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SEPTEMBER 2012
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Multimodal Label-Free Imaging
Drives Biomedical Research
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8 BIOSCAN
BioPhotonics editors curate the most significant headlines
of the month for photonics in the life sciences and take
you deeper inside the news. Featured stories include:
Scalpel-free surgery with deep-tissue imaging
Moth eyes inspire improved x-ray imaging
Metamolecules switch handedness under light
17 BUSINESSSCAN
Ophthalmic laser market growing
NEWS
28
BioPhotonics September 2012
20 MULTIMODAL LABEL-FREE IMAGING DRIVES BIOMEDICAL RESEARCH
by James Lopez and Yiwei Jia, Olympus America Inc.
Various optical techniques used in tandem can monitor disease processes
in real time without the shortcomings of fluorescence imaging.
24 THE AGE OF THE TRICORDER
by Gary Boas, Contributing Editor
This Star Trek-inspired medical diagnostic device is now a reality, and the
international Qualcomm Tricorder X PRIZE should help raise the bar even higher.
28 NEW GLASS FIBERS WIDEN RANGE OF MEDICAL LIGHTING APPLICATIONS
by Karen Holst, Schott AG
High-purity, chemically stable glass optical fibers offer increased
light transmission and the potential for new applications.
31 LIGHT SOURCE HELPS ENDOSCOPES GET SMALLER AND SMALLER
by James Hermanowski, Nathaniel Group Inc.
The weak delivery of xenon light through the small fibers used in
microendoscopes has driven the search for a new source.
FEATURES
www.photonics.com
Volume 19 Issue 7
6 EDITORIAL
35 BREAKTHROUGHPRODUCTS
40 APPOINTMENTS
Upcoming Courses and Shows
41 ADVERTISER INDEX
42 POST SCRIPTS
by Caren B. Les
Bioluminescence imaging lights up hair renewal
DEPARTMENTS
PHOTONICS
The technology of generating and harnessing light and other forms of radiant energy whose
quantum unit is the photon. The range of applications of photonics extends from energy generation
to detection to communications and information processing.
BIOPHOTONICS
The application of photonic products and techniques to solve problems for researchers,
product developers, clinical users, physicians and others in the fields of medicine,
biology and biotechnology.
THE COVER
While intrinsic fluorescence distinguishes individual
microvilli cells (green), a coherence anti-Stokes Raman
scattering signal (magenta) detects lipids within them
and nearby. An article on label-free imaging by James
Lopez and Yiwei Jia of Olympus America Inc. begins on
page 20. Design by Art Director Suzanne L. Schmidt.
Contents_Layout 1 8/30/12 2:26 PM Page 4
5 BioPhotonics September 2012
www.photonics.com
Group Publisher Karen A. Newman
Editorial Staff
Managing Editor Laura S. Marshall
Senior Editor Melinda A. Rose
News Editors Gary Boas, Caren B. Les, Ashley N. Paddock
Contributing Editors Hank Hogan, Marie Freebody
Copy Editors Judith E. Storie, Patricia A. Vincent,
Margaret W. Bushee
Creative Staff
Senior Art Director Lisa N. Comstock
BioPhotonics Art Director Suzanne L. Schmidt
Designer Janice R. Tynan
Director of Publishing Operations Kathleen A. Alibozek
Electronic Media Staff
Director Charley Rose
Multimedia Services & Marketing
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Web Developers Alan W. Shepherd
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Masthead_Layout 1 8/30/12 9:58 AM Page 5
Biophotonics has plenty of room
for pioneers
O
ur world has lost a number of groundbreakers in recent
days, and while it will be a different place without them,
it already is a better place because of them. Neil Arm-
strong took the first walk on the moon, and Sally Ride was the
first American woman to make the trip into space. In photonics,
Elias Snitzer is considered the father of the glass laser, and Isaac
Kaplan found a purpose for the CO
2
laser.
Kaplan, who died in August at the age of 93, was a pioneer in
the creation of the first carbon dioxide laser for general surgery
and founder of the International Society for Laser Surgery and
Medicine.
While Armstrong said that circumstance gave him the role of
first man on the moon, Kaplan had a plan. In her news report on
his death for Photonics.com, senior editor Melinda Rose writes,
When it was developed in the 1960s, Kaplan said, the CO
2
laser
was without application. His goal was to research the application
of the new miracle technology while developing an apparatus
that could put the application to work in general surgery.
(http://www.photonics.com/Article.aspx?AID=51741)
Armstrong, Ride, Snitzer and Kaplan are rarely spoken of in
the same breath, for sure, but each broke new ground in his or her
area of expertise. And, while the job of pioneer is often a one-
person enterprise, each breakthrough creates new opportunities
for exploration and innovation.
Today, biophotonics innovators receive their inspiration from
many disciplines and from all kinds of places, including yester-
days view of tomorrow, Star Trek. We have so not heard the last
of the tricorder, and that could be a very good thing for all of us.
In an article in this issue, contributing editor Gary Boas says,
The Star Trek television series and movies have proved remark-
ably prescient in anticipating the future. In The Age of the Tri-
corder, beginning on page 24, Boas describes a new generation
of sensing and imaging diagnostic devices, many with origins in
smartphones with cameras and data-transmission capabilities, and
tells us to expect to see more such devices in the future.
The brilliant sparks of inspiration that mark the early careers of
so many trailblazers are game-changing, to be sure, but its the
work that continues outside of the spotlight, often for decades,
that leads to true greatness.
Neil Armstrong also said, I guess we all like to be recognized
not for one piece of fireworks, but for the ledger of our daily
work. After his walk on the moon, Armstrong worked for NASA
and taught aerospace engineering at the University of Cincinnati.
After NASA, Sally Ride taught at the University of California,
San Diego, and encouraged girls and young women to study math
and science. Snitzer and Kaplan had long careers that included
research, innovation and education. All proved that there are
many ways to affect our world and our future.
Also in this issue, James Hermanowski of Nathaniel Group
Inc. explains that xenon light sources represent the benchmark for
medical illumination but cannot couple light through the small
channels used for microendoscopes. His article, Light Source
Helps Endoscopes Get Smaller and Smaller, starts on page 31.
In Multimodal Label-Free Imaging Drives Biomedical Re-
search, James Lopez and Yiwei Jia, of Olympus America Inc.,
describe how label-free noninvasive, nondestructive optical
microscopy using multiple simultaneous techniques allows
researchers to observe diverse life processes in real time. Find
the article on page 20.
And, finally, optical fibers with higher light transmission and
longer life spans offer interesting new solutions for meeting the
growing demand for higher quality lighting in medicine, accord-
ing to Karen Holst of Schott AG in her article, New Glass Fibers
Widen Range of Medical Lighting Applications, beginning on
page 28.
Enjoy the issue.
6 BioPhotonics September 2012
EDITORIAL
Karen A. Newman
karen.newman@photonics.com
Editorial_Layout 1 8/31/12 10:33 AM Page 6
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BioPhotonics September 2012
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from around the world.
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BIOSCAN
8 BioPhotonics September 2012
A closer look at the most significant biophotonics research and technology headlines of the month
Scalpel-free surgery with deep-tissue imaging
PASADENA, Calif. A new procedure
that more than doubles the depth that light
can be focused inside biological tissues
soon could enable doctors to perform inci-
sion-free surgery or diagnose cancer by
seeing tumors inside the body.
Although the previous limit for how
deep light could be focused into tissue was
only about 1 mm, researchers at California
Institute of Technology (Caltech) now can
reach 2.5 mm. In principle, their method
could focus light as much as a few inches
into tissue.
The ability to focus high-intensity light
tightly deep within tissue has a lot of ap-
plications, said Changhuei Yang, a pro-
fessor of electrical engineering and bio-
engineering at Caltech. We hope that
with further technology improvement,
depth up to a few centimeters will be
achievable. If we can reach up to about
10 cm, it would allow us to reach most
regions of the human anatomy.
The new technique builds on a previous
method that Yang and his colleagues de-
veloped to see through a layer of biologi-
cal tissue, which is opaque because it scat-
ters light. In that study, the scientists
shone light through a tissue sample and
recorded the resulting scattered light on a
holographic plate. The recording contained
information about how the light beam
scattered, zigzagging through the tissue.
By playing the recording in reverse, they
sent the light back through the other side
of the tissue, retracing the beams path to
the original source.
In this way, they could send light through
a layer of tissue without the blurring effect
of scattering. However, to make images of
the tissues insides, they would have to be
able to focus a beam of light into it.
To precisely focus light into tissue, the
team expanded upon the recent work of
Lihong Wangs group at Washington Uni-
versity in St. Louis (WUSTL), which de-
veloped a technique to focus light using
the high-frequency vibrations of ultra-
sound and two of ultrasounds favorable
properties. First, its high-frequency sound
waves are not scattered by tissue; second,
its ultrasonic vibrations interact with light
in such a way that the lights frequency is
shifted ever so slightly. As a result of this
acousto-optic effect, light that interacts
with ultrasound changes into a slightly dif-
ferent color.
Both teams focused ultrasound waves
into a small region inside a tissue sample
during their experiments. Next, they shone
light into the sample, which scattered the
light. Any light that passed through the re-
gion with the focused ultrasound changed
color somewhat. The researchers identified
and recorded the color-shifted light.
Using Caltechs playback technique,
they sent the light back, inducing only the
color-shifted portion to retrace the path to
the small region where the ultrasound was
focused. This means that the light itself is
focused on that area, allowing an image to
be created. By moving the ultrasounds
focus, the researchers can control where
they want to focus the light.
Only a very small amount of light could
be focused in the WUSTL experiment, but
Caltechs method allows scientists to fire a
beam of light with as much power as they
need for potential applications.
This technology is still in its infancy,
Yang told BioPhotonics. We took an im-
portant step beyond Lihong Wangs origi-
nal demonstration of TRUE (time-reversed
ultrasonically encoded optical focusing)
by implementing a TRUE technique that is
effectively unlimited in terms of its ability
to deliver arbitrarily high power to the fo-
cused spot.
For this to work in living tissue, Yang
said, the team must decrease the time for
generating a focused light spot to a frac-
tion of a second, depending on the tissue
type. The ability to build a suitable sys-
tem is within our technological reach. But
it does require a significant financial in-
vestment to make it happen. If we have
the financial resources and a semiconduc-
tor foundry to help us, bringing the tech-
nology to the point of clinical imaging is
like a 10- to 20-year process.
The team demonstrated how the new
method could be used with fluorescence
imaging by embedding a patch of gel with
a fluorescent pattern that spelled out
CIT inside a tissue sample. The investi-
A new technique more than doubles the depth that light can be focused inside biological tissue. In the
experiment, Caltech researchers shone green laser light into the tissue sample seen here in the center.
Images courtesy of Caltech/Benjamin Judkewitz and Ying Min Wang.
BioScan_Layout 1 8/30/12 12:41 PM Page 8
BioPhotonics September 2012 9
gators scanned the sample with focused
light beams, which hit and excited the
fluorescent pattern, resulting in the
glowing letters emanating from inside
the tissue. They also used the technique
to take images of tumors tagged with
fluorescent dyes.
The fluorescence is used only during
the final imaging scan, Yang said, and the
technique works whether the dye is used
or not.
We can form a focused light spot [re-
gardless], he said. We chose to do fluo-
rescence imaging here because our tech-
nique is able to generate a sufficiently
strong focused spot to excite fluorophores
to provide image contrast.
Doctors also could use the technique to
treat cancer with photodynamic therapy,
which currently can be used only at the
surface of tissue because of the way light
is easily scattered. The new method should
make it possible to reach cancer cells
deeper inside tissue.
Next, Yang said he and his team would
like to combine the spatial light modulator
and image sensor into a single digital opti-
cal phase conjugation chip, but funding is
a challenge. If they could achieve this
single chip, Yang said, it would solve
several technical implementation chal-
lenges in one move and, more importantly,
allow us to deploy sensing and playback
over a far larger area. The more area we
can cover, the task of collecting and play-
ing back the conjugate light field becomes
faster and easier.
The study appeared in Nature Commu-
nications (doi:10.1038/ncomms1925).
Left, light enters the tissue sample and is scattered
(blue arrows). From above, ultrasound is focused
into a small area inside the tissue. The ultrasound
shifts the frequency of any light that passes through
that area ever so slightly, changing its color. The
color-shifted light (green) is then recorded. Right, the
recorded light is sent back to retrace its steps to the
small region where the ultrasound was focused
which means the light itself is focused on that area.
Moth eyes inspire improved x-ray imaging
NEW YORK A new class of nanoscale materials modeled after
a moths eye could improve the light-capturing efficiency of
x-ray machines and similar medical imaging devices.
As with butterflies, moths have large compound eyes com-
posed of many thousand ommatidia structures comprising a
primitive cornea and lens, connected by photoreceptor cells.
Unlike those of butterflies, however, moth eyes are extraordinar-
ily antireflective, bouncing back only a small portion of the light
that strikes them. This adaptation makes the insects less visible
to predators during their nocturnal flights. Because of this, engi-
neers have looked to the moth eye to help design more efficient
coatings for solar panels and antireflective surfaces for military
devices.
City University of New York professor Yasha Yi and col-
leagues at Tongji University in Shanghai have taken this feature
a step further: They have used the moth eye as a model for devel-
oping nanoscale materials that someday could reduce the x-ray
radiation dosages received by patients, while improving the
resolution of the resulting images.
The scientists focused their experiment on scintillation
materials compounds that, when struck by incoming particles,
absorb the energy in the form of light. Such scintillators are used
in radiographic imaging devices to convert the x-rays exiting the
body into visible light signals picked up by a detector to form
an image.
A higher x-ray dosage improves output but is not healthy for
patients. As an alternative, Yis team found that improving the
scintillators efficiency at converting x-rays to light improved
the output. Their new nanomaterial does just that.
The material consists of a 500-nm-thick thin film composed
of a cerium-doped lutetium oxyorthosilicate crystal.
We need a thin film to fabricate the light-extraction structure,
Yi told BioPhotonics. The layer was needed so as not to perturb
the scintillation materials light emission layer.
The crystals are encrusted with tiny pyramid-shaped silicon
nitride protuberances. Each protuberance, or corneal nipple, is
modeled after the structure in a moths eye and is designed to
extract more light from the film.
Within a 100 100-m square, about the same density as
the actual moth eye, the scientists can fit between 100,000 and
200,000 protuberances. They made the side walls of the device
A scanning electron microscope image of a leaf miner moths eye. Moths large
compound eyes are extraordinarily antireflective, bouncing back only a small
portion of light that strikes them; now, researchers have used the moth eye as a
model for new nanoscale materials for improved x-ray imaging. Courtesy of
Dartmouth College.
BioScan_Layout 1 8/30/12 12:41 PM Page 9
rougher, improving its ability to scatter light and enhancing the
scintillators efficiency.
The light-extraction efficiency enhancement is very sensitive
to the dimension of the protuberances, Yi said. More research is
needed to see whether there are benefits to adding more, he said.
During lab experiments, Yi discovered that adding the thin film
to the scintillator of an x-ray mammographic unit increased the
intensity of the emitted light by as much as 175 percent compared
with the output of a traditional scintillator.
The work appeared in Optics Letters (http://dx.doi.org/10.1364/
OL.37.002808) and represents a proof-of-concept evaluation of
the use of the moth eye-based nanostructures in medical imaging
materials. It also could be applied to various light-emitting
devices, Yi said.
He estimates that it will take at least another three to five years
to evaluate and perfect the film, and five years before it will
reach a clinical setting.
The team plans to continue investigations to understand and
improve the light-enhancement mechanism.
10 BioPhotonics September 2012
b
BIOSCAN
BERKELEY, Calif. A new technique
that uses light to change the handedness
of artificial molecules could benefit tera-
hertz technology applications from bio-
medical research to ultrahigh-speed com-
munications and homeland security.
Using a light beam, the chirality of arti-
ficial molecules has been switched from a
right-handed orientation to a left-handed
one for the first time. Chirality is the dis-
tinct left or right orientation, or handed-
ness, of some types of molecules mean-
ing it can take one of two mirror-image
forms. Called enantiomers, the right- and
left-handed forms of such molecules can
exhibit strikingly different properties; for
example, one enantiomer of the chiral
molecule limonene has a lemon scent,
while the other smells of orange.
The ability to observe or switch a mole-
cules chirality using terahertz electromag-
netic radiation is a coveted asset in high
technology.
In electromagnetism, chirality or opti-
cal activity arises from the coupling be-
tween the electric and magnetic responses
of the materials, said Xiang Zhang, one
of the leaders of the research and a princi-
pal investigator with the US Department
of Energys Lawrence Berkeley National
Laboratorys Materials Sciences Div.
However, in natural materials, the mag-
netic response is extremely weak at THz
and optical frequencies, and as a result,
the chirality is also very weak.
Using terahertz metamaterials engi-
neered from nanometer-size gold strips
with air as the dielectric, Zhang and a
multi-institutional team of colleagues from
Los Alamos National Lab and the Univer-
sity of Birmingham in the UK fashioned a
delicate artificial chiral molecule that they
incorporated with a photoactive silicon
medium. By performing photoexcitation of
their metamolecules with an external light
beam, they observed dynamically con-
trolled handedness flipping in the form of
circularly polarized emitted terahertz light.
Under strong optical irradiation, the
handedness of the metamolecule is
switched to its opposite handedness,
Zhang told BioPhotonics. This state is
temporary; it relaxes back to its original
handedness in a time scale of 1 millisec-
ond. The process is repeatable.
The optically switchable chiral terahertz
metamolecules consisted of a pair of 3-D
meta-atoms of opposite chirality made
from precisely structured gold strips. Each
meta-atom serves as a resonator with a
coupling between electric and magnetic
responses that produces strong chirality
and large circular dichroism at the reso-
nance frequency.
When two chiral meta-atoms of the
same shape but opposite chirality are put
together, they form a metamolecule, and
their symmetry is preserved, resulting in
vanishing optical activity. Essentially,
the optical activity that arises from the
opposite meta-atoms cancels each other
out, he said.
To break the mirror symmetry and in-
duce chirality for the combined metamole-
cule, the researchers introduced silicon
pads to each chiral meta-atom in the meta-
molecule. In one meta-atom, the silicon
pad bridged two gold strips, while the sili-
con pad replaced part of the gold strip in
the other meta-atom. The silicon pads
functioned as the optoelectronic switches
that flipped the chirality of the metamole-
cule under photoexcitation.
Terahertz electromagnetic radiation falls
within the frequency range of molecular
vibrations, making it a suitable noninva-
sive tool for analyzing the chemical con-
stituents of organic and nonorganic mate-
rials. By having the ability to flip the
handedness of metamolecules and control
the circular polarization of terahertz light,
scientists could use the technology to de-
Metamolecules switch handedness under light
Controlling the chirality of artificial molecules could
enable advances in communications and biomedical
imaging. Top, a scanning electron microscope image
of optically switchable chiral terahertz metamole-
cules. Bottom, the purple, blue and tan colors
represent the gold meta-atom structures at different
layers; two silicon pads are shown in green. Images
courtesy of Xiang Zhang et al, Berkeley Lab.
BioScan_Layout 1 8/30/12 12:41 PM Page 10
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BIOSCAN
In this schematic, the chirality-switching metamolecule consists of four chiral resonators with fourfold
rotational symmetry. An external beam of light instantly reverses the metamolecules chirality from
right-handed to left-handed.
tect toxic or explosive chemicals, or use it
in high-speed data processing systems and
wireless communications.
Terahertz-based polarimetric devices
also could benefit medical researchers and
developers of pharmaceutical drugs be-
cause most biological molecules, includ-
ing DNA, RNA and proteins, are chiral.
In THz, most of the biological mole-
cules show circular dichroism, Zhang said.
However, there is a lack of spectroscopy
tools to accurately measure the circular
dichroism of biomolecules at THz in com-
parison with the visible range. The sensitive
detection of circular dichroism requires dy-
namic modulation of the electromagnetic
waves between the two circular polariza-
tions. The chirality-switching metamaterial
we demonstrated may bridge this gap.
Their design principle for optically
switchable chiral terahertz metamolecules
is not limited to just handedness switching;
it also could be applied to dynamic revers-
ing of other electromagnetic properties.
Dynamically reversing other electro-
magnetic properties would enable us com-
plete control of electromagnetic waves, not
only in polarization, but also in phase, in-
tensity and propagation directions, Zhang
said. For example, we can use a similar
design principle to make a meta-surface
with dynamically switchable high and low
impedance. For a THz wave reflected by
the meta-surface, the phase can be dynami-
cally switched between 0 and 180 degrees.
The metamaterials we demonstrated so
far still have some drawbacks: Chirality is
not strong enough to completely convert
the THz waves into purely circular polar-
izations. We will work on the perfection of
the metamaterial design to achieve a
stronger chirality switching effect.
The work was published in Nature Com-
munications (doi:10.1038/ncomms 1908).
BioScan_Layout 1 8/30/12 12:41 PM Page 11
LOS ANGELES A new optical micro-
scope easily detects rare cells in real time
and could help doctors diagnose diseases
earlier or monitor treatments. The ability
to distinguish and isolate rare cells such
as circulating cancer tumor cells and stem
cells from a large population of assorted
cells has become increasingly important
for disease detection. Typically, there are
only a handful of these rogue cells
among a billion healthy ones, but because
they are precursors to metastasis the
spread of cancer that causes about 90 per-
cent of cancer mortalities it is important
to be able to find them.
To detect such cells requires an auto-
mated high-throughput instrument that can
examine millions of cells in a reasonably
short time. Currently, microscopes
equipped with digital cameras are used to
analyze cells, but they are too slow to be
useful for this application.
Now, Bahram Jalali and Dino Di Carlo
from the University of California, Los An-
geles, have devised a high-throughput
flow-through optical microscope that can
detect rare cells with sensitivity of one
part per million in real time. The instru-
ment is equipped with photonic time-
stretch camera technology developed by
Jalalis group in 2009 to create the quick-
est continuous-running camera in the
world.
To catch these elusive cells, the camera
must be able to capture and digitally pro-
cess millions of images continuously
at a very high frame rate, said Jalali, who
holds the Northrop Grumman Endowed
Opto-Electronic Chair in Electrical Engi-
neering at the UCLA Henry Samueli
School of Engineering and Applied Sci-
ence. Conventional CCD and CMOS
cameras are not fast and sensitive enough.
It takes time to read the data from the
array of pixels, and they become less sen-
sitive to light at high speed.
The team described how it integrated
the camera with real-time image process-
ing and advanced microfluidics for the
classification of cells in blood samples in
Proceedings of the National Academy of
Sciences (doi 10.1073/pnas.1204718109).
The new blood-screening technology de-
livers throughput of 100,000 cells per sec-
ond, roughly a hundredfold increase in
rate when compared with conventional im-
aging-based blood analyzers.
This study illustrates the detection of
rare breast cancer cells in blood in real
time with an unprecedented low false-pos-
itive rate of one cell in a million. Initial re-
sults show that this method can quickly
detect rare circulating tumor cells from
large volumes of blood samples, opening
the door for statistically accurate early
cancer detection and for monitoring the ef-
ficacy of drug and radiation therapies. The
results were obtained by mixing cancer
cells grown in a laboratory with various
proportions of blood to emulate real pa-
tient blood.
The team is conducting clinical trials to
assess the efficacy of the technology,
which can significantly reduce errors and
costs in medical diagnosis. It also could be
used for water-quality monitoring and
urine analysis.
The research was funded by the US
Congressionally Directed Medical Re-
search Programs, the Burroughs Wellcome
Fund and NantWorks LLC.
12 BioPhotonics September 2012
b
BIOSCAN
For more on the camera technology de-
veloped by Bahram Jalalis group, see
Full Steam Ahead with the Fastest
Camera in the World, www.photonics.
com/a38998.
Optical microscope detects rare cancer cells
Optical microscope with worlds fastest camera. Courtesy of UCLA.
Photonic crystals help fish see in the murk
LEIPZIG, Germany The elephantnose
fish has light-reflecting cups lined with
photonic crystals in its retinas that help it
navigate its dark, murky environment, sci-
entists have found. This unusual eye struc-
ture might aid future sensors that filter sig-
nal noise or peform detection in turbid
fluids.
These freshwater fish, found in the
cloudy depths of African rivers, use elec-
trosensing to navigate their dark environ-
ment, but they also depend somewhat on
vision. Until recently, these weakly elec-
tric fish were thought to be blind, or al-
most blind, University of Leipzig neuro-
physiologist Andreas Reichenbach said in
a podcast.
The visual capabilities of this fish are
pretty poor, he said. Its color-blind. It
cannot see anything that is not bigger than
six times the size of a full moon. But sur-
prisingly, its optimal for its environment.
Vertebrate eyes have rod photorecep-
tors, which are very sensitive to light but
do not detect color or fine details, and
cone photoreceptors, which are less sensi-
tive to light but can distinguish color and
details. Most vertebrate eyes optimize pri-
marily either rods or cones, but an interna-
tional team of scientists has discovered
BioScan_Layout 1 8/30/12 12:41 PM Page 12
that the retina of the elephantnose fish is
structured so that both types of receptors
work simultaneously.
The researchers observed that the cones
are grouped together within larger, light-
reflecting cups lined with photonic crys-
tals. The rods are positioned behind these
reflectors. This unusual arrangement
works because the mirrorlike surfaces of
the cups propagate the light in a way that
delivers just the right wavelength to both
the rods and cones.
Behind the eye in the so-called retina
pigment epithelium, there are huge cells,
which form kind of parabola mirrors re-
flecting the light, Reichenbach said.
From outside, you can see that light is re-
flected, like in the cats eye, but it is fo-
cused on a certain level in this case. The
astonishing thing is that within such
parabola mirrors, there are about 30 cone
photoreceptors, which are responsible for
high-acuity vision in our case, but not in
this case, and a couple hundred rod pho-
toreceptors.
Each photoreceptor sees the same part
of an image because all the rods and cones
are illuminated by the same structure,
meaning that the visual acuity is very bad,
he said. The fish are at an advantage be-
cause they are not able to see or get dis-
tracted by the small particles, such as dead
worms and bubbles or air moving around
them, but they can see the big predators
moving.
The special structure of the retina
of the fish enables the fish to see large
moving objects more reliably than any
other fish, and this makes him thrive
under these [turbid water] conditions,
he said.
While this vision proves advantageous
for the fish in dark, dim, muddy waters,
its detrimental in aquarium settings
with other fish.
If the fish swims in an aquarium
together with other fish under normal
laboratory conditions, its almost blind,
Reichenbach said. This was the reason
why the fish had been considered blind
for many years.
The study appeared in Science (doi:
10.1126/science.336.6089.1617-c).
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BioPhotonics September 2012 13
b
BIOSCAN
Scientists had long thought the elephantnose fish (Gnathonemus petersii) to be blind, but it turns out to have
light-reflecting cups lined with photonic crystals in its retinas that help it navigate in dark waters. Courtesy of
Gerhard von der Emde.
Light amplification by photonic-crystal light collector
onto photoreceptor cells in the retina of the ele-
phantnose fish helps it to see. Courtesy of Moritz
Kreysing.
BioScan_Layout 1 8/30/12 2:28 PM Page 13
LEUVEN, Belgium A new nanoscale
light-manipulation method that optically
detects single molecules could be used in
a variety of photochemistry applications
and help advance technologies for visual-
izing single molecules and multiple-mole-
cule interactions.
Progress in optically detecting single
molecules has been hindered by their weak
optical response. Currently, researchers use
metal nanostructures to focus light into
tiny zones called hot spots, which excite
electrons on the surface, causing them to
oscillate coherently. When shone on a mol-
ecule, and with the help of these oscillating
electrons, the focused light can increase a
molecules optical signal to 100 billion
times its normal strength, a level detectable
by optical microscopes.
The current method, however, has two
limitations: The first is that hot spots can
become too hot; the second is that they are
very small. This means that the heat from
hot spots can melt the nanostructure, de-
stroying its ability to channel light effec-
tively. And hot spots produce only a very
small cross section in which interaction
with molecules can take place. For a sin-
gle molecule to become detectable, it
must find the hot spot.
To overcome these drawbacks, Dr.
Ventsislav Valev and colleagues at
Katholieke Universiteit Leuven sought to
nanoengineer larger spots. The interna-
tional team began by shining circularly
polarized light on nanostructures and
found that this could increase the useful
area. When they shone light on square-
ring-shaped gold nanostructures, the scien-
tists observed that the entire surface of the
nanostructures was successfully activated.
Essentially, light is constituted of elec-
tric and magnetic fields moving through
space, Valev said. While with linearly
polarized light the fields move in a linear,
forward direction, with circularly polar-
ized light, they rotate in a spiral-like
motion.
The circularly polarized light imparts a
sense of rotation on the electron density in
ring-shaped gold nanostructures, thus trap-
ping the light in the rings and forming
loops of light. The loops cause excited
electrons to oscillate coherently on the full
surface of the square-ringed nanostruc-
tures, rather than in a few concentrated hot
spots. This increases the opportunity for
interaction with molecules.
The trick is to try to activate the whole
surface of the nanostructure so that when-
ever a molecule attaches, we will be able
to see it, Valev said. That is precisely
what we did.
The study appeared online in Advanced
Materials (doi: 10.1002/adma.201201151).
14 BioPhotonics September 2012
b
BIOSCAN
Shining circularly polarized light on ring-shaped nanostructures increases the opportunity for interaction with
molecules. Courtesy of Katholieke Universiteit Leuven.
Loops of light optically detect single molecules
Antennas capture, upconvert weak IR light
GRONINGEN, Netherlands A new tech-
nique that uses special molecules as light
antennas to harvest the energy from weak
infrared light and amplify the process
3300 times could lead to improved med-
ical imaging methods.
Materials scientists and chemists from
the University of Groningen and from the
FOM Foundation harvested infrared light
which has too little energy to release
electrons in solar cells more efficiently
by modifying an organic dye that acts as
light antennas to transmit the energy to the
nanoparticles to which they are attached.
These particles subsequently convert
two weak captured photons into a
single strong, energy-rich photon in a
Inspiration from nature: (left) a natural photosynthesis system with light-harvesting (LH) molecules and a reac-
tive center (RC); (right) a schematic representation of the nanocrystal that realizes the upconversion (UC) with
the attached antennas in green. Courtesy of University of Groningen.
BioScan_Layout 1 8/30/12 12:42 PM Page 14
process called upconversion.
There are inorganic materials made
from rare-earth metals that can facilitate
this upconversion process, said Jan C.
Kees Hummelen, a University of
Groningen professor of organic chemistry
and leader of the FOM focus group on
next-generation organic photovoltaics.
However, these materials absorb very
few infrared photons. We have therefore
attached organic molecules to them [as an-
tennas] that can capture these photons and
transmit the energy to the upconversion
material.
Because of this, the entire infrared ab-
sorption process, upconversion and the
emission of visible light is increased by a
factor of 3300, Hummelen said.
Even with the antennas, his group can
capture only a limited amount of infrared
light. He predicts that an even better yield
can be obtained, but because the upcon-
version process inside the nanocrystal is
still inefficient, it is not yet possible to
achieve.
Two photons must come together in
the material within a short space of time,
he said. In practice, the efficiency of this
process is still very low. However the har-
vest is already much better, so step one
has been achieved.
The upconversion system could be ap-
plicable for medical imaging techniques.
Infrared light penetrates further into
biological tissues than visible light, he
said. If you allow compounds that carry
out upconversion to bind to specific cells
in tissues, then you can make images
using infrared light.
The scientists work also is applicable
to solar cells, as about half of all the solar
energy reaching Earths surface consists of
infrared light.
The research was published online in
Nature Photonics (doi: 10.1038/npho
ton.2012.158).
BioPhotonics September 2012 15
b
BIOSCAN
Nanohole-based sensors ideal for medical diagnostics
SINGAPORE Novel molecular sensors
based on thin metallic films with nano -
holes hold promise for applications that
require detection of small quantities of
molecules, such as gas biomedical diag-
nostics and gas sensing.
The majority of these applications call
for inexpensive disposable sensors, but
they must be sensitive enough to detect
single molecules.
Ping Bai and colleagues at A*Star Insti-
tute of High Performance Computing and
the Institute of Materials Research and En-
gineering have studied the properties of
thin-film metallic films with holes in them
that show promise for molecular sensing
applications.
Metallic thin films with nanometer-size
holes are known to transmit light of partic-
ular wavelengths efficiently because of
surface plasmon polaritons (SPPs), the
BioScan_Layout 1 8/30/12 12:42 PM Page 15
collective movement of electrons on a
metal surface that focus light into tiny
spots much smaller than the wavelength of
light used.
SPPs can detect molecules through the
fluorescence of tracer molecules attached
to them. The SPPs enhance the fluores-
cence, which is easily detectable by a mi-
croscope, even for small quantities of mol-
ecules.
The whole setup is ultracompact to
support a point-of-care sensing system,
Bai said.
The team studied two sensing arrange-
ments: In the first, light was directed at a
film with nanoholes at an oblique angle
from the same side as the sample. The
film in the second arrangement was illu-
minated from the back so that light trav-
eled through the holes first. The re-
searchers observed that both arrangements
had advantages.
In the reflection scheme, the SPP ef-
fect is stronger because the light is aimed
directly at the sample and does not have to
cross the metal film. However, a thicker
film is needed so that the light does not
pass through. The intensity of the light
emitted by the molecules is weaker in the
transmission scheme, but filters and
other sensors could be included with the
metal film, and the film could be much
thinner.
There is therefore no clear advantage
for either sensing mode of such films,
Bai said. One thing that is clear from the
study, however, is the clear benefits of
using metal films with nanoholes as a
molecular sensing platform.
This is only a snapshot of the entire
project, Bai said.
Ultimately, our sensing technology
will be utilized in hospitals and test cen-
ters; for example, in prostate cancer
screening, or even used at home just like
glucose test kits.
The research appeared in IEEE Photon-
ics Journal (doi: 10.1109/jphot.2011.217
7652).
16 BioPhotonics September 2012
b
BIOSCAN
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In surface plasmon polariton (SPP) sensing, nanohole films can be used in two configurations to sense
molecules in a water solution. In the reflection mode (top), light is directed at the sample from the water side.
In the transmission mode (bottom), it is directed at the sample from the back, leading to different SPP proper-
ties. The SPP field intensity is represented by the color plot; the optical fields on the top and bottom are
calculated for different resonance frequencies. 2012 IEEE.
Ashley N. Paddock
ashley.paddock@photonics.com
BioScan_Layout 1 8/30/12 12:42 PM Page 16
BUSINESSSCAN
As the population of the world ages, asso-
ciated eye problems will mean significant
growth for the ophthalmic laser market.
The number of people 60 and older has
tripled in the past 50 years. As of 2000,
there were 606 million people over the age
of 60 worldwide, according to a United
Nations report, and that global population
is projected to reach nearly 2 billion in
2050.
In the US alone, the 90-and-older popu-
lation nearly tripled over the past three
decades, reaching 1.9 million in 2010, ac-
cording to a November 2011 report by the
US Census Bureau and supported by the
National Institute on Aging. Over the next
four decades, this population is projected
to more than quadruple.
The aging population is subject to
unique eye conditions, such as cataracts,
which are a clouding of the eyes natural
lens. More than 15 million cataract surger-
ies are performed worldwide each year to
surgically replace the damaged lens with
an intraocular lens.
Besides the aging population, other fac-
tors driving the global ophthalmic laser
market, which is expected to reach $804
million by 2015, are increasing accessibil-
ity to advanced laser eye treatment; the in-
creasing proportion of people needing vi-
sion correction, especially in Asia; and an
increase in patients opting for eye surgery,
said Global Industry Analysts (GIA) Inc.
in an ophthalmic lasers report released in
January.
Although demand for certain major
ophthalmic laser treatments declined dur-
ing the economic recession as people post-
poned elective surgeries, technological de-
velopments continued, GIA said,
providing the stimulus needed to drive fu-
ture growth.
Certain new technological develop-
ments in laser surgery are believed to offer
benefits beyond vision correction and at-
tract significant demand in the future,
GIA said. Application of laser technology
in early diagnosis, and the detection of
certain eye disorders in conjunction with
imaging technologies, such as OCT, is an-
other factor that would boost growth of
the laser eye correction market in the fu-
ture.
On Aug. 6, New York-based eye health
company Bausch + Lomb and ophthalmic
laser maker Technolas Perfect Vision of
Munich announced that their Victus fem-
tosecond laser designed for cataract and
corneal surgery had received clearance
from the FDA.
Femtosecond lasers, with their ultra-
short pulses, do not transfer heat or shock
to the material being cut and can make
surgical incisions with extreme precision.
The technology was introduced commer-
cially in 2002 for creating thin, hinged
flaps during lasik surgery.
Companies that make femtosecond
lasers commercially for ophthalmic appli-
cations include Calmar Laser, Advanced
Medical Optics and Carl Zeiss Meditec.
Carl Zeiss Meditec announced in April
that it will begin a US clinical trial of its
ReLEx smile procedure for correcting my-
opia, or nearsightedness, after receiving
FDA approval.
In lasik procedures, the excimer laser
vaporizes tissue, but the ReLEx smile
method generates a refractive lenticule in
the intact cornea with a femtosecond laser.
The surgeon then removes the lenticule
through a small incision less than 4 mm
without having to move the patient to an
excimer laser, the company said.
The introduction of femtosecond laser
technology is the most significant ad-
vancement in cataract surgery in recent
history, said Dr. Steven J. Dell of Dell
Laser Consultants in Austin, Texas, in the
press release announcing the Victus ap-
proval.
The Victus is the first femtosecond laser
that can support both surgical procedures
on a single platform, the companies say,
and it is designed to provide greater preci-
sion compared with manual cataract sur-
gery techniques. The laser received CE
mark approval in Europe in November
2011 and has been used in more than 2000
cataract or refractive procedures world-
wide, the companies say. They are work-
ing to gain approval in the US for addi-
tional applications.
In June, Iridex Corp. announced the
first use of its MicroPulse laser therapy
(MPLT) through an intraocular fiber optic
endoprobe during ophthalmic surgery.
Expanding MPLT applications from
physicians offices into the operating room
and surgery centers will continue to drive
growth in our laser systems, said Iridex
President and CEO Dominik Beck.
MicroPulse works by electronically
chopping a continuous-wave laser emis-
sion into trains of microsecond pulses, en-
hancing the physicians ability to more
precisely control the laser effects on target
tissues. It is more effective for very thin
retinas, allowing more tissue to be pre-
served than in conventional continuous-
wave laser photocoagulation, the company
said.
Iridex is working to accelerate adoption
of MicroPulse for treating diabetic macu-
lar edema (DME), Beck said in an early
August statement announcing the com-
Ophthalmic laser market growing
BioPhotonics September 2012 17
BussinessScan_Layout 1 8/30/12 9:59 AM Page 17
panys second-quarter financial results.
Our experiences in DME have led us
to explore opportunities for MicroPulse in
the treatment of glaucoma, he said,
adding that the company is working with
key opinion leaders in glaucoma on how
its product can improve the current stan-
dard of care for treating the disease.
The new glaucoma therapy, a tissue-
sparing, repeatable technique called Mi-
croPulse Laser Trabeculoplasty, was intro-
duced by Iridex at the recent American
Society of Cataract and Refractive Surgery
meeting.
If the results of these initiatives indi-
cate an equivalent or better outcome than
currently available options, we could see a
significant impact to our revenues, as this
is a large market opportunity, Beck said.
The companys revenue from ophthal-
mology was $8.4 million for the quarter,
up from slightly over the prior quarter and
the prior year.
Dental laser maker Biolase also is see-
ing opportunity in the ophthalmic market.
We continue to seek the most efficient
route to market for our laser technology as
it relates to ophthalmology and other sur-
gical specialties, said Federico Pignatelli,
chairman and CEO, during an earnings
call on Aug. 8. We are currently in early
discussions with a leading ophthalmic
company to develop a relationship to
jointly deliver our first product to that
marketplace.
18 BioPhotonics September 2012
b
BUSINESSSCAN
Melinda A. Rose
melinda.rose@photonics.com
BUSINESSBRIEFS
ON Semiconductor Corp. of Phoenix and
Donald Colvin, its executive vice president and
chief financial officer, have mutually agreed to
Colvins resignation as an officer of the com-
pany. It was expected at the time of the an-
nouncement in August that he would continue
at the firm for up to 90 days while the board
sought a replacement. ON Semiconductor sup-
plies silicon solutions for energy-efficient elec-
tronics. Its power and signal management,
logic, discrete and custom devices help cus-
tomers solve their design challenges in automo-
tive, computing, LED lighting, medical, military,
aerospace, consumer and power applications.
Laser-sintering systems manufacturer EOS of
Krailling, Germany, filed a patent lawsuit March
5 against Phenix Systems of Riom, France, for
infringing two US patents for its dental product
lines. The lawsuit alleges infringement of US
Patent Nos. 5,753,274 and 6,042,774 through
the manufacture, sale and use of the PXL, PXM,
PXS and PXS Dental product lines from Phenix in
the US. During the second half of 2011, Phenix
announced publicly the commercial manufac-
ture, sale and use of exactly these product lines,
even though, EOS said, it had apprised Phenix
of its patent portfolio several times.
To honor professor Michael S. Felds fundamen-
tal contributions in the fields of laser science
and applied physics for solving biomedical
problems, The Optical Society (OSA) and the
OSA Foundation, both in Washington, have
established the Michael S. Feld Biophotonics
Award to be given annually beginning in 2013.
It will recognize individuals for innovative and
influential contributions to the field of biopho-
tonics, regardless of their career stage. Feld,
who passed away in 2010, founded the Laser
Biomedical Research Center at MIT in 1985
with the support of the National Institutes of
Health. Nominations for the award are now
being accepted from OSA members.
Shimadzu Scientific Instruments of Colum-
bia, Md., has opened the Shimadzu Center
for Advanced Analytical Chemistry at the
University of Texas at Arlington to give re-
searchers there access to enhanced capabilities
for trace qualitative and quantitative analysis.
The center contains $6 million worth of chro-
matography, mass spectrometry and spec-
troscopy equipment. The instruments will be
used to research illnesses such as cancer and
malaria, and to develop nanofabrication
materials for industry. Kevin Schug has been
named the Shimadzu Distinguished Professor
of Analytical Chemistry. An associate professor
of chemistry and biochemistry, he will oversee
the new laboratory.
DigitalOptics Corp. of San Jose, Calif., a
subsidiary of Tessera Technologies Inc., and
MMD Monitors and Displays Taiwan Ltd., a
subsidiary of TPV Technology Ltd., have part-
nered to deliver a computer monitor manufac-
tured by MMD with a built-in ergonomic sensor
to help users correct posture. TPV owns part of
Philips monitor and entry-level flat-screen TV
business, and DigitalOptics developed the soft-
ware used in Philips ErgoSensor detection tech-
nology. The technology uses the distance be-
tween a users pupils to determine whether he
or she is sitting within the optimal ergonomic
zone and, if not, determines how the user can
adjust seating position or posture to avoid risk
of injury. In addition, the device offers energy
savings by powering down when no user is
present.
Genia Photonics of Laval, Quebec, Canada,
has received $300,000 in financial assistance
from the Canadian government to acquire
state-of-the-art production equipment and labo-
ratory materials. The repayable funding was
awarded under the Canada Economic Devel-
opments business and regional growth pro-
gram and could result in the creation of 22 jobs
by 2014. The new equipment and laboratory fa-
cilities will enable the company to meet growing
customer demand and to accelerate research
and development work it has initiated over
the past two years. Genia Photonics develops
and manufactures pulse-programmable and
multifunctional fiber-based lasers for security,
biomedical, pharmaceutical and chemical
applications.
Three photonics companies have received the
Queens Award for Enterprise, the UKs most
coveted award for business success. Edinburgh
Instruments of Livingston, a photonics and
electro-optical scientific instrumentation manu-
facturer, received the honor for its sustained
international growth. Laser Quantum of
Cheshire was recognized for providing laser
technology equipment and services to the
aerospace, medicine, research and biomedical
sectors. Fianium of Southampton received the
award in the innovation category for its devel-
opment of the WhiteLase supercontinuum fiber
laser. The awards are made each year by the
queen on the advice of the prime minister and
an advisory committee.
Raptor Photonics Ltd. of Larne, Northern Ire-
land, has named Laser 2000 GmbH of Munich
as its scientific camera distributor for Germany
and Austria. Laser 2000 supplies lasers and
light sources, optics and optomechanics, optical
instrumentation and optical detectors for the
machine vision and scientific and instrumenta-
tion markets. Raptor Photonics manufactures
industrial-grade low-light digital and analog
cameras.
Given Imaging Ltd.s patent infringement
lawsuit against Intromedic Co. Ltd. of Seoul,
South Korea, has been upheld by the Intellec-
tual Property Tribunal of the Korean Intellectual
Property Office (KIPO). Given Imaging of
Yokneam, Israel, alleges that Intromedics cap-
sule endoscope, marketed under the brand
name MiroCam, has infringed two of its Korean
patents. In response to the accusation, In-
tromedic brought proceedings before KIPO to
invalidate the two patents asserted by Given
Imaging. KIPOs decision rejects Intromedics
invalidity arguments and cripples one of its
main defenses against Given Imagings patent
infringement action. The decisions may be
appealed by Intromedic.
Olympus America Inc. of Center Valley, Pa.,
and Carl Zeiss Microscopy GmbH of Jena,
Germany, have signed a nonexclusive world-
wide licensing agreement allowing the German
company to access Olympus portfolio of digital
pathology and virtual microscopy patents. The
patents in the licensing deal cover methods and
equipment for creating, storing and delivering
virtual microscopy slides. The technology en-
BussinessScan_Layout 1 8/30/12 9:59 AM Page 18
ables individuals to view and share high-resolu-
tion virtual microscopy images over the Internet.
Specific terms and conditions of the agreement
will not be made public. The microscopy busi-
ness group at Carl Zeiss manufactures light and
electron microscopes for the life and materials
sciences.
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Sam Mansour, who performed the surgery at
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state laser manufacturer Crystal Laser Sys-
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BioPhotonics September 2012 19
BUSINESSSCAN
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BussinessScan_Layout 1 8/30/12 9:59 AM Page 19
O
ptical microscopy is a power-
ful tool in bioscience research,
allowing researchers to ob-
serve multiple complex physi-
ological processes in real time. Fluores-
cence imaging a major technique used
with optical microscope systems pres-
ents many challenges, however. Achieving
sufficient concentration of a fluorescent
probe within a tissue can be difficult. Photo-
toxicity caused by singlet oxygen forma-
tion is damaging to living cells. And, in
live animal and animal tissue imaging, the
probe is likely to be recognized as a for-
eign entity by the animals immune
system.
Multimodal label-free imaging includ-
ing second-harmonic generation (SHG),
coherent anti-Stokes Raman scattering
(CARS) and intrinsic two-photon excita-
tion fluorescence offers a technique to
monitor disease processes over time with-
out perturbing physiological processes
by incision, adding foreign reagents or
encountering artifacts caused by fixation,
permeabilization or biochemical extraction
protocols. There is also an advantage in
using label-free imaging techniques based
on harmonics or Raman scattering because
electrons are not excited in the same man-
ner as in regular fluorescence excitation.
In both harmonics and Raman scattering
imaging, the formation of reactive oxygen
and its subsequent toxicity are signifi-
cantly reduced.
Multimodal imaging
Multimodal label-free imaging exploits
the properties of molecules when they in-
teract with photons from the short-pulsed
lasers used in multiphoton microscope
systems. These two-photon or multiphoton
imaging systems typically are optimized
for observation of fluorescent molecules
several hundred microns deep in tissue,
exploiting the improved penetration and
reduced scattering of longer infrared
wavelengths. To achieve the two-photon
effect, these lasers produce an intense flux
of photons in femtosecond pulses. The
total energy is kept low to avoid overheat-
ing the sample.
The only position where the photon flux
is strong enough for two or more photons
to combine their energies is at the exact
focal point. Because there is no excitation
outside that spot, fluorescence is confined
to molecules in one optical section as the
laser scans over the field of view. This in-
herent optical sectioning also works with
Multimodal Label-Free Imaging
Drives Biomedical Research
20 BioPhotonics September 2012
Label-free noninvasive, nondestructive optical microscopy
using multiple simultaneous techniques allows researchers
to observe diverse life processes in real time.
Figure 1. Multimodal imaging of macromolecules in C. elegans. Using multiphoton excitation at 800 nm,
a second-harmonic generation signal was collected at 375 to 405 nm in both the reflected (top left) and
forward (top right) external nondescanned detectors. Intrinsic two-photon excitation fluorescence (autofluores-
cence) was simultaneously collected at 520 to 560 nm in the reflected external nondescanned detectors with
800-nm excitation (bottom left, with overlay at bottom right). Maximum intensity projections of 139 Z-series
slices acquired at 0.4 m per optical slice. Scale bar 50 m. Courtesy of Delong Zhang, Purdue University.
BY JAMES LOPEZ AND YIWEI JIA, OLYMPUS AMERICA INC.
Feat Olympus_Layout 1 8/30/12 10:00 AM Page 20
other photonic techniques that depend on
these photon interactions, including har-
monic generation and CARS.
Multimodal imaging is the combination
of two-photon techniques such as two-
photon excitation fluorescence of intrinsi-
cally fluorescent molecules, SHG and
CARS in a single experiment. When com-
bined, these techniques create a more
complete image of life processes without
additional labels.
Most cells and tissues exhibit a complex
mixture of spectra from autofluorescence
(also called intrinsic fluorescence). It is
possible to attribute intrinsic fluorescence
of a particular wavelength to molecules
such as nicotine adenine dinucleotide
(NADH), lipofuscin or elastin. The peak
emission for these molecules is in the blue
range due to their component aromatic
amino acids, so intrinsic fluorescence is
often imaged using multiphoton excitation
rather than a blue or UV-emitting laser to
avoid the photodamage with the higher-
energy (shorter wavelength) photons.
SHG occurs when two photons are scat-
tered simultaneously by an orderly array
of molecules that all point in the same di-
rection. Known as inversion asymmetry,
this type of molecular structure is found
in various tissue structures including colla-
gen fibers, myofilaments and microtubules
of the mitotic spindle. The second-har-
monic signal is emitted coherently in
the direction of the incident beam, with
exactly twice the incoming energy and
therefore half the wavelength.
In CARS, the three photons that interact
originate with two different wavelengths.
The difference in frequency gives rise to a
signal because of resonance with specific
chemical bonds, notably the C-H bond
stretch that predominates in lipids.
Methodology
A laser-scanning microscope equipped
for multiphoton excitation can be used to
image both fluorescence and SHG through
the selection of appropriate excitation
wavelengths and filters. Intrinsic fluores-
cence is a hundredfold weaker than most
exogenous fluorophores, so the detection
path must be optimized by using high-
numerical-aperture objective lenses and
nondescanned detectors.
SHG signals typically are stronger in
the forward direction, so a transmitted
nondescanned fluorescence detector and
high-numerical-aperture condenser with an
oil-top lens is preferred. Forward-directed
SHG detection tends to favor large, bright
structures, while the reflected direction is
often more sensitive to finer, dimmer
structures. This is also generally true for
CARS imaging. Muscle fibers can be vi-
sualized without the use of any labels in
Caenorhabditis elegans, a commonly used
model organism in biological research,
using SHG. SHG and intrinsic two-photon
excitation fluorescence (autofluorescence)
signals can be collected simultaneously in
both the forward and reflected directions,
and then can be combined into a single
multimodal imaging data set (Figure 1).
There are several ways to generate the
two beams that combine to produce the
CARS signal: Researchers have often used
two picosecond-pulsed IR lasers tuned to
BioPhotonics September 2012 21
Figure 2. Green fluorescent protein expressing tumor growing in liver, imaged
using simultaneous multimodal imaging. Collagen fibers (white) are visualized
with second-harmonic generation. Lipids are imaged using femto-CARS (red).
The GFP-labeled tumor mass and the intrinsic fluorescence are co-observed in
the green channel. Maximum intensity projection of a stack of 12 Z-planes
spaced at 2.42 m. Courtesy of Teng-Leong Chew, Northwestern University.
Figure 3. Lipids exterior to and within cells after mice were gavaged with 200 l
of oleic acid. The CARS signal (magenta) from lipids and intrinsic fluorescence
(green) delineates individual cells of the microvillus. Maximum intensity projec-
tion, 511 Z-planes, 0.5 m apart, scale bar 100 m. Courtesy of Mikhail
Slipchenko, Purdue University.
Feat Olympus_Layout 1 8/30/12 10:00 AM Page 21
different frequencies to serve as pump and
Stokes beams, with the difference in fre-
quency tuned to match the resonant fre-
quency of a specific chemical bond. Re-
cently, however, Adrian Pegoraro et al
have developed a femto-CARS system
that uses the same femtosecond-pulsed IR
laser that is used for multiphoton excita-
tion and splits the beam into two compo-
nents. One component passes through a
photonic crystal fiber to produce a red-
shifted Stokes beam, which then recom-
bines with the unaltered beam using preci-
sion alignments and stage delay.
The CARS signal can be collected in ei-
ther the reflected or forward direction with
qualitatively different patterns, adding in-
formation for expert interpretation. In all
detection modes, the IR wavelengths of
the imaging beams are blocked, and filters
are chosen to separate different signals ac-
cording to wavelength range. For example,
the Olympus femto-CARS system uses
two excitation wavelengths for imaging:
800 and 1040 nm. The 800-nm wave-
length serves as the imaging wavelength
for intrinsic fluorescence and SHG, and is
one of the wavelengths for CARS imag-
ing; 1040 nm functions as the second nec-
essary wavelength for CARS imaging. The
SHG signal from the 800-nm beam occurs
at 400 nm, and the CARS signal is col-
lected at 650 nm.
Depending upon the sample, these three
signals (SHG, CARS and intrinsic fluores-
cence) can be collected simultaneously
using either reflected or forward detection.
Additional wavelengths may be used for
a fourth signal. A four-channel filter set
for the reflected detector might have filters
that collect narrow bands for simultaneous
imaging of SHG; green fluorescent protein
or intrinsic fluorescence; yellow fluores-
cent protein or the DiI fluorescent indica-
tor; and CARS. Because these imaging
modalities are not as specific as antibody-
or nucleic-acid-sequence-recognizing
probes, most experiments are comparative.
For example, the collagen distribution
around a tumor may be compared to unaf-
fected stromal tissue or to the pattern ob-
served in a control animal. Researchers
might, for example, do a comparison be-
tween the intrinsic fluorescence of NADH
under normoxic and hypoxic conditions.
Metabolism may also be expressed as a
ratio of NADH (or NADPH) with another
member of the electron transport chain,
such as flavin adenine dinucleotide. Simi-
larly, the CARS signal indicates lipids in
general but cannot distinguish sphin-
gomyelin from phosphatidylcholine or any
other molecule with a long chain of C-H
bonds.
SHG imaging of collagen
Collagen deposition around tumors has
been recognized by pathologists for
decades. Paolo Provenzano et al have pro-
posed a classification of tumor-associated
collagen signatures that have potential for
clinical staging. These changes in collagen
structure can be observed using SHG im-
aging. Degradation of collagen resulting
from extracellular protease activity, as
well as changes in fibril composition, may
be helpful in identifying changes in tissue
that indicate tumor initiation or progres-
sion.
Combined with collagen imaging,
CARS imaging of lipids has the potential
to help investigate the role of fat-contain-
ing adipocytes in the tumor microenviron-
ment. The observation that cancers appear
to have an association with fat cells might
suggest that adipokines (hormones se-
creted from adipocytes) promote tumori-
genesis by inducing the expression of
genes regulating cancer cell proliferation,
invasion, survival and angiogenesis (Fig-
ure 2). Longitudinal studies may reveal
changes in lipid content and fibrosis as the
tumor progresses.
Intrinsic fluorescence and CARS
The impact of dietary fats on lipid ab-
sorption can be studied using CARS. Indi-
vidual cells in intestinal microvilli are
visualized by their intrinsic fluorescence
in the green wavelengths, while the CARS
signal, detected in the red wavelengths,
highlights lipids both in the lumen of the
gut and in microvillus cells (Figures 3 and
4). Such experiments may help in the de-
sign of therapeutic interventions for meta-
bolic syndrome and other lipid absorption
abnormalities.
More applications
of multimodal imaging
In cancer research, recognizing the im-
portance of the tumor cell microenviron-
ment has led to an increase in studies in
living animals and animal tissue. Multi-
modal imaging provides a way to eluci-
date the relationship between obesity and
cancer using CARS imaging of lipids, the
mechanisms of invasion and metastasis
using SHG, and altered metabolism using
intrinsic fluorescence. Epidemiological
studies reveal a correlation between cer-
tain cancers and obesity, but finding a di-
rect cause-and-effect relationship has
proved elusive.
In a series of studies using CARS, Ji-
Xin Cheng, Thuc T. Le and colleagues ap-
proached this question using CARS imag-
ing of lipids in cells and in animal models.
They observed increased lung metastasis
and circulating tumor cells in animals fed
a high-fat diet. Microscopic imaging of
these cells showed increased lipid deposi-
tion compared to tumor cells from animals
fed a low-fat diet.
In neuroscience, significant research is
being conducted with the ultimate goal of
alleviating myelin degradation in spinal
cord injury and demyelinating diseases
such as multiple sclerosis. CARS imaging
of myelin sheath, SHG imaging of as-
troglial processes and two-photon excita-
tion fluorescence imaging of calcium ion
distribution in live spinal tissues may help
researchers identify opportunities for inter-
vention.
In animal models of cardiovascular dis-
ease, the stability of atherosclerotic plaque
can be assessed using multimodal label-
free imaging. Lipid-rich macrophages
known as foam cells are increased in un-
stable plaque, while underlying collagen
and elastin are disrupted. CARS can iden-
tify the lipid content of the accumulated
macrophages, while SHG indicates disor-
22 BioPhotonics September 2012
Multimodal Label-Free Imaging
Figure 4. Imaging of lipids within cells of the
microvillus, with CARS signal (magenta) from
lipids and intrinsic fluorescence (green) delineating
individual cells of the microvillus. Maximum intensity
projection, 495 Z-planes, 0.3 m apart with
calculated orthogonal slices. Courtesy of Mikhail
Slipchenko, Purdue University.
Feat Olympus_Layout 1 8/30/12 10:00 AM Page 22
ganized collagen fibrils; the intrinsic fluo-
rescence of elastin can highlight the vul-
nerability of the lesion.
Multimodal label-free imaging offers a
way to monitor disease processes over
time with less physiological perturbation.
Now that there are turnkey multimodal
systems available, biomedical researchers
can combine these methodologies for a
more thorough understanding of disease
processes. Moreover, these microscopic
imaging techniques can pave the way for
developing specific molecular imaging
techniques that offer greater tissue depth
and even the potential for clinical utility.
Meet the authors
James Lopez is the confocal and multiphoton
sales application specialist at Olympus America
Inc. in Chicago; email: james.lopez@olympus.
com. Yiwei Jia is marketing manager of confo-
cal laser scanning microscopes at Olympus
America Inc. in Center Valley, Pa.; email:
yiwei.jia@olympus.com.
References
K.A. Kasischke et al (July 2, 2004). Neural ac-
tivity triggers neuronal oxidative metabolism
followed by astrocytic glycolysis. Science,
pp. 99-103.
T.T. Le et al (January 2009). Coherent anti-
Stokes Raman scattering imaging of lipids
in cancer metastasis. BMC Cancer, Vol. 9,
p. 42.
T.T. Le et al (2010). Shedding new light on
lipid biology with coherent anti-Stokes
Raman scattering microscopy. J Lipid Res,
Vol. 51, p. 3091.
A.F. Pegoraro et al (February 2009). Optimally
chirped multimodal CARS microscopy based
on a single Ti:sapphire oscillator. Opt Ex-
press, Vol. 17, p. 2984.
P.P. Provenzano et al (December 2006). Colla-
gen reorganization at the tumor-stromal in-
terface facilitates local invasion. BMC Med,
Vol. 4, p. 38.
P.P. Provenzano et al (2009). Multiphoton mi-
croscopy and fluorescence lifetime imaging
microscopy (FLIM) to monitor metastasis
and the tumor microenvironment. Clin Exp
Metastasis, Vol. 26, pp. 357-370.
P.P. Provenzano et al (November 2009). Shin-
ing new light on 3D cell motility and the
metastatic process. Trends Cell Biology,
Vol. 19, pp. 638-648.
P.P. Provenzano et al (December 2009). Matrix
density-induced mechanoregulation of breast
cell phenotype, signaling and gene expres-
sion through a FAK-ERK linkage. Onco-
gene, Vol. 28, pp. 4326-4343.
M. Skala and N. Ramanujam (2010). Multipho-
ton redox ratio imaging for metabolic moni-
toring in vivo. Methods Mol Biol, Vol. 594,
pp. 155-162.
M.C. Skala et al (December 2007). In vivo
multiphoton microscopy of NADH and
FAD redox states, fluorescence lifetimes,
and cellular morphology in precancerous
epithelia. Proc Natl Acad Sci, Vol. 104, pp.
19494-19499.
This article is the final installment in
a series. The first three articles in this
series are:
Adrian Pegoraro et al (October 2009).
CARS Microscopy Made Simple.
BioPhotonics.
Angela Goodacre et al (October 2010).
Combining Second-Harmonic Generation
with Multiphoton Imaging. BioPhotonics.
Angela Goodacre and Dennis Donley
(September 2011). Intrinsic Fluorescence
Lights Up Cellular Components.
BioPhotonics.
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