Saliterman, Fundamentals of BioMEMS and Medical Microdevices, Ch.

10

Detection &
Measurement Methods
Introduction to BioMEMS
MN-BIO4600 Lecture 10
2015
 Detection and Measurement

Choosing a detection and measurement system for a specific

application may depend on the preference for a labeled
versus a label-free methodology

Some based on sensor principles found in clinical laboratory

medicine (Ch.8) suitable for integration in a microfluidic chip

Various measurement systems are also available for

characterization of Si, glass and polymer materials after
fabrication and/or surface modification
Detection Schemes
 Electrochemical Detection (EC)
Chemiluminescence
Fluorescence
Molecular Beacons
Aptamers
 See also chapter 6:
Electrochemical Detection Sensor Principles
& Microsensors

Potentiometry
measures the potential of a solution between two electrodes,
related to the concentration of one or more analytes.

Voltammetry (Amperometry)
Measures the current from a redox reactions as function of the
electrode potential. Typically consist of a three electrode
system.

Coulometry
determines the amount of matter transformed during an
electrolysis reaction by measuring the amount of electricity (in
coulombs) consumed or produced.

Conductometry
measures the electrical conductance of an electrolyte solution.
Unspesific (all ions in solution contribute to the conductance).
 Integrated CE and Electrochemical Detection

Electrochemicalanalysisinliquid
solutions
Signalbasedonelectricalquantities
(potential,current,charge)
Detectcompositionofsolution/
reactionkineticsofcontents/
products
Lessexpensivethanfluorescent
techniques
Electrodesintegratedaspartof
fabricationprotocol.
Portable:threeelectrode
potentiostat,9Vbattery
Electrodes:platinum0.3mthick,2.2
mmwide,6.9mmlong.
Channel:20mdeepx50mwide,1
cmlong.

Fig 1: Capillary electrophoresis with electrochemical detection chip.

 See also chapter 8:
Chemiluminescence Clinical Laboratory Medicine

Fig 2: Localization of HPV 18 nucleic acids (virus)

in a tissue section by chemiluminescence.
Luminol Chemiluminescence
Peroxyoxalate Chemiluminescence
 Fluorescence

Fig 3: Natural occuring fluorescent minerals.

Fluorescent Microscopy

Fig 4: Confocal Scanning Laser Microscopy.

Example: Multiple Exposure

Fig 5: Triple Exposure of a Drosophila Egg Chamber.

Fluorescent Microscopy: Confocal Microscopy

Opticalsectioningofcellsand
tissuesin3Dimages
Clearerandsharperimagesthan
conventionalfluorescent
microscopy
Colours usedintheseimages
representsspecificfluorescent
molecules
Excitedbyascanninglaserlight
beam

Fig 6: Confocal image of a human macrophage.

 Molecular Beacons

Singlestrandedoligonucleotide WhentheMBhybridizetothe
hybridizationprobesthatforma appropriatetarget,theprobe
stemandloopstructure. transitsfromdarktofluorescent.

Fig 7: Molecular beacons.

Molecular Beacons

Fig 8: Molecular Beacons

 Latinaptus fit&Greekmeros part)
Aptamers oligonucleotide orpeptidemolecules
thatbindtoaspecifictargetmolecule
SynthesizedbytheSELEXmethod
(SystematicEvolutionofLigands by
Exponentialenrichment)
chemicalequivalentofantibodies
Bindstotargetsbyvander Waals
forces,hydrogenbondingand
electrostaticinteraction
highlyspecific,relativelysmallinsize,
andnonimmunogenic
Oligonucleotide:
smallRNA/DNAmolecules(1560bp)
bindusingtheirentiresequenceto
proteinsorothercellulartargets
Peptide:
peptideswithasinglevariableloop
regiontiedtoaproteinonbothends
bindtotheirtargetswiththisvariable
loopregion.
Applications:
Therapeutics,Targetvalidation(amino
Fig 9: Biomolecule aptamer interaction. acid,protein),Drugscreening,Affinity
separation,Diagnosticsandbiosensors
Fig 9: The SELEX process (Systematic Evolution of Ligands by Exponential enrichment)
Measurement Systems
 Confocal Laser Microscopy
Interferometry
Ellipsometry
Surface Plasmon Resonance
Spectroscopy
Raman Microscopy (RS & SERRS)
Electron Microscopy (TEM & SEM)
Atomic Force Microscopy
X-ray tomographic microscropy
 Fluorescent Microscopy

Fig 10: Rotary Confocal Fluorescence Scanner used to interrogate radial micro capillary array
electrophoresis devices.
 Interferometry

Fig 11: Interference and diffraction.

 Interferometry

Fig 12: Michelson Interferometer.

 Interferometry

Fig 22: Surface profile from a white light interferometer

 Ellipsometry

Fig 13: Principle of operation of an ellipsometer.

 Surface Plasmon Resonance

Surfaceplasmon (polaritons)
formedalongthe
metal/dielectricinterface
Oscillateatoptical
frequencies
Monitoredbylightintensity
reflectedfromthe
dielectric/metalinterface
Minimumintensityatthe
resonanceofthesurface
polariton (plasmon)
Positionofbandsensitiveto
interfaceproperties(film
thickness,surfaceabsorbed
species,liquiddielectric
properties)
Anglechange 0.1 /nm

Fig 14: Surface Plasmon Resonance.

 Raman Microscopy (RS & SERRS)
Scatteringoflaserlight(nearUV near
IR)fromamaterial

Atinyfractionisshiftedinfrequency
duetovibratingatomsinthematerial

Analysisofthefrequencyshift
(spectrum)revealsthecharacteristic
vibrationfrequenciesoftheatoms
hencechemicalcompositionand
structureofmaterial

Particlesof1mcanbeidentified

Fig 15: Raman Microscopy.

Raman Microscopy (RS & SERRS)

Fig 16: Raman Microscopy.

Raman Microscopy (RS & SERRS)

phonon inc scat

Fig 16: Raman Microscopy.

Electron Microscopy (TEM & SEM)

Fig 17: SEM objects

Scanning Electron Microscopy (SEM)

Fig 18: SEM microscope

Transmission Electron Microscopy
(TEM)

Fig 19: TEM microscope

Atomic Force Microscope

Fig 20: AFM

Atomic Force Microscope

Fig 21: AFM

 X-ray tomographic microscropy

Fig 23: Synchrotron X-ray tomographic microscopy, The Swiss Light Source at Villigen, Switzerland, (2.4
GeV ) Paul Scherrer Institute, (Ref: Attenbourgh Origins of Life)
High resolution 3D imaging < 1m

Fig 24: Fossile embryos Markuelia hunanensis (540 480 mill BC).
 Anatomy of M.hunanensis

Fig 25: Image of embryo, revealing mouth with teeth, digestive tract and anus.
Summary - Detection & Measurement Methods

Detection Schemes:
Electrochemical Detection (EC)
Chemiluminescence
Fluorescence
Molecular Beacons
Aptamers

Measurement Systems:
Confocal Laser Microscopy
Interferometry
Ellipsometry
Surface Plasmon Resonance
Spectroscopy
Raman Microscopy (RS & SERRS)
Electron Microscopy (TEM & SEM)
Atomic Force Microscopy
X-ray tomographic microscropy
 Summary

Biomedical Micro Electro-Mechanical Systems

Topics of study (curriculum):
Introduction to BioMEMS
Principles of Biochemistry
Silicon and Soft Fabrication Techniques
Polymer Materials
Microfluidic Principles
Sensor Principles and Microsensors
Microactuators and Drug Delivery
Clinical Laboratory Medicine
Micro-Total-Analysis Systems
Detection and Measurement Methods
Genomics and DNA Microarrays
Proteomics and Protein Microarrays
Emerging BioMEMS technologies
Packaging, Power, Data, and RF Safety
Biocompatibility, FDA and ISO 10993
Thank you