Lecture 36

20-4-18
Applications of optical sensors
 RECAP
• SERS (Surface enhanced raman spectroscopy)

• SPR (Surface plasmon resonance)

• Interferometry : Mac-Zehnder and Youngs

• Resonant mirror
 Applications of Optical Sensors

Optical sensors based on Acid-Base Indicators

Optical sensors for Acidic

Optical pH sensors
and Basic gases
 Optical pH sensors
• Make use of acid-base indicators that are organic
compounds
• Acid dissociation is accompanied by change in color
HIn(a1)In-(a2)+H+; Ki = [In-][H+]/HIn (In means indicator)
a Are activities and Ki is indicator constant (equilibrium
constant)

An absorbance-pH relation can be derived by:

ct = [In-]+ [HIn], assuming that only basic species absorbs
light and taking into pH definition,
pH = -log(aH+)  -log(H+)
[In-] = Ki ct/(10-pH+Ki)
Ct is total concentration of indicator
• Plot of [In-] vs pH gives S-shaped curve

Possible to broaden the pH range by incorporating in the

sensing layer several pH indicators with pKi value selected so
as to give an absorbance superposition on extended and
pseudolinear response region.

Indicator constant depends on strength of the solution and

temperature (these two parameters need to be kept under
control)

Acid-base indicators can be immobilized by covalent bonding

or entrapment in cellulose membranes or polymeric materials

Sol-gel chemistry: indicator immobilization by entrapment

• pH indicators are covalently immobilized on
controlled-pore glass beads attached to the end of
the plastic optical fiber by heating.
• Thymol blue (pH range 1.2-2.8)
• Bromophenol blue ( 3-4.6)
• Common fluorescent pH indicators:
Fluorescein, Fluorescein amine, 8-Hydroxypyrene-
1,3,6-trisulfonic acid
Ratiometric method: pH = pKi + log[ln-]/Hln]
(Usually implemented in pH sensors when both acid
and base form of the indicator absorb or fluoresce at
specific wavelengths)
Fluorescein Fluorescein amine

8-Hydroxypyrene-1,3,6-trisulfonic acid
 Optical sensors for acid-base gases
For example: CO2 acidic gas NH3 basic gas
• Impart dissolution in water and effect pH , depends on the
concentration of gas sample
• Potentiometric method: pH change monitored by pH glass
electrode

• The indicator dye should be selected so as to match its

indicator constant to the ionization constant of the
dissolved gas

• pH buffer should be incorporated in the sensing layer in

order to prevent complete dissociation of indicator, this
would limit the response range of the sensor
• In clinical chemistry: The sensing layer in an
optical CO2 sensor is composed of an aqueous
solution of hydrogen carbonate as the pH buffer
and a light absorbing indicator dye. The solution is
entrapped at the end of the optical fiber bundle
by means of  10 m thick Teflon membrane that
is permeable to only CO2 but not to ions.

• Ammonia sensor has been developed using

chlorophenol red and bromothymol blue. About
0.2 to 30M ammonia can be sensed
• In fluorescence based gas sensors for ex: CO2 sensor
has been developed using the HPTS fluorescent
indicator incorporated in a silicone matrix as an
emulsion of a room temperature ionic –liquid. The
response function of this sensor can be put in a
linear form as
F0/F = 1+kcCO2
F0 and F are intensities in the absence and presence of
CO2

k is the constant depending upon the first dissociation

constant of carbonic acid, indicator constant and total
concentration of bicarbonate in the buffer solution
incorporated in the sensing film.
 Optical oxygen sensors
• The property of oxygen to produce dynamic fluorescence
quenching forms the basis of oxygen optical sensors

• Common fluorophores used in oxygen sensors are

ruthenium(II) complexes with 2,2‘-bipyridine(bipyridyl),
1,10, phenanthroline and related compounds. Also,
analogue complexes of platinum group metals as well as
certain organic dyes and polyaromatic hydrocarbons have
also been used
 Optical enzymatic sensors
• Can be obtained by combining an enzyme
layer with an optical system in order to
monitor reactants or products involved in
the enzymatic reactions.
• Depending upon the mechanism of the
enzymatic reaction either hydrogen ions,
oxygen, hydrogen peroxide, certain gases
or ammonium ion can serve for
transduction by means of optical probes.
 Optical Affinity sensors
• Luminescence is most widely used to perform
transduction in immunosensors
• FRET is method of choice in optical
immunosensors
• Label-free transduction method gives promising
results.
• Affinity reactions based on natural receptors
allows developing optical sensors for small
molecules such as glucose.
• Used in in-vivo applications
 Optical DNA sensors and Arrays
• Fluorescence labels can be appended to
nucleic acids in two ways :

Covalent bonding
Intercalation

• Cyanine dyes are common fluorescent

labels used in optical nucleic acid essays
• An alternate method is based on fluorescent intercalating
and groove binding of planar molecule dyes.

• When such a label is present in an aqueous solution,

fluorescence is quenched by water molecules. After
intercalation in a double-strand nucleic acid, fluorescence
is strongly enhanced owing to hydrophobic environment
between the base pairs.

Most common labels are

ethidium bromide and
thiazole orange dyes

Ethidium bromide
Thiazole orange
Nanomaterial application in
Optical Transduction
• Nanomaterials possess outstanding photophysical
properties such as semiconductor nanocrystals,
carbon nanotubes and metal nanoparticles

• Superior to traditional molecular luminophores and

are expected to replace them in many aspects

• Using optical waveguides on nanoparticles one can

develop chemical sensors
 Nanomaterial applications in optical
Transduction

Quantum dots
Lanthanides
Carbon nanotubes Porous silicon
Metal nanoparticles

 Note: Quantum dots, Carbon nanotubes, Metal nanoparticles and silica nanoparticles were
already studied in previous lectures hence we will study the remaining two of the above i.e.
 Porous silica and Lanthanides
 Luminescent lanthanide compounds
as nanomaterials
• Gd2O3 doped with Tb or Eu are highly
photostable, characterized by narrow emission
bands, have long life time and possess large
stokes shift.

• Polysiloxane
• Polysiloxane shell has been used for covering
Tb:Gd2O3 nanoparticles
• Helps in functionalization of surface and Tb-OH
group is removed.
• Eu-Gd2O3 nanoparticles covered with poly (L-
lysine) have been used as reporter labels-
biomarkers.

Eu3+ and Tb3+micro and nano

sized particles
 Porous Silicon
• It’s a form of chemical element silicon obtained by
chemical, electrochemical or photochemical etching of
nanocrystalline silicon in the presence of Hydrofluoric acid

• The structure is composed of aligned pores and silicon

columns perpendicular to the surface. Si-H surface react
with water, oxygen or other oxidizing agents, thus oxygen
is incorporated in the lattice forming various oxygen
functionalities.

• OH helps in the functionalization by condensation reaction

with chlorosilanes or alkoxysilanes.
• Porous silicon allows optical transduction by luminescent
quenching

• This property makes them use as gas sensors,

immunosensors or nucleic acid sensors

• Produces light reflection at the upper surface and at the

bulk silicon surface  differ in path length  interference
phenomenon pattern

• Binding of the analyte leads to the corresponding change in

the thickness which produces a shift in the interference
fringes, depending on the concentration of the analyte
 Metal nanoparticles in optical sensing

• Colloidal solutions of gold and silver have bright color

when light passes through it

• Fluorescence is the transition between the valence

orbitals in molecules

• In case of colloidal solution of metals, light excitation and

scattering is caused by the collective oscillation.
• Spherical nanoparticles behave as an oscillating dipole,
become like a light source that is why light is scattered
(increased absorbance over a specific spectral region)
bright appearance

• Nonspherical nanoparticles (nanorods) show more than

one absorption band due to surface plasmons created
along various directions are energetically different

• Thus an oblate nanoparticles supports two oscillation

modes longitudinal and transverse

Length of nanorods can be adjusted by aspect ratio i.e. L/R