSe/ZnS
2.35.5 nm
TOPO
CdSeCdS
CdTe
CdSe
CdTe
CdSeCdS
CdTe
CdSe/CdS
CdS
CdSe
CdSeCdTe
CdS
CdSe/CdS/ZnS
1.211.5 nm
Precursors and stabilisers
Refrences
TOPO
CdMe2, diethylzinc (ZnEt2), TOP-Se, hexamethyldisilathiane

(TMS)2 S
(TMS)2S or (TMS)2Se or (BDMS)2Te CdMe2/TOP
[59, 73]
1.525 nm
2.57 nm
28 nm
TOPO
DDA
TOPO
Cd(Ac)2 or CdO or CdCO3, TOP-Se, stearic acid or lauric acid

CdMe2,TOP-Te
CdO,TOP-Se or TOP-Te or TOP-S ,TDPA
[73]
[74]
[75]
2.94 nm
3.8 nm
4.05 0.16 nm
ND
5.1 nm
7 nm
TOPO-ODE-DTA-T66
ODE
HDA, TOPO
TOPO
OA
HDATOPO
TOP-Se, CdO, Cd/oleic acid

myristic acid, Cd(Ac)2, S
TOP-Se, CdMe2/TOP
TOP-Se, CdMe2/TOP, H2Te, Cd(ClO4)2
CdCl2, sulphur/OA, TOPO
Cd(Ac)2, TOPSe, H2S, Zn(C2H5)2 (TMS)2 S, TOP
[76]
[77]
[78]
[79]
[14]
[68]
[72]
ND - not determined
to a Schlenk line (vacuum gas manifold) with one of the

necks. The remaining tow necks are sealed with rubber
septa and serve for the injection of reactants and the
measurement of the temperature reaction medium (Fig. 3b).
The reaction typically is performed under an inert
atmosphere, because molecules of reactant may be
pyrophoric and furthermore some types of nanocrystals may
be sensitive to air. To begin the QDs synthesis, this ask
loaded with the organic solvent and surfactant molecules,
then organic molecules are molten from powder state to a
liquid and the ask is evacuated and kept under vacuum at
150300C for 1020 min to remove volatile impurities
[71]. Elemental chalcogens (S, Se and Te) are introduced
into the reaction medium as a complex with capping agent
such as TOP or tributylphosphine (TBP). In this procedure,
the complex is produced by dissolving the S, Se or Te in
liquid TOP or TBP [71].
Organometallic precursor synthesis produces CdSe, CdS or
CdTe QDs, but has a comparatively low-QY (brightness),
usually <10%, requiring a shell of a high band-gap
semiconductor, such as CdS or ZnS to be epitaxially grown
around the core, which increases the QY up to 80%. This
layer moreover protects the core from oxidation and
prevents leaching of the Cd/Se [83].
7.1.1 Water-solubilisation strategies: QDs synthesised
with organometallic method in non-polar solutions using
aliphatic coordinating ligands (such TOPO) are only soluble

in non-polar organic solvents [84]. These QDs must
therefore be modied to become soluble in water and
compatible with the biological environment. Two general
strategies have been developed for this purpose: ligand
exchange and Encapsulation within block copolymer shells
and phospholipid micelles (Fig. 4). Ligand exchange
consists in replacing the hydrophobic ligands with new
ligands presenting anchoring groups to bind to the QDs
surface and hydrophilic solubilising groups [85]. Examples
of water-soluble bifunctional molecules used for ligand
exchange method are Aminated polymers, DHLAPEG
derivatives, Thiol--D-lactose, peptides and so on. [18, 86].
QDs generated with this method are useful for biological
environment, but ligand exchange is usually in association
with decreased QY and a tendency to aggregate and
precipitate in biological buffers. Lately, it has been shown
that these problems can be alleviated by keeping the
hydrophobic ligands on the surface, and covering the QDs
with amphiphilic molecules [84]. Examples of amphiphilic
molecules used for QDs encapsulation are phospholipid
micelles
or
amphiphilic
copolymers
such
as
-cyclodextrine, polymer microsphere and etc. [85, 86].
Coating with polymers may increase the size of the QDs by
as much as 510 nm depending on the coating. In addition,
the silica-coating method has also been used to
water-solubilisation of QDs. Hydrophilic ligands used in
Fig. 4 Schematic representation of the surface functionalisation strategy based on ligand exchange and encapsulation of the QDs within
phospholipid micelles
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Table 3 Comparison of some characteristics such as size, electronic transition energies, crystal structure and photoluminescence
behaviour of CdSe QDs capped with different stabilisers [93]
Stabiliser
crystal structure
photoluminescence
1s1s electronic transition, nm
diameter, nm
2-mercaptoethanol
1-thioglycerol
TGA
cubic CdSe (zinc blende)

trapped
360450
1.42.2

trapped
360450
1.42.2

trapped
450530
2.13.2
these two strategies may be composed of charged moieties

(carboxylic acids, amines,), however these present limited
stability and high non-specic adsorption of biomolecules
on their surface. PEG chains are more common for in vivo
applications because it decreases signicantly this
non-specic adsorption, and increases the stability and
circulating time of QDs [85, 87]. It must be considered that
many biological and physical properties of the QDs may be
affected by the surface coating, and the overall physical
dimensions of the QDs are dependent on the coating
thickness [84].
7.2
Aqueous route for synthesis QDs
Typically, CdSe nanocrystal synthesis includes the use of Cd

(CH3)2 with TOPO. The raw materials such as
organometallics, in QDs synthesis are particularly
pyrophoric, unstable, costly, toxic, and the reactions often
lack control and reproducibility [8890]. Moreover, their
surface (capped by TOPO), is hydrophobic and QDs cannot
be directly applied in bio-labelling. After the substitution of
the surface-capping molecules on QDs by hydrophilic
molecules (such as thiol groups or amines), their
photoluminescence signicantly decreased [32, 91]. Direct
aqueous synthesis of QDs such as CdS [92], CdSe [93],
CdTe [94, 95], by applying different thiol groups as
stabilising agents provides a effective alternative to widely
used synthetic routes in highly boiling organic solvents.
The use of thiols makes possible to control the kinetics of
the QDs synthesis, passivate surface dangling bonds, and
provides stability, solubility and surface functionality of the
nanoparticles [96]. In this method the ligands used during
formation of QDs become the eventual biocompatible
surface ligands and do not need an exchange process [97].
For synthesis of QDs in water directly, Vossmeyer et al.
[92] used Cd(C1O4)6H2O, H2S and 1-thioglycerol as the
precursors for the aqueous synthesis of CdS. A group of
researchers have synthesized the CdS-mercaptoacetic QD
using mercaptoacetic acid (MAA) as the stabilising agent in
the aqueous solution [98]. Using this method, CdSe

nanoparticles have been synthesised through the addition of
freshly prepared oxygen-free NaHSe solutions to
N2-saturated Cd (ClO4)2.6H2O solutions at pH 11.2 in the
presence of different thiols as stabilising agents. In this
procedure, particle size of CdSe was controlled by choosing
different types of stabilizer and through size-selective
precipitation. Table 3 summarised several characteristics of
the series of thiol-stabilised CdSe QDs with different sizes
[93].
In aqueous route, the common synthetic procedure follows
a similar route for all nanocrystals: First a metal salt (such as
Cd(ClO4)2) is dissolved in water in the presence of the thiol as
the stabilising agent. The original solution need to purge by
inert gas (mainly N gas) before the chalcogen source is
injected. The common thiols used in this procedure for
synthesis
were
2-mercaptoethanol,
1-thioglycerol,
thioglycolic acid (TGA), thiolactic acid [44, 92, 93],
1-mercapto-2-propanol and 1, 2-dimercapto-3-propanol.
Among the mentioned stabilising agents, the thioalcohols
(2-mercaptoethanol
and1-thioglycerol)
are
effective
size-regulating and stabilising agents for cadmium
chalcogenide nanoparticles [93]. Table 4 lists some of the
stabilising agents and precursor used for synthesis QDs and
size of formed QDs with this method. In the initial step for
QD synthesis, heating of the reaction solution may be
useful in order to begin particle growth. Therefore this
preparative approach based on the separation of nucleation
and growth very similar to the extremely successful TOP/
TOPO preparation of IIVI semiconductor QDs [44].
Nevertheless, second similarity between the two strategies
is the post-preparative treatment by using size-selective
precipitation to the crude cluster solutions [54].
For instance, Gaponik et al. [94] show a typical standard
aqueous synthesis of thiol-capped CdTe nanocrystals with a
naturally sulphur-capped surface (CdS shell) created by
mercapto-groups covalently attached to the surface
cadmium atoms (Fig. 5): 0.985 g (2.35 mmol) of
Cd(ClO4)2 6H2O is dissolved in 125 ml of water, and 5.7
Table 4 Lists some of the stabilising agents and precursor used for synthesis QDs and size of formed thiol-stabilised QDs
QDs
Size
Precursor
Stabilising agents
References
CdTe
1.32.4 nm
NA
Cd(ClO4)2 6H2O-NaHTe
Cd(ClO4)2 6H2O-H2Te
[94, 95, 99]

[73]
NA
5 0.5 nm
5.5 0.5 nm
1.42.2 nm
2.13.2 nm
1.33.9 nm
35 nm
CdCl2-NaHTe
Cd(ClO4)2 6H2ONaHTe
Cd(ClO4)2 6H2O-H2Te
Cd(ClO4)2 6H2O-NaHSe
2-mercaptoethanol, 1-thioglycerol
2-mercaptoethanol, 1-thioglycerol,
2-mercaptoethylamine (MA),
L-cysteine, 2-(dimethylamino)ethanethiol
mixture (1:1) of 1-thioglycerol and
2,3-dimercapto-1-propanol, TGA
3-mercaptopropionic
dithiol-functionalised ionic liquid (dTFIL)
mercaptopropionic acid
2-mercaptoethanol, 1-thioglycerol
TGA, thiolactic acid
1-thioglycerol
thioglycerol
CdSe
CdS
Cd(ClO4)2 6H2O-H2S
CdSO4Na2S2O3 5H2O

doi: 10.1049/iet-nbt.2012.0028
[77]
[100]
[96]
[93]
[92]
[43]
65
www.ietdl.org
Fig. 5 Schematic presentation of the synthesis of thiol-capped CdTe QDs

First stage: formation of CdTe precursors by introducing H2Te gas
Second stage: formation and growth of CdTe nanocrystals promoted by reux [94]
mmol of the thiol stabiliser are added under stirring, followed

by adjusting the pH to the appropriate values (depending on
the stabiliser nature) by dropwise addition of 1 M solution
of NaOH. The solution is placed in a three-necked ask
tted with a septum and valves and is purged by N2 for
about 30 min. Under stirring, H2Te gas is passed through
the solution together with a slow nitrogen ow. H2Te can
produce by the reaction of 0.2 g (0.46 mmol) of Al2Te3
lumps with 1520 ml of 0.5 M H2SO4 under N2
atmosphere. H2Te gas can be generated by at least two
different methods: (1) electrochemical reduction of Te
electrode in acid, Te0 + 2H+ + 2e H2Te; (2) by
chemical decomposition Al2Te3 powder or lumps, Al2Te3
+ 3H2SO4 3H2Te + Al2(SO4)3. Control of pH is very
important factor which strongly inuenced the PLQE of
thiol-capped QDs. Li et al. indicated signicantly improved
the quantum yields of CdTe QDs up to 4067% at lower
pH value (88.2) [101]. Rogach et al. demonstrate that
optimise pH (12) and molar ratio of thiol to Cd ions (1.3:1)
increases the room-temperature photoluminescence quantum
efciency of as-synthesised CdTe capped by TGA to values
of 4060%.
Vossmeyer et al. have synthesized CdS with aqueous route:
a solution of 1.97 g (4.70 mmol) of Cd (ClO4)2 6H2O and 1
ml (11.53 mmol) of 1-thioglycerol in 250 ml of water was
adjusted to pH 11.2 with 1 M NaOH. The particle size was
controlled by the quantity of H2S added as well as the
temperature and duration of further heating. Thus, sample
was prepared by the addition of 25 ml (1.12 mmol) of H2S
and stirring for 2 h at room temperature [92].
Rogach et al. in 1999 produce aqueous colloidal solutions
of CdSe nanoparticles: a solution of 1.97 g (4.70 mmol) of Cd
(ClO4)2 6H2O and 11.54 mmol of the stabiliser (RSH) in
250 ml of demineralised water was adjusted to pH 11.2
with 1 M NaOH. The solution was placed in a three-necked
ask tted with a septum and valves and was purged with
N2 gas for 30 min. Under vigorous stirring, 44 ml (2.2
mmol) of the freshly prepared oxygen-free 0.05 M NaHSe
solution was injected. Solutions were reuxed for different
times (up to 12 h), and aliquots were removed at regular
intervals (1530 min) with absorption spectra taken to
monitor the growth of the clusters [93]. The method of
size-selective precipitation [54] was used to separate the
samples of CdSe nanoparticles with different sizes and
narrow size distributions from crude solutions.
66
7.2.1 Effective parameters for the synthesis of QDs

with aqueous route [102]:
1. Effect of reactant concentration: The Cd:Te, Cd:S, Cd:Se
molar ratio used for QDs synthesis plays an important role in
the quality of the resultant QDs.
2. Effect of precursor pH: The pH of the precursor solution is
a key factor that inuences the QY as well as the QDs growth
rate. The pH of the precursor solution for synthesis QDs was
kept within 89 because for CdTe/CdS QDs pH higher or
lower than this range causes particle aggregation resulting
in the decrease of their uorescence property.
3. Effect of heating temperature: The structure of the QDs
surface is dependent on the reaction temperature. Faster
growth rate because of higher temperatures minimises the
formation of surface defects and thus develop the
uorescence properties of the QDs. One important benet
of aqueous synthesis is that it permits high-temperature
synthesis of QDs.
4. Effect of heating time: Heating time is directly
proportional to the size of the QDs. Longer heating time
formed larger QDs that emitted at higher wavelengths.
Aqueous method for synthesis of QDs used in biology
research has several advantages and disadvantages
compared with organometallic approach. In Table 5,
compare of aqueous synthesis against organometallic
approach and shown advantages and disadvantages of
aqueous synthesis compared with organometallic approach.
7.3
Biosynthesis of QDs
The organometallic method and water-based syntheses are

two successful routes for produce the highly uorescent
heavy metal chalcogenide-based QDs [103]. However, the
recent methods involve highly toxic chemicals, such as
TOP or TOPO, high temperatures and non-polar solvents in
the synthesis procedure and limited their applications in
clinical elds [104]. Thus, development of clean,
biocompatible, non-toxic and eco-friendly methods for QDs
synthesis has very importance [105]. More recently, the
biosynthesis of QDs has been developed and attracted
signicant attention. This method for QDs synthesis
involves the utilise of biological organisms to prepare QDs
with controlled size, shape, chemical composition and
doi: 10.1049/iet-nbt.2012.0028
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Table 5 Compare of aqueous synthesis against organometallic approach
Advantages
Disadvantages
1. low cost
1. QDs synthesised by this method do not possess the degree of

crystallinity of the organometallically prepared QDs. In
organometallic synthesis can produce QDs with high quantum
yields and the size distribution can be easily controlled by alerting
temperature or reaction time [55, 94]. QDs with narrower particle
size distribution is obtained with organometallic method [94]
2. aqueous synthesis of thiol-capped QDs can be carried out

equally efficiently on a vast scale
3. synthesised QDs with this method can be precipitated, washed
and reserved in the dry state under ambient conditions as long as 2
years being stable and dissolvable in water
4. ease and high reproducibility
5. the smaller QDs are obtained with aqueous approach [94]
6. this method is environment-friendly
7. the possibility of controlling the surface charge and other surface
properties of QDs synthesis with aqueous process simply by the
selection of the stabilising agent with suitable free functional
groups is absolutely important, particularly when water-soluble
nanocrystals are needed, for example, for fluorescent tagging
applications [55, 94]
8. QDs synthesised via the organometallic method require an extra
processing step to achieve biocompatibility but in aqueous
synthesis the ligands used during formation of QDs become the
eventual biocompatible surface ligands and do not need an
exchange process [97]
9. safe reactants used in synthesis of QDs
functionality [90]. The synthesis of QDs using

microorganism is a comparatively new and unexplored area
and is cost effective, non-toxic, reproducible and can be
achieved at ambient temperatures and pressure [106]. Heavy
metals for instance Cd, inuence the microbial population
by affecting their growth, morphology, damage the cell
membranes, alter enzymes specicity, disrupt cellular
functions, damage the structure of the DNA and nally
resulting in decreased biomass and variety [107, 108].
Microbial populations exposed to toxic concentrations of
Cd and heavy metals have developed four main tolerance
mechanism [109].
1. Modication of membrane transport systems involved in
initial accumulation, thus denying or reducing entry of
cadmium [110].
2. Intracellular or extracellular suppression by specic
binding to a biopolymer, generally the cell wall [111, 112].
Tolerance to heavy metals can also be achieved by
extracellular precipitation with microbially produced
oxalate, phosphate or sulphide [113115] or extracellular
polymers [116].
3. Energy-dependent ion efux [117].
4. Detoxication by enzymatic oxidation or reduction of the

ion to a less toxic form or covalent modication [117].
With mentioned mechanisms, microorganisms such as
bacteria and fungi can produce QDs when encounter with
Fig. 6 Photographs showin

a TEM and
b HRTEM images of extracellularly biosynthesised CdTe QDs [121]
Table 6 List of some microorganisms that synthesise CdS, CdTe and CdSe QDs
QDs type
CdS
CdTe
CdSe
Size
Microorganism/morphology
Localisation
References
520 nm
25 nm
22.5 nm
ND
8.01 0.25 nm
ND
5200 nm
20200 nm
30 nm
23 nm
23.6 nm
915 nm
F. oxysporum
E. coli
Schizosaccharomyces pombe
Candida glabrata
R. palustris
Clostridium thermoaceticum
Klebsiella pneumoniae
Klebsiella aerogenes
Gluconoacetobacter xylinus
E. coli
S. cerevisiae
F. oxysporum
extracellular/hexagonal
intracellular/spherical
intracellular/hexagonal
intracellular
intracellular/cubic
cell surface
cell surface
cell wall/spherical
cellulose bre/spherical
extracellular/spherical
extracellular/cubic zinc blende
intracellular/spherical
[118]
[120, 123]
[124, 125]
[124]
[119]
[126]
[127]
[128]
[129]
[121]
[103]
[106]

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Table 7 New methods for synthesis of bioapplicable QDs
QDs
CdS
Synthesis methods
solvothermal technique
non-injection one-pot method without
nucleation initiators
sono-micro-emulsion method
CdSe
CdSe/
CdS
CdSe/
Cdx Zn1
xS
CdSe
3 nm
4.2
0.2 nm
2 nm
one-pot non-injection hydrothermal
3.5 nm
high-intensity ultrasonic irradiation
6 nm
Elemental
sources-solvent-capping
ligand
S dissolved in ethylene
glycol, Cd(NO3)2
cadmium stearate, S
powder,
N-oleoylmorpholine
CdCl2, S, p-xylene,
ethylenediamine, CTAB,
1-butanol
CdCl2 2.5H2O, thiourea,
MPA
CdCl2, Na2SeSO3, MAA
aqueous synthesis
3.4 nm
CdCl2, H2Se, H2S
microemulsion-based technique
4.88 nm
Cd(NO3)2, Se, Na2SO3,

(NH4)2S, AOT, n-heptane
organic route
5.47.2
nm
Cd(AC)2 2H2O, S, Se, Zn

(AC)2, TOP, OA, TOA
high-intensity ultrasonic irradiation
13 nm
CdCl2, Na2SeSO3, MAA
green synthetic route

green synthetic route protein-directed
aqueous synthetic
CdTe
Particle
size
layer-by-layer method
5 nm
CdCl2, Se, BSA

2
4 nm
CdCl2, Se aqueous
solution, Lyz
2.3 nm
CdCl2 2.5H2O, SeO2,

NaBH4, TGA
CdCl2, NaHTe, TGA,
APTES
5 nm
photochemical synthesis
5.9 nm
CdCl2, NaHTe, TGA
aqueous synthesis
26 nm
CdCl2, NaHTe, L-cysteine
hydrothermal synthesis
3 nm
CdCl2, NaHTe, GSH, TGA
CA-CdTe
hydroxylamine-hydrochloride-promoted
4 nm
CdCl2, Na2TeO3, NaBH4,

NH2OH HCl, CA
CdTe/
CdSe
hydrothermal synthesis
4.8 nm
CdCl2 2.5H2O, Na2TeO3,

Na2SeO3, MPA
CdTe/
ZnS
aqueous synthesis
3.7 nm
CdCl2 2.5H2O, NaHTe,

ZnCl2, GSH
CdTe/
CdS
hydrothermal treatment method
3.5 nm
CdCl2 2.5H2O, Na2TeO3,

MSA, NaBH4, Na3C6H5O7
aqueous-phase synthesis
3 nm
heavy metals. Many microorganisms, such as Fusarium

oxysporum [118], Rhodopseudomonas palustris [119] have
been used to synthesise CdS QDs. Mi et al. [120] were
prepared CdS QDs in genetically engineered Escherichia
coli through the introduction of foreign genes encoding a
CdS-binding peptide. The CdS QDs were successfully
separated from the bacteria through two methods: lysis and
68
CdCl2 2.5H2O, NaHTe,

TP, TGA, GSH, TAA
Advantages
References
soluble CdS, highly stable,

stabilise without capping
agent
cost-effective, large-scale
production, green
[135]
simplicity, uniform shape,

synthesis at low
temperature
water-dispersible, high
crystallinity, high
photochemical stability
water soluble, increased
PLQY
low-toxic chemicals,
large-scale, water-soluble,
controllable introduction of
Se and S sources
simple, green, large-scale
synthesis, water-soluble,
bio-taggable
less toxic, insensitive to air,
easier to purify
[137]
water-soluble, easy control

of reaction conditions
water-soluble,
biologically-compatible
water-soluble, excellent
uorescent QDs, specically
biological function
less toxic, good stability
[142]
excellent molecules
recognition ability,
inexpensive, water-soluble,
environment-friendly
environment-friendly, high
photo stability
water-soluble, good
uorescent properties, short
synthesis period, the
broaden range of starting
pH value, less toxicity
good biological
compatibility, optical
stability
water-soluble, very simple
synthesis method, excellent
stability
water-soluble, excellent
photo stability, good
monodispersity
less toxicity, better
biocompatibility,
environmental friendly
low cytotoxicity, higher
chemical stability in a pH
range of 69, water-soluble
environmentally friendly,
highly biocompatible,
simple
[136]
[138]
[130]
[139]
[140]
[141]
[143]
[144]
[145]
[146]
[147]
[148]
[149]
[150]
[151]
[152]
[153]
[154]
freezingthawing of cells. In one study, Bao et al.

demonstrate a easy and efcient biosynthesis of highly
uorescent CdTe QDs with uniform size (2.03.6 nm) by
incubating a type of widely used and easily cultured
microorganism yeast cells (Saccharomyces cerevisiae)
with inexpensive inorganic Cd and Te salt precursors,
which has a high yield of nearly 90%. The extracellularly
doi: 10.1049/iet-nbt.2012.0028
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bio-grown CdTe QDs can be easily harvested and show
tunable size-dependent emission from 490 to 560 nm with a
relatively high photoluminescence quantum yield (PLQY)
of 33%. In addition, the CdTe QDs are naturally capped
with proteins in the biosystem, and show excellent water
solubility, great stability and biocompatibility for use in
biology eld [103]. Fig. 6a shown TEM image of the CdTe
QDs synthesised in E. coli (B) The high-resolution TEM
micrograph shown the size of the QDs is 23 nm.
Kumar et al. [106] showed that plant pathogenic fungus
F. oxysporum when challenged with aqueous CdCl2, SeCl4
at room temperature for 96 h under shaking conditions on a
rotary shaker (200 rpm) resulted in the formation of highly
stable CdSe nanoparticles possessing the following
characteristics: 11 nm, polydisperse, with broad
photoluminescence spectrum and 67 ns uorescence
half-life.
Microbial synthesis of QDs can be done either intracellular
or extracellular. The extracellular synthesis of semiconductor
nanoparticles makes it possible to harness and immobilise/
deposit such nanoparticles onto desired solid surfaces for
different practical purposes and has wider applications in
optoelectronics, bioimaging and in sensor technology than
intracellular accumulation. In the intracellular production,
the accumulated particles have less polydispersity. In order
to release the intracellularly synthesised QDs, extra
processing steps for example ultrasound treatment or
reaction with suitable detergents are required [106, 122].
Albeit biosynthesis of QDs are regarded as safe,
cost-effective, sustainable and eco-friendly processes, they
also have some disadvantages in culturing of microbes,
which is time-consuming and difcult in providing better
control over size distribution, shape and crystallinity. QDs
prepared by this method are also not monodispersible and
the rate of production is slow. These are the problems in
synthesis of QDs with microorganisms, but the information
obtained from strain selection, optimising the conditions
such as pH, incubation temperature and time, concentration
of metal ions, and the amount of biological material has
come up to give hope in achievement of these approaches

in large scale and for biomedical applications [122].
The microbial synthesis of QDs meets some challenges:
1. Improved control over size and shape
2. Scaling up of the synthesis to obtain large amount of QDs
3. Understanding the synthesis mechanism at the molecular
level, which nally may help in providing better control
over size, shapes and crystallinity in the future [120].
Table 6 shows some microorganisms that synthesise CdS,
CdTe and CdSe QDs.
8 Recent efforts to synthesise QDs for

biomedical applications
We have noted that QDs prepared by organometallic methods
are hydrophobic and need additional chemical modication to
make them water-soluble. This process is usually
time-consuming and causes a decrease in PL efciency.
QDs synthesised with aqueous and biosynthesis methods
have problems such as low crystallinity, wider particle size
distribution, low QY and time-consuming and therefore,
these methods should be improved. So, new methods for
the synthesis of appropriate QDs for biomedical
applications are needed. Bao et al. synthesised CdSe and
CdSe/CdS
QDs
in
aqueous
solution
with
ultrasound-assistance by using MAA as the capping ligand.
The PLQY of CdSe/CdS QDs were increased 3 times more
than that of the original CdSe by around 3 times [130]. A
group of researchers reported a protocol for biomimetic,
mild chemical synthesis of CdTeglutathione (GSH) QDs
in microorganisms with improved biocompatibility. They
have used CdCl2, k2TeO3 and GSH, and highly uorescent
QDs were synthesised when microorganisms grew. Toxicity
of CdTe QDs was lower than that observed with QDs
produced by other methods [131]. Recently, a new method
developed for the synthesis of CdS QDs through
micro-emulsion (O/W) induced by ultrasound. QDs with a
Fig. 7 Schematic presentation of steps involved in the bioconjugation of QDs [155]

doi: 10.1049/iet-nbt.2012.0028
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Table 8 More details for toxicity of QDs are shown in table [45, 160, 161]
QD
Model
Administration
QD
concentration
Exposure
duration
Toxicity
Reference
CdSe/ZnSSSA
EL-4 cells
1 106 cells/well
0.10.4 mg/ml
024 h
[162]
CdSe/ZnSSSA
EL-4 cells
200 l cell
suspension
injected (iv) into
mice
5 104 cells/ml
0.1 mg/ml QDs

per 5 107 cells
2 h to 7 days
cytotoxic: 0.1 mg/ml

altered cell growth;
most cells
non-viable at 0.4
mg/ml
no toxicity in mice
in vivo
12 M
12 h
vero, HeLa and

primary human
hepatocytes
100 l QDs/3
104 cells
00.4 mg/ml
24 h
rat
pheochromocytoma
cells, murine
microglial cells
primary rat
hepatocytes
1 105 cells/cm2
0.01100 g/ml
224 h
62.51000 g/ml
18 h
xenopus blastomeres
5 109 QDs/cell
(0.23 pmol/cell)
Days
mice
200-l tail vein

injection
1.53 nl of 2.3
M QDs
injected, 2.1
9
10 to 4.2 109
injected QDs/
cell
Injections;
180 nM QD,
20 pmol QD/g
animal weight
400600 nM
15 min cell
incubations,
1133 days in
vivo
4560 min
no signs of
localised necrosis at
the sites of
deposition
no effects on cell
growth
0.51.0 M
15 min
[168]
60 M QD/g
animal weight,
1 M and 20 nM
nal QD
concentration
100 l of
B16F10 cells
used for tail
vein injection,
2 105 to 4
105 cells
injected
0.24 mg/ml
not given
no effect on cell
growth,
development
mice showed no
noticeable ill effects
after imaging
46 h cell
incubation,
mice
sacriced at
16 h
no toxicity observed
in cells or mice
[170]
2h
0.4 mg/ml MUA/

SSAQD complexes
did not affect
viability of vero
cells
10 nM QD had
minimal impact on
cell survival
nearly completely
inhibited cell growth
even from the very
beginning
at 1 nM did not
initiate any
detrimental effects;
at 100 nM resulted
in the death of all
cells
[171]
CdSe/ZnS conjugates:
NH2, OH, OH/COOH,
H2/OH, MUA, COOH
CdSe/ZnSMUA
CdTe
CdSeMAA, TOPO
QDs
QD micelles: CdSe/
ZnS QDs in (PEGPE)
and
phosphatydilcholine
CdSe/ZnS amp-QDs
and mPEG QDs
CdSe/ZnSDHLA
avidin-conjugated
CdSe/ZnS QDs
CdSe/ZnS
amphiphilic micelle
WTK1 cells
dictyostelium
discoideum and HeLa
cells
HeLa cells
mice
tail vein injection
CdSe/ZnSDHLA QDs
mice, B16F10 cells
5 104 B16F10
cells with 10 l
QDs (10 pmol),
tail vein (iv)
injection
CdSe/ZnSMUA QDs;
QDSSA complexes
vero cells
0.4 mg/ml
CdSe/ZnS
HeLa cells
1 106 cells
HEK293 cells
PC12 cells
CdTe aqQDs
CdTe-gelatinised/
non-gelatinised
10 days (cell
culture)
1 105 cells
10 pmol QDs/
1 105 cells
(10 nM)
300 or 600 nM
1 105 cells/cm2
1 to 100 nM
72 h
3 days
[162]
2 M QDCOOH
induced DNA
damage at 2 h
cytotoxic: 0.2 mg/
ml, vero; 0.1 mg/ml,
HeLa; 0.1 mg/ml,
hepatocytes
10 g/ml cytotoxic
[163]
cytotoxic: 62.5 g/
ml cytotoxic under
oxidative/photolytic
conditions no
toxicity on addition
of ZnS cap
5 109 QDs/cell: cell
abnormalities,
altered viability and
motility No toxicity
at 2 109 QDs/cell
[166]
[164]
[165]
[167]
[24]
[168]
[169]
[172]
[173]
[174]
Continued
70

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Table 8 Continued
QD
Model
Administration
QD
concentration
Exposure
duration
Toxicity
Reference
K562 and HEK293T

human cell lines
1 105 cells
0.23.0 M
048 h
[175]
CdSe/ZnSPEG
(EviTag T1 490 QD)
caco-2 (human colon

carcinoma) cell line
106 cells/ml, 0.2

ml/well
0.84105 M
024 h
CdSe
primary rat
hippocampal neuron
cells in culture
104105 cells/ml
1, 10 and 20 nM
24 h
cells treated with

CdTe and CdTe/CdS
QDs were mostly
non-viable by 48 h
(for all
concentrations
tested)
commercially
available QD
demonstrated low
cytotoxicity but
induced cell
detachment
1 nM QD for 24 h
showed no
decrease in cell
viability; in contrast,
cells treated with 10
and 20 nM QD for
24 h showed
decreases in cell
viability of the order
of 20 and 30%
CdTe, CdTe/CdS,
CdTe/CdS/ZnS
hexagonal phase were prepared at a relatively low temperature

(70C), short time and fast transition phase. The main point of
this work is to achieve the CdS QDs in the presence of
ultrasound without additives and calcinations [132]. Zhao
et al. reported the preparation of a series of water-soluble
CdSe and CdSe/CdS QDs using amino acid-modied -CD
as surface coating agent by a simple sonochemical method.
These QDs are very soluble and stable in biological buffer,
and have appropriate size, high colloidal stability, good
biocompatibility, low toxicity and strong optical emission
properties [133]. Water-soluble CdSe QDs have been
prepared by a new tow-step method. First, H2Se was
produced by the reaction of the Na2SeSO3 solution with
diluted H2SO4, and then was ventilated into Cd2+ capped
with 3-MPA solution to form QDs nucleus; second, CdSe
solution was poured into a Teon-lined stainless steel
autoclave to obtain the desired CdSe QDs by controlling
the particles growth time and temperature. The results show
the water-soluble CdSe possess higher purity and PLQY,
improved photostability, and less synthetic time as
compared with aqueous CdSe QDs obtained by other
methods [134]. Table 7 summarised recent new methods for
synthesis of bioapplicable QDs.
9 Attachment of molecules on the surface of

QDs for targeting (bioconjugation)
Biocompatible QDs have to be cross-linked to biomolecules
such antibodies, oligonucleotides, nucleic acids or small
molecule ligands to render them specic to biological
targets and therefore be used for diagnosis and targeting
drug delivery. For bioconjugation, these molecules can be
attached either covalently or non-covalently on the surface
of QDs. QDs attached to small molecule ligands, inhibitors,
peptides or aptamers can bind with high specicity to many
different cellular receptors and targets. Functional ligands of
QDs surface such as carboxylic acids, primary amine and
thiol can be conjugated with antibodies and peptides by
exploiting cross-linking chemistry of carbodiimide,
doi: 10.1049/iet-nbt.2012.0028
[176]
[177]
maleimide
and
succinimide.
Also,
avidinbiotin
cross-linking is one of the most popular methods for
conjugating biomolecules on the surface of QDs [84, 87,
155]. These methods are shown in Fig. 7.
10
QD toxicity
Toxicity of QDs for biological applications is a critical factor.

There are different opinions about toxicity of QD. Yan et al.
shown that QDs could not only impair mitochondria but also
exert endothelial toxicity through activation of mitochondrial
death pathway and induction of endothelial apoptosis [45,
156]. Chen et al. results suggest that the cytotoxicity of
CdTe QDs not only comes from the release of Cd2+ ions
but also intracellular distribution of QDs in cells and the
associated nanoscale effects [157]. In other side, Ye et al.
shown that acute toxicity of these QDs in vivo can be
minimal. They demonstrated that rhesus macaques injected
with phospholipid micelle-encapsulated CdSe/CdS/ZnS
QDs, blood and biochemical markers remained within
normal ranges following treatment, and histology of major
organs after 90 days showed no abnormalities [158]. In the
other study, histological and biochemical analysis, and body
weight measurement demonstrate that there is no overt
toxicity of QDs in mice even at long-time exposure time
[159]. Table 8 demonstrated more results for toxicity of QDs.
11
Conclusion
In this review, we discussed three methods for synthesis of

heavy metal chalcogenide-based QDs for use in biomedical
eld. Also, we discussed new methods for synthesis of
bioapplicable QDs. We briey discussed the applications,
toxicity, photostability of QDs and surface engineering of
QDs for use in biology. The most popular method of QDs
synthesis is organometallic synthesis. The high reaction
temperature in this method ( >150350C) has two major
advantages: facilitates the removal of crystalline defects and
allows improvement in the photoluminescence. Size
71
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distribution of QDs prepared by this method can be easily
controlled by alerting temperature or reaction time. CdSe,
CdS or CdTe QDs synthesis with organometallic has a
comparatively low QY (brightness), usually less than 10%,
requiring a shell of a high band-gap semiconductor, such as
CdS or ZnS to be epitaxially grown around the core which
increases the QY up to 80%. This layer moreover protects
the core from oxidation and prevents leaching of the Cd/Se.
The main drawback of this technique is that the QDs
require the ligand exchange or encapsulation to make them
compatible in biological solutions. Direct aqueous synthesis
of QDs by applying different thiol groups as stabilising
agents provides an effective alternative to widely used
synthetic routes in highly boiling organic solvents.
Advantages of this method compared of organometallic are
ease and high reproducibility, low cost, vast scale and
synthesised QDs with this method can be precipitated,
washed and reserved in the dry state under ambient
conditions as long as two years being stable and dissolvable
in water and this method is environment-friendly. In this
method the ligands used during formation of QDs become
biocompatible surface ligands and do not need an exchange
process. The organometallic method and water-based
syntheses are two successful routes for produce the highly
uorescent heavy metal chalcogenide-based QDs. However,
the recent methods involve highly toxic chemicals, such as
TOP or TOPO, high temperatures and non-polar solvents in
the synthesis procedure and limited their applications in
biomedical elds. Albeit biosynthesis of QDs are regarded
as safe, cost-effective, sustainable and eco-friendly
processes, they also have some disadvantages in culturing
of microbes, which is time-consuming and difcult in
providing better control over size distribution, shape and
crystallinity. QDs prepared by this method are also not
monodispersible and the rate of production is slow. These
are the problems in synthesis of QDs with microorganisms,
but the information obtained from strain selection,
optimising the conditions like pH, incubation temperature
and time, concentration of metal ions, and the amount of
biological material has come up to give hope in
achievement of these approaches in large scale and for
biomedical applications. In recent years, scientists have
worked on novel methods for the synthesis of QDs with
suitable properties for use in medical. The new techniques
are often based on improved aqueous synthesis and
biosynthesis of QDs. These methods are often used CdCl2
as a source of cadmium and short chain thiols as stabilising
agents and they could synthesis QDs with advantages such
as low toxicity, better biocompatibility, and short synthesis
period, environmental friendly and improved photostability.
12
Future prospects
According to unique properties of QDs, they are suitable

alternatives to conventional organic uorophors for
labelling cells, investigating cellular process, and for many
other imaging applications. Although current synthesis
methods of QDs somewhat capable to prepare appropriate
QDs for utilise in biology, but to overcome the present
problems in QDs synthesis, in the near future, there are
several areas of research that are particularly promising and
should be paid enough attention to:
1. Improvement of aqueous route and synthesis QDs with
enhanced QY and narrower size distribution.
72
2. Changing the conditions of reaction media in synthesis of

QDs that cause to produce QDs with improved characteristics.
3. Use of safe solvent and stabiliser in QDs synthesis
methods.
4. Development of novel type of QDs without heavy metals
such as cadmium.
5. With changing the condition of reaction such as heating
temperature and heating time, pH, the amount of precursors,
reactant Concentration and can be produced QDs with
suitable properties for use in biology and medicine.
6. Future research on microbes-mediated biological synthesis
of QDS: using different species of microorganisms that live in
harsh conditions and changing the growing parameter in
culture media can produce improved QDs. In addition, this
method has some challenges such as scaling up of the
synthesis to obtain large amount of QDs, improved control
over size and shape and understanding the synthesis
mechanism at the molecular level, which nally may help
in providing better control over size, shapes and
crystallinity in the future.
13
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Review: three synthesis methods of CdX (X = Se,

IET Nanobiotechnol., 2014, Vol. 8, Iss. 2, pp. 5976

Interest in colloidal inorganic nanoparticles such as

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emission spectra, independence of emission on the

Fig. 1 Schematic drawing demonstrated

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IET Nanobiotechnol., 2014, Vol. 8, Iss. 2, pp. 5976

QDs are constructed from semiconducting materials. The

Design principles of QDs for biological uses

QDs modelling for biological use includes: (i) Synthesise the

QDs for use in eld of biology consist of:

organic solvents have hydrophobic surface ligands such as

The application of QDs, as a new technology for biosystems,

6 Stability of QDs in live cells and

Lattice parameter, [A]

IET Nanobiotechnol., 2014, Vol. 8, Iss. 2, pp. 5976

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Fig. 2 Demonstrated some of QDs applications in medicine and biology [45]

of environment have signicant impact on the stability of

In the early 1980s, several pioneering groups began to work on

Organometallic precursor synthesis of QDs

With entrance of nucleation and growth techniques to

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synthesis of QDs made easier and more controllable. An

Fig. 3 Schematic diagram showing

chalcogenide QDs, cadmium precursors are organometallics

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Precursors and stabilisers

CdMe2, diethylzinc (ZnEt2), TOP-Se, hexamethyldisilathiane

Cd(Ac)2 or CdO or CdCO3, TOP-Se, stearic acid or lauric acid

TOP-Se, CdO, Cd/oleic acid

to a Schlenk line (vacuum gas manifold) with one of the

aliphatic coordinating ligands (such TOPO) are only soluble

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IET Nanobiotechnol., 2014, Vol. 8, Iss. 2, pp. 5976

cubic CdSe (zinc blende)

cubic CdSe (zinc blende)

cubic CdSe (zinc blende)

these two strategies may be composed of charged moieties

Aqueous route for synthesis QDs

Typically, CdSe nanocrystal synthesis includes the use of Cd

the aqueous solution [98]. Using this method, CdSe

[94, 95, 99]

IET Nanobiotechnol., 2014, Vol. 8, Iss. 2, pp. 5976

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Fig. 5 Schematic presentation of the synthesis of thiol-capped CdTe QDs

mmol of the thiol stabiliser are added under stirring, followed

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7.2.1 Effective parameters for the synthesis of QDs

The organometallic method and water-based syntheses are

1. QDs synthesised by this method do not possess the degree of

2. aqueous synthesis of thiol-capped QDs can be carried out

functionality [90]. The synthesis of QDs using

4. Detoxication by enzymatic oxidation or reduction of the

Fig. 6 Photographs showin

IET Nanobiotechnol., 2014, Vol. 8, Iss. 2, pp. 5976