Optical sensing

MSc. Lab on a chip – 2024

Dr.ir. J.E. (Nienke) van Dongen
[email protected]

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Why talk about sensors in a LoC course?
• Sensors are the “eyes and ears” of LoC devices
• Analyte detection
• Data collection
• Control
• Sensors for LoC have special requirements:
• Miniaturization
• Sensitivity & selectivity
• Cost-effectiveness
• Integration (or not?)

https://www.gene-quantification.de/lab-on-chip.html

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 Why me?
• PhD in optical biosensing
• Working on optical biosensors for
early cancer detection.

Gold nanoparticles CRISPR/Cas 3

 In this lecture:

1. 2. 3. 4. 5. 6. 7.
Learning outcomes Absorbance Luminescence Refractive index On-chip or off-chip? Wrap-up Assignments
Learning outcomes Optical sensing
At the end of these sessions, you are able to:
• Explain the operational principle of various optical detection methods
• Absorbance
• Luminescence
• Refractive index
• For a given experimental procedure:

• Design a LoC system with the necessary components for optical read-out

• Discuss the preferred integration levels of these components

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Types of optical sensing

https://www.sciencedirect.com/science/article/pii/S0003267007014444 6
Absorbance
• Absorbance is the quantity of light
absorbed by a material/solution.
• Transmittance is the quantity of light
that passes through a solution.
• Absorbance and % transmittance are
molecule/material characteristics

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Absorbance to determine a concentration
Absorbance Equation:
A = -log10(I/I₀) = ε * c * l"
A: Absorbance
I: Intensity of transmitted light
I₀: Intensity of incident light
ε: Molar absorptivity (substance and wavelength dependent)
c: Concentration of the absorbing substance
l: Path length through the sample

https://www.thoughtco.com/beers-law-definition-and-equation-608172 8
 Microfluidics: size matters
1 cm 1 cm

• Less volume
• Less molecules
• Shorter path length

A = -log10(I/I₀) = ε * c * L
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Methods to increase the pathlength

DOI: https://doi.org/10.29363/nanoge.eimc.2021.021

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Luminescence

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Luminescence
“Emission of light (electromagnetic
waves) from a substance that does not
arise from heating.”

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https://www.edinst.com/blog/photoluminescence-differences/
 Photoluminescence: fluorescence
• 3-stage process Vibrational
• Excitation (By absorbance of photons) relaxation

• Excited-state lifetime (losing energy via vibrational relaxation)

• Fluorescence emission
Vibrational
relaxation
Vibrational
relaxation

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https://www.caltagmedsystems.co.uk/qrettech/
 Photoluminesence: phosphoresence
Immediate
• 3-stage process (10 -9 - 10 -7 s)

• Excitation Longer
Time-period
• Excited-state lifetime (10 -4 s - hours)

• Intersystem crossing (phosphorescence)

• Phosphorescence emission

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 Förster resonance energy transfer (FRET)
• If there is spectra overlap between
donor and acceptor
• Quenches the fluorescence
intensity of the donor while
increasing the emission intensity
of the acceptor.

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https://www.sciencedirect.com/science/article/pii/S0959440X15000913
 Förster resonance energy transfer (FRET)
• Scales with distance between
donor and acceptor
• Molecular Ruler

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https://www.nature.com/articles/srep33257 https://www.sciencedirect.com/science/article/pii/S0959440X15000913
 Chemiluminescence
Noctiluca scintillans

Electromagnetic wave,
heat, friction, electric field, Luminescence
chemical reaction, etc.

https://www.nationalgeographic.nl/natuur-leefomgeving/a44125909/zeevonk

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Chemiluminescence

Parvathi, A., et al. "Bioluminescence of green Noctiluca scintillans from the coastal waters of Kochi
(southwest coast of India) and in vivo experiments on bioluminescence in response to mechanical stimuli
and its diurnal variations." Marine Biology Research 17.7-8 (2021): 554-568.
 Chemiluminescence

https://www.sciencedirect.com/science/article/pii/S0956566318
306651

https://pubs.acs.org/doi/10.1021/acscentsci.2c01467

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 A

Lens

C
https://pubs.aip.org/aip/bmf/article/14/2/024109/134175/Optofluidic-
detection-setup-for-multi-parametric

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A

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Refractive index

“a value calculated from the ratio of

the speed of light in a vacuum to
that in a second medium of greater
density.”

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 Total internal reflection (TIR) sensing

• TIR occurs if θincidence > θcritical

• TIR depends on differences in
r.i. between two media Less dense
medium
• Can be used to measure binding

Dense
medium

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Formation of the Evanescent field
As the light wave "bounces" off the interface, it
creates a rapidly oscillating electromagnetic
field that extends into the less dense medium.
This field is known as the evanescent field.
Characteristics :
1. Decaying Intensity: The intensity of the
evanescent field decreases exponentially with
distance from the interface.
2. Limited Penetration: The evanescent field
penetrates only a short distance into the less
dense medium. It can extend from a few
nanometers to several wavelengths of light,
depending on the angle of incidence and the
wavelength of light. (250-5000 nm)
3. Non-Propagating: Unlike the incident and
reflected beams, the evanescent field does not
propagate through space as a wave but rather
exists as a near-field phenomenon.

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Biosensing using evanescent field
• Mach Zehnder interferometer
• Phase difference Δφ between
the sensing arm and the
reference arm
• Changes in refractive index
• intensity modulation caused by
the interference of the two arms
at the waveguide output.

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Surface plasmon resonance (SPR)
• relies on total internal reflection (TIR) at the metal-dielectric
interface.
• When the incident angle matches the resonance angle, surface
plasmons are excited, causing a reduction in reflected light intensity.
• The resonance angle is highly sensitive to changes in the refractive
index at the metal surface, making SPR a powerful sensing technique.
• Plasma field reaches till 250-1000 nm

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Surface plasmon resonance: measuring r.i.

Plasmon Wave

SPR reflection intensity (a.u.)

Prism

SPR angle

Dip Angle (degrees)

Max. absorption of light

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Surface plasmon resonance: measuring r.i.

SPR reflection intensity (a.u.)

Prism

SPR angle

Dip Angle (degrees)

Max. absorption of light

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Localized SPR
“optical phenomenon resulting from the interaction between light and
the conductive nanoparticles whose size is smaller than the incident
wavelength”
• No need for prism -> easier setup
• Plasma field 20 – 40 nm

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https://analytical-online.com/knowledge/comparison-between-lspr-spr
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Integrating optical sensors on chip

Advantages Disadvantages
• Miniaturization • Fabrication complexity
• High speed • Limited material choices
• Low crosstalk • High costs
• Re-usability

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Wrap-up
At after this session, you are now able to:
• Explain the operational principle of various optical detection methods
• Absorbance
• Luminescence
• Refractive index
• For a given experimental procedure:

• Design a LoC system with the necessary components for optical read-out

• Discuss the preferred integration levels of these components

37
 In this lecture:

1. 2. 3. 4. 5. 6. 7.
Learning outcomes Absorbance Luminescence Refractive index On-chip or off-chip? Wrap-up Assignments
Assignments
• What is up next?

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