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SECOND EDITION

Biomedical
Photonics
Handbook
Volume II
Biomedical Diagnostics
Biomedical Photonics Handbook, Second Edition

Volume I: Fundamentals, Devices, and Techniques

Volume II: Biomedical Diagnostics

Volume III: Therapeutics and Advanced Biophotonics

SECOND EDITION

Biomedical
Photonics
Handbook
Volume II
Biomedical Diagnostics

Edited by

Tuan Vo-Dinh
Duke University
Durham, North Carolina, USA
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 Inspired by the love and
infinite patience of
my wife, Kim-Chi, and
my daughter, Jade

This book is dedicated to the

memory of my parents,
Vo Dinh Kinh and Dang Thi Dinh
 Contents

Preface....................................................................................................................... ix
Acknowledgments.................................................................................................. xiii
Editor........................................................................................................................ xv
Contributors...........................................................................................................xvii
MATLAB Statement����������������������������������������������������������������������������������������������� xxi

SECTION I  Biomedical Analysis, Sensing, and Imaging

1 Biosensors for Medical Applications................................................................................. 3
Tuan Vo-Dinh, Leonardo Allain, and Andrew Fales
2 Glucose Monitoring........................................................................................................... 47
Casey W. Pirnstill, Brian Cummins, Gerard L. Coté, and Roger J. McNichols
3 Biochips and Microarrays: Tools for New Medicine. . .................................................... 77
Tuan Vo-Dinh and Guy D. Griffin
4 Atomic Spectrometry in Clinical and Biological Analysis......................................... 125
Andrew Taylor
5 Flow Cytometry. . ...............................................................................................................141
Francis Mandy, Joe Trotter, Rudi Varro, Ming Yan, and Diether Recktenwald
6 Capillary Electrophoresis Techniques in Biomedical Analysis...................................169
S. Douglass Gilman and Michael J. Sepaniak
7 Surface Plasmon Resonance Imaging Sensors. . ............................................................ 199
Julien Moreau, Jean-Pierre Cloarec, Paul Charette, Michel Goossens,
Michael Canva, and Tuan Vo-Dinh
8 Surface-Enhanced Raman Scattering for Biomedical Diagnostics............................ 265
Tuan Vo-Dinh and David L. Stokes
9 Functional Imaging with Diffusing Light...................................................................... 311
Arjun G. Yodh and David A. Boas
10 Molecular Contrast Optical Coherence Tomography ..........................................357
Oscar Carrasco-Zevallos and Joseph A. Izatt

vii
viii Contents

11 Multiscale Photoacoustic Microscopy and Macroscopy . . ................................... 383

Song Hu and Lihong V. Wang
12 Polarized Light for Medical Diagnostics ................................................................ 399
Steven L. Jacques
13 X-Ray Diagnostic Techniques .....................................................................................415
Xizeng Wu, Molly Donovan Wong, Abby E. Deans, and Hong Liu
14 Optical Pumping and MRI of Hyperpolarized Spins . . ........................................ 453
Xizeng Wu, Thomas Nishino, and Hong Liu

SECTION II  Biomedical Diagnostics and Optical Biopsy

15 Fluorescence Spectroscopy for Biomedical Diagnostics.. ................................... 485
Tuan Vo-Dinh and Brian M. Cullum
16 Elastic-Scattering Spectroscopy and Diffuse Ref lectance ................................. 543
Judith R. Mourant and Irving J. Bigio
17 Quantitative Diffuse Ref lectance Imaging of Tumor Margins ........................ 565
Bing Yu and Nirmala Ramanujam
18 Near-Infrared Fluorescence Imaging and Spectroscopy in Random
Media and Tissues . . ....................................................................................................... 587
E.M. Sevick-Muraca, E. Kuwana, A. Godavarty, J.P. Houston, A.B. Thompson, and R. Roy
19 Interferometric Light Scattering Techniques for Measuring Nuclear
Morphology and Detecting Dysplasia ......................................................................653
Yizheng Zhu, Francisco E. Robles, Neil G. Terry, and Adam Wax
20 Ultrasonically Modulated Optical Imaging . . ......................................................... 683
François Ramaz, Emmanuel Bossy, Michel Gross, and A. Claude Boccara
21 Optoacoustic Tomography: From Fundamentals to Diagnostic Imaging
of Breast Cancer .. ............................................................................................................715
Alexander A. Oraevsky
22 Raman Spectroscopy: From Benchtop to Bedside . . ...............................................759
Anita Mahadevan-Jansen, Chetan A. Patil, and Isaac J. Pence
23 Recent Developments in Fourier Transform Infrared (FTIR)
Microspectroscopic Methods for Biomedical Analyses: From
Single-Point Detection to Two-Dimensional Imaging . . ...................................... 803
Rohit Bhargava and Ira W. Levin
24 Diffuse Optical Spectroscopy: Frequency-Domain Techniques .......................819
Albert E. Cerussi and Bruce J. Tromberg
 Preface

In the tradition of the Biomedical Photonics Handbook, the second edition is intended to serve as an
authoritative reference source for a broad audience involved in the research, teaching, learning, and
practice of medical technologies. Biomedical photonics is defined as the science that harnesses light and
other forms of radiant energy to provide the solution of problems arising in medicine and biology. This
research field has recently experienced an explosive growth due to its noninvasive or minimally invasive
nature and the cost-effectiveness of photonic modalities in medical diagnostics and therapy.
The field of biomedical photonics did not emerge as a well-defined, single research discipline like
chemistry, physics, or biology. Its development and growth have been shaped by the convergence of
three scientific and technological revolutions of the twentieth century: the quantum theory revolution,
the technology revolution, and the genomics revolution.
The quantum theory of atomic phenomena provides a fundamental framework for molecular bio­
logy and genetics because of its unique understanding of electrons, atoms, molecules, and light itself.
Out of this new scientific framework emerged the discovery of the structure of DNA, the molecular
nature of cell machinery, and the genetic cause of diseases, all of which form the basis of molecular
medicine. The formulation of quantum theory not only gave birth to the field of molecular spectros-
copy but also led to the development of a powerful set of photonics tools—lasers, scanning tunneling
microscopes, and near-field nanoprobes—for exploring nature and understanding the cause of disease
at the fundamental level.
Advances in technology also played, and continue to play, an essential role in the development of
biomedical photonics. The invention of the laser was an important milestone. Laser is now the light
source most widely used to excite tissues for disease diagnosis as well as to irradiate tumors for tissue
removal in interventional surgery (optical scalpels). The microchip is another important technological
development that has significantly accelerated the evolution of biomedical photonics. While the laser
has provided a new technology for excitation, the miniaturization and mass production of integrated
circuits, sensor devices, and their associated electronic circuitry made possible the development of the
microchip, which has radically transformed the ways detection and imaging of molecules, tissues, and
organs can be performed in vivo and ex vivo. Recently, nanotechnology, which involves research on
materials and species at length scales between 1 and 100 nm, has been revolutionizing important areas
in biomedical photonics, especially diagnostics and therapy at the molecular and cellular level. The
combination of photonics and nanotechnology has already led to a new generation of devices for prob-
ing the cell machinery and elucidating intimate life processes occurring at the molecular level that were
heretofore invisible to human inquiry. This will open the possibility of detecting and manipulating
atoms and molecules using nanodevices, which have the potential for a wide variety of medical uses at
the cellular level. The marriage of electronics, biomaterials, and photonics is expected to revolutionize
many areas of medicine in the twenty-first century.

ix
x Preface

A wide variety of biomedical photonic technologies have already been developed for clinical moni-
toring of early disease states or physiological parameters such as blood pressure, blood chemistry, pH,
temperature, and the presence of pathological organisms or biochemical species of clinical importance.
Advanced optical concepts using various spectroscopic modalities (e.g., fluorescence, scattering, reflec-
tion, and optical coherence tomography) are emerging in the important area of functional imaging.
Many photonic technologies originally developed for other applications (e.g., lasers and sensor systems
in defense, energy, and aerospace) have now found important uses in medical applications. From the
brain to the sinuses to the abdomen, precision navigation and tracking techniques are critical to posi-
tion medical instruments precisely within the three-dimensional surgical space. For instance, optical
stereotactic systems are being developed for brain surgery, and flexible micronavigation devices are
being engineered for medical laser ablation treatments.
With the completion of the sequencing of the human genome, one of the greatest impacts of genomics
and proteomics is the establishment of an entirely new approach to biomedical research. With whole-
genome sequences and new automated, high-throughput systems, photonic technologies such as bio-
chips and microarrays can address biological and medical problems systematically and on a large scale
in a massively parallel manner. They provide the tools to study how tens of thousands of genes and
proteins work together in interconnected networks to orchestrate the chemistry of life. Specific genes
have been deciphered and linked to numerous diseases and disorders, including breast cancer, muscle
disease, deafness, and blindness. Furthermore, advanced biophotonics has contributed dramatically to
the field of diagnostics, therapy, and drug discovery in the postgenomic area. Genomics and proteomics
present the drug discovery community with a wealth of new potential targets. Biomedical photonics
can provide tools capable of identifying specific subsets of genes encoded within the human genome
that can cause the development of diseases. Photonic techniques based on molecular probes are being
developed to identify the molecular alterations that distinguish a diseased cell from a normal cell. Such
technologies will ultimately aid in characterizing and predicting the pathologic behavior of that dis-
eased cell, as well as the cell’s responsiveness to drug treatment. Information from the human genome
project will one day make personal, molecular medicine an exciting reality.
The second edition of this handbook is intended to present the most recent scientific and techno-
logical advances in biomedical photonics, as well as their practical applications, in a single source. The
three-volume handbook represents the collective work of over 150 scientists, engineers, and clinicians.
It includes many new topics and chapters such as fiber-optics probes design, laser and optical radiation
safety, photothermal detection, multidimensional fluorescence imaging, surface plasmon resonance
imaging, molecular contrast optical coherence tomography, multiscale photoacoustics, polarized light
for medical diagnostics, quantitative diffuse reflectance imaging, interferometric light scattering, non-
linear interferometric vibrational imaging, nanoscintillator-based therapy, SERS molecular sentinel
nanoprobes, and plasmonic coupling interference nanoprobes.
The three-volume handbook includes 71 chapters grouped in 8 sections:
1. Volume I: Biomedical Photonics Handbook, Second Edition: Fundamentals, Devices, and Techniques
2. Volume II: Biomedical Photonics Handbook, Second Edition: Biomedical Diagnostics
3. Volume III: Biomedical Photonics Handbook, Second Edition: Therapeutics and Advanced Biophotonics
In Volume I, Section I (Photonics and Tissue Optics) contains introductory chapters on the funda-
mental optical properties of tissue, light–tissue interactions, and theoretical models for optical imaging.
Section II (Basic Photonic Instrumentation and Use) deals with basic instrumentation and hardware
systems and contains chapters on lasers and excitation sources, basic optical instrumentation, optical
fibers, probe designs, laser use, and optical radiation safety. Section III (Photonic Detection and Imaging
Techniques) deals with methodologies and contains chapters on various detection techniques and sys-
tems (such as lifetime imaging, microscopy, two-photon detection, photothermal detection, interferom-
etry, Doppler imaging, light scattering, and thermal imaging). Finally, Section IV (Spectroscopic Data)
Preface xi

provides a comprehensive compilation of useful information on spectroscopic data of biologically and

medically relevant species for over 1000 compounds and systems.
In Volume II, Section I (Biomedical Analysis, Sensing and Imaging) contains chapters describing
in vitro diagnostics (e.g., glucose diagnostics, in vitro instrumentation, biosensors, surface plasmon
resonance, and flow cytometry) and in vivo diagnostics (optical coherence tomography, polarized light
diagnostics, functional imaging and photon migration spectroscopy, and multiscale photoacoustics).
Section II (Biomedical Diagnostics and Optical Biopsy) is mainly devoted to novel optical techniques
for cancer diagnostics, often referred to as optical biopsy (such as fluorescence, scattering, reflectance,
interferometric light scattering, optoacoustics, and ultrasonically modulated optical imaging).
In Volume III, Section I (Therapeutic and Interventional Techniques) covers photodynamic therapy
as well as various laser-based treatment techniques that are applied to different organs and disease end-
points (dermatology, pulmonology, neurosurgery, ophthalmology, otolaryngology, gastroenterology,
and dentistry). There are several chapters dealing with nanotechnology for theranostics, that is, the
modality combining diagnostics and therapy. Section II (Advanced Biophotonics and Nanophotonics)
is devoted to the most recent advances in methods and instrumentation for biomedical and biotech-
nology applications. This section contains chapters on emerging photonic technologies (e.g., biochips,
nanosensors, quantum dots, molecular probes, molecular beacons, molecular sentinels, plasmonic cou-
pling nanoprobes, bioluminescent reporters, optical tweezers) that are being developed for gene expres-
sion research, gene diagnostics, protein profiling, and molecular biology investigations as well as for
early diagnostics of disease biomarkers for the new medicine.
The goal of the second edition of this handbook is to provide a comprehensive forum that integrates
interdisciplinary research and development of interest to scientists, engineers, manufacturers, teachers,
students, and clinical providers. Each chapter provides introductory material with an overview of the
topic of interest as well as a collection of published data with an extensive list of references for further
details. The handbook is designed to present the most recent advances in instrumentation and meth-
ods as well as clinical applications in important areas of biomedical photonics. Because light is rapidly
becoming an important diagnostic tool and a powerful weapon in the armory of the modern physician,
it is our hope that this handbook will stimulate a greater appreciation of the usefulness, efficiency, and
potential of photonics in medicine.

Tuan Vo-Dinh
Duke University
Durham, North Carolina
 Acknowledgments

The completion of this work has been made possible with the assistance of many friends and colleagues.
I wish to express my gratitude to members of the Scientific Advisory Board of the first edition. Their
thoughtful suggestions and useful advice in the planning phase of the first edition have been important
in achieving the breadth and depth of this handbook. It is a great pleasure for me to acknowledge, with
deep gratitude, the contribution of over 150 contributors for the 71 chapters in this handbook. I wish to
thank my coworkers at Duke University and the Oak Ridge National Laboratory, and many colleagues
in academia, federal laboratories, and industry, for their kind help in reading and commenting on vari-
ous chapters of the manuscript. My gratitude is extended to all my present and past students, postdoc-
toral associates, colleagues, and collaborators, who have been traveling with me on this exciting journey
of discovery with the ultimate vision of bringing research at the intersection of photonics and medicine
to the service of society.
I gratefully acknowledge the support of the US Department of Energy Office of Biological
and Environmental Research, the National Institutes of Health, the Defense Advanced Research
Projects Agency, the Department of the Army, the Army Medical Research and Materiel Command,
the Department of Justice, the Federal Bureau of Investigation, the Office of Naval Research, the
Environmental Protection Agency, the Fitzpatrick Foundation, the R. Eugene and Susie E. Goodson
Endowment Fund, and the Wallace Coulter Foundation.
The completion of this work has been made possible with the love, encouragement, and inspiration of
my wife, Kim-Chi, and my daughter, Jade.

xiii
 Editor

Tuan Vo-Dinh is R. Eugene and Susie E. Goodson Distinguished

Professor of Biomedical Engineering, professor of chemistry, and
director of the Fitzpatrick Institute for Photonics at Duke University.
A native of Vietnam and a naturalized US citizen, he completed high
school education in Saigon (now Ho Chi Minh City). He continued
his studies in Europe, where he received his BS in physics in 1970
from EPFL (Ecole Polytechnique Federal de Lausanne) in Lausanne
and his PhD in physical chemistry in 1975 from ETH (Swiss Federal
Institute of Technology) in Zurich, Switzerland. Before joining
Duke University in 2006, Dr. Vo-Dinh was director of the Center
for Advanced Biomedical Photonics, group leader of Advanced
Biomedical Science and Technology Group, and a corporate fellow, one of the highest honors for distin-
guished scientists at Oak Ridge National Laboratory (ORNL). His research has focused on the develop-
ment of advanced technologies for the protection of the environment and the improvement of human
health. His research activities involve biophotonics, plasmonics, nanobiotechnology, laser spectroscopy,
molecular imaging, medical theranostics, cancer detection, nanosensors, chemical sensors, biosensors,
and biochips.
Dr. Vo-Dinh has authored over 350 publications in peer-reviewed scientific journals. He is the author
of a textbook on spectroscopy and the editor of six books. He holds over 37 US and international pat-
ents, 5 of which have been licensed to private companies for commercial development. Dr. Vo-Dinh has
presented over 200 invited lectures at international meetings in universities and research institutions.
He has chaired over 30 international conferences in his field of research and served on various national
and international scientific committees. He also serves the scientific community through his participa-
tion in a wide range of governmental and industrial boards and advisory committees.
Dr. Vo-Dinh has received seven R&D 100 Awards for Most Technologically Significant Advance
in Research and Development for his pioneering research and inventions of innovative technologies.
He has received the Gold Medal Award, Society for Applied Spectroscopy (1988); the Languedoc–
Roussillon Award, France (1989); the Scientist of the Year Award, ORNL (1992); the Thomas Jefferson
Award, Martin Marietta Corporation (1992); two Awards for Excellence in Technology Transfer, Federal
Laboratory Consortium (1995, 1986); the Inventor of the Year Award, Tennessee Inventors Association
(1996); the Lockheed Martin Technology Commercialization Award (1998); the Distinguished Inventors
Award, UT-Battelle (2003); and the Distinguished Scientist of the Year Award, ORNL (2003). In 1997,
he was presented the Exceptional Services Award for distinguished contribution to a healthy citizenry
from the US Department of Energy. In 2011, he received the Award for Spectrochemical Analysis from
the American Chemical Society (ACS) Division of Analytical Chemistry.

xv