Biophotonics201311 Dl

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NOVEMBER 2013

www.Photonics.com

LIVE-CELLIMAGING

EVOLVESTOFINDNEWNICHES
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4 BioPhotonics November 2013

PHOTONICS

The technology of generating and harnessing light and other forms of radiant energy whosequantum 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.

10BIOSCAN BioPhotonicseditors curate the most significant headlines

of the month for photonics in the life sciences and take you

deeper inside the news. Featured stories include:

Sculpted light captures brain activity

Laser-based tool tells normal tissue from tumors

Light to the heart to restore healthy beats

17RAPIDSCAN Femtosecond lasers seeing huge growth for cataract surgery

www.photonics.com

Volume 20 Issue 8

23LIVE-CELL IMAGING EVOLVES TO FIND NEW NICHES by Marie Freebody, Contributing Editor

Both bright-eld and uorescence techniques can capture

cellular processes for observation of real-time dynamics.

27MOVING PAST THE ARTICULATED ARM by Gary Boas, News Editor

Fiber lasers offer surgical applications more versatility and

exibility, and easier integration into medical instruments.

30Q&A: ADAPTIVE OPTICS ON THE RISE by Laura S. Marshall, Managing Editor

Three company representatives touch on the variables driving

the market for adaptive optics for biological applications.

33THE RECENT HISTORY OF ENDOSCOPE DESIGN:WAY MORE THAN CANDLELIGHT AND SPECULA

by Gary Boas, News Editor

Endoscope designs have evolved over the past several years

to provide clinicians a better view of the gastrointestinal tract.

8EDITORIAL

37BREAKTHROUGHPRODUCTS

40APPOINTMENTS Upcoming Courses and Shows

41ADVERTISER INDEX

42POST SCRIPTS by Caren B. Les

Buttery wings + nanotubes = new optical material

NEWS

FEATURES

DEPARTMENTS

30

THE COVER

Live-cell imaging of (left,

top to bottom) Tetrahymena,

fibroblast mitochondria,

hippocampal neuron; and

(right) cancer cell mitochon-

dria. Design by Art Director

Suzanne L. Schmidt.

Now available as a

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for subscribers

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BioPhotonics November 2013

Group Publisher Karen A. Newman

Editorial Staff

Managing Editor Laura S. MarshallSenior Editor Melinda A. Rose

News Editors Gary Boas, Caren B. LesContributing Editors Hank Hogan, Marie Freebody

Copy Editors Judith E. Storie, Margaret W. Bushee, Christopher Goodell

Creative Staff

Senior Art Director Lisa N. ComstockBioPhotonics Art Director Suzanne L. Schmidt

Designer Janice R. TynanDirector Charley Rose

Multimedia Services & Marketing

Director of Publishing Operations Kathleen A. Alibozek

Electronic Media Staff

Web Development Team Leader Brian L. LeMireWeb Developers Alan W. Shepherd

Brian A. Bilodeau

Corporate Staff

Chairman/Founder Teddi C. LaurinPresident Thomas F. Laurin

Vice President Kristina A. LaurinVice President Ryan F. Laurin

Controller Mollie M. ArmstrongAccounting Manager Lynne Lemanski

Accounts Receivable Manager Kathleen G. PaczosaBusiness Manager Elaine M. Filiault

Human Resources Coordinator Carol J. AtwaterAdministrative Assistant Marge Rivard

Business Staff

Director of Sales Ken TyburskiAssociate Director Rebecca L. Pontier

Trade Show Coordinator T. Dylan AcostaComputer Assistant Angel L. MartinezCirculation Manager Heidi L. Miller

Assistant Circulation Manager Melissa J. LiebenowCirculation Assistants Alice M. White, Kimberly M. LaFleur,

Theresa A. HornSubscriptions Janice L. Butler

Traffic Manager Daniel P. Weslowski

Subscription Policy BioPhotonics ISSN-1081-8693 (USPS 013913) is published 9 times per year byLaurin Publishing Co. Inc., 100 West Street, Pittsfield, MA 01201. TITLE reg. in US Library of Congress.The issues will be as follows: January, February/March, April, May/June, July/August, Septem-ber, October, November and December. Copyright 2013 by Laurin Publishing Co. Inc. All rights re-served. POSTMASTER: Periodicals postage paid at Pittsfield, MA, and at additional mailing offices.Postmaster: Send form 3579 to BioPhotonics, 100 West Street, PO Box 4949, Pittsfield, MA 01202-4949, +1(413) 499-0514. CIRCULATION POLICY: BioPhotonics is distributed without charge to qualified research-ers, engineers, practitioners, technicians and management personnel working with the fields of medi-cine or biotechnology. Eligibility requests must be returned with your business card or organizationsletterhead. Rates for others as follows: $45 domestic and $56.25 outside US per year prepaid. Overseaspostage: $30 airmail per year. Publisher reserves the right to refuse nonqualified subscriptions. ARTI-CLES FOR PUBLICATION: Individuals wishing to submit an article for possible publication in BioPhoton-ics should contact Laurin Publishing Co. Inc., 100 West Street, PO Box 4949, Pittsfield, MA 01202-4949;phone: +1 (413) 499-0514; fax: +1 (413) 442-3180; email: [email protected] statementsand opinions expressed in BioPhotonics are those of the contributors the publisher assumes no

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Journeys of Discovery

Little more than 100 years ago, French biologist Jean Comandon and colleagues were

making live lms of cell division using t ime-lapse microcinematography, capturing

the wonder of the process before the dyes and preservatives killed the cells. Fin,indeed. But live-cell microcinematography was the celluloid precursor to modern-day

live-cell imaging. And while the microscope has several hundred years of history over

video capture of dividing cells, the irrepressible urge to see more has brought about

advances in both technologies, and united the two for ever-more-amazing results.

In our cover story, Live-Cell Imaging Evolves to Find New Niches, contribuing

editor Marie Freebody discusses the improvements in microscope technology and

automation that are continually broadening the kinds of cel ls and cellular processes that

can be studied. Challenges persist, however, including controlling light keeping it from

damaging samples and inuencing research results. Read the entire feature beginning on

page23.

Also in th is issue, Managing Editor Laura S. Marshall contributes a Q&A with expertsfrom three companies in the adaptive optics market. Among the take-aways from Q&A:

Adaptive Optics On the Rise is this quote from Christian Theriault, president and

CEO of Tag Optics Inc. of Princeton, N.J.: Unlocking crucial in situ information from

a multitude of depths is where adaptive optics technologies provide a key capability that

may help enable future researchers and clinicians to see phenomena they could never

observe before.

Where is the adaptive optics market for bio applications headed? Michael Feinberg,

director of sales and marketing at Boston Micromachines Corp. in Boston, thinks the

market will shif t away from the lab-built atmosphere that has persisted for more than

10 years to a more OEM instrument-type market. James Joubert, applications scientist

at Photometrics in Tucson, Ariz., predicts that adaptive optics of the future will require

on-the-y analysis of and corrections for changes in the imaging environment. Read

more of these experts thoughts on the adaptive optics market beginning on page30.

One Last ObservationIn May 2011, I wrote about a gentleman named Frank H. Andres, who toiled in his

home lab, observing organisms at work through the eyepiece of his microscope. (See

Wanted: Good Observers atwww.photonics.com/a47080.) Frank contacted me follow-

ing a column I wrote about puzzle-solving bees and their lessons to young students about

the coolness of science (www.photonics.com/a45994). As a good observer himself,

Frank had some research that he felt was promising enough to pass along to some capable

young minds who could further his observations.

Late last month, one of Franks daughters wrote to tell me that her dad had passed

away in September. In her brief note, she underscored the importance he put on explora-

tion, curiosity and observation. Frank told me in 2011 that he had been on a journey ofdiscovery his entire life, and his story continues to inspire me. I hope it inspires you, too.

EDITORIAL

Karen A. [email protected]

BioPhotonicsEditorial Advisory Board

Mark A. Anastasio, Ph.D.

Professor of Biomedical Engineering

Washington University in St. Louis

Stephen A. Boppart, M.D., Ph.D.

Bliss Professor of Engineering

Electrical and Computer

Engineering, Bioengineering and Medicine

Beckman Institute for Advanced

Science and Technology

University of Illinois at Urbana-Champaign

David Benaron, M.D.

Professor, Medicine (consulting)

Founder, Stanford Biomedical Optics program

Stanford University School of Medicine

CEO, Spectros Corp.

Aydogan Ozcan, Ph.D.

Associate Professor

Electrical Engineering, Bioengineering

University of California, Los Angeles

Adam Wax, Ph.D.

Theodore Kennedy Associate Professor

Director of Masters Studies at the Depar tment

of Biomedical Engineering, Duke University

Chairman and Founder, Oncoscope Inc.
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Welcome to

The online companion to BioPhotonicsmagazine

CONTRIBUTORS

News editor Gary Boashas

extensive experience as

a writer and editor in the

research community; he is

also a contributing editor to

Photonics Spectra. Page27

and page 33.

Contributing editor Marie

Freebodyis a freelance jour-

nalist with a masters degree

in physics from the University

of Surrey, England. Page23.

Welcome to

The online companion to BioPhotonicsmagazine

Managing Editor Laura S.

Marshall combines years in

journalism with a lifelong love

of science to cover the vast

world of photonics; in addition

to her magazine duties, she

co-hosts the Light Matters

Weekly Newscast on

Photonics.com.Page30.

9BioPhotonics November 2013

Whats Online:

Log in with your email address

or your magazine subscriber number:

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or call the circulation department

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To download the app,

scan this QR code or visit

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Check out our mobile apps

Our collection of helpful resources for students, educators and researchers, including the

Photonics Dictionary+; Photonics Handbook; a list of societies, associations, universities

and research centers; interactive laser charts; webinars; white papers; and our Light Matters

weekly newscasts.

See the Latest Videos

posted toPhotonics.com

Latest Technology Videos

Light MattersWeekly Newscast

BioPhotonicsmagazine print and digital

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BioPhotonics ...

In the December issue of

Laser Trends

Imaging Trends

Spectroscopy Trends

Microscopy Trends

Youll also nd all the news that affects your

industry, from market reports to the latest

products and media.

Check out a sample of the digital version of

BioPhotonicsmagazine atwww.photonics.

com/DigitalSample. Its a whole new world of

information for people in the global photonics

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Sculpted light captures brain activityVIENNA A high-speed imaging tech-nique that sculpts the 3-D dist ribution

of light in a sample can resolve a single

neuron in a living worm, opening pos-

sibilities for studying the function of the

organisms nervous system and pairing

brain function to anatomy.

A major aim of neuroscience today is

to understand how an organisms nervous

system processes sensory input and gener-

ates behavior by observing the activity

of cells across the entire brain. But to

do this, scientists need detailed maps

of how the nerve cells are wired in thebrain as well as information on how these

networks interact in real time.

Until now, researchers had focused on

studying the activity of single neurons

and small networks in the C. elegans

worm, but hadnt been able to establish

a functional map of the entire nervous

system because of the limitat ions of the

imaging techniques used. The activity

of single cells can be resolved with high

precision, but simultaneously looking at

the function of all the neurons in an ent ire

brain has been a major challenge, withthe trade-off being between spatial or

temporal accuracy.

Previously, we would have to scan

the focused light by the microscope in

all three dimensions, said quantum

physicist Robert Prevedel, a member of

the University of Vienna team of physi-

cists and neurobiologists that developed

the new technique. That takes far too

long to record the activity of al l neurons

at the same t ime. The trick we invented

tinkers with the light waves in a way that

allows us to generate discs of light in the

sample.

That means they have to scan in only

one dimension to get the information they

need, he said. We end up with three-di-

mensional videos that show the simul-

taneous activities of a large number of

neurons and how they change over time.

But microscopy wasnt the only challenge

to imaging the worms brain: Visualizing

the neurons requires tagging them with

a fuorescent protein that lights up when

it binds to calcium, signaling the nerve

cells activity.

The neurons in a worms head are

BIOSCAN

A closer look at the most significant biophotonics research and technology headlines of the month

Frontal part of a nematode seen through a microscope: The neurons of the worms brain are coloredin green. Above is an artists interpretation of the discs of light generated by the WF-TeFo (wide-field

temporal focusing) microscope, scanning the brain area and recording the activity of certain neurons.Photos courtesy of IMP.

Artists rendering of a differential interference contrast microscope image overlaid with neurons in thehead ganglia of the nematode C. elegans. Discs indicate the sculpted-light excitation scheme used for

high-speed functional imaging of neural dynamics.


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so densely packed that we could notdistinguish them on our rst images, said

neurobiologist Tina Schrdel, a doctoral

candidate in the lab of Research Institute

of Molecular Pathology (IMP) group

leader Manuel Zimmer. Our solution was

to insert the calcium sensor into the nuclei

rather than the entire cells, thereby sharp-

ening the image so we could identify sin-

gle neurons. Schrdel is co-rst author ofa study on the work published inNature

Methods (doi: 10.1038/nmeth.2637).

The researchers recorded 70 percent

of the nerve cells activity in the worms

head with high spatial and temporal reso-

lution, which could enable experiments

not possible before. One question that will

be addressed is how the brain processes

sensory information to plan and executespecic movements.

Answering that will require further

renement of both the microscopy and

computational methods to study freely

moving animals, the team members say,

something they hope to achieve in the

next two years.

Laser-based tool tells normal tissue from tumors

ANN ARBOR, Mich., and CAMBRIDGE,Mass. A new laser tool can microscopi-

cally distinguish between normal and

cancerous brain tissue in real time. It

doesnt miss cells that could trigger new

tumor growth, so it could make brain

cancer surgery much more effective.

The approach, called SRS (stimulated

Raman scat tering) microscopy, was

developed and tested by a multidisci-

plinary team of chemists, neurosurgeons,

pathologists and others afliated with the

University of Michigan Medical School

and Harvard University.This is the rst t ime SRS microscopy

has been used in a living organism to

see tumor margins the boundary area

where tumor cells inltrate among normal

ones. Thats the hardest area for surgeons

to tackle, especially when a tumor has

invaded a region with an important neuro-

logical function.

With the SRS technique, they can de-

tect a weak light signal that comes out of

a material after it is hit with light from a

noninvasive laser. By carefully analyzing

the spectrum of colors in the light signal,

the researchers can tell a lot about the

chemical makeup of the sample.

Over the past 15 years, Harvards Dr.

Sunney Xie, the co-lead author of the

paper, has advanced the technique for

high-speed chemical imaging. By am-

plifying the weak Raman signal by more

than 10,000 times, it is now possible to

make multicolor SRS images of living t is-

sue or other materials. The team can even

make 30 new images every second the

rate needed to create videos of the tissue

in real time.

The authors suggest that SRS micros-

copy may be as accurate for detecting

tumors as the approach currently used in

brain tumor diagnosis, called H&E stain-

ing. Comparing the two approaches, three

surgical pathologists had nearly the same

level of accuracy, no matter which images

they studied. But unlike H&E staining,

SRS microscopy can be done in real time,

and without dyeing, removing or process-

ing the tissue.

The team used the tool to see the tini-

est areas of a human glioblastoma brain

tumor in a live mouses brain tissue. They

imaged tissue removed from a patient

with gl ioblastoma multiforme, one of the

most deadly brain tumors. Surgery is one

of the most effective treatments, but less

than one-fourth of operations achieve the

best possible results, according to a study

published last fall in the Journal of Neuro-

surgery.

Though brain tumor surgery has ad-

vanced in many ways, survival for many

patients is still poor, in part because sur-

This image of a human glioblastoma brain tumor in the brain of a mouse was made with stimulatedRaman scattering microscopy. The technique allows the tumor (blue) to be easily distinguished fromnormal tissue (green) based on faint signals emitted by tissue with different cellular structures.Courtesy of Xie lab, Harvard University.
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geons cant be sure that theyve removed

all tumor tissue before the operation is

over, said co-lead author Daniel Orrin-

ger, M.D., a lecturer in the U-M depart-

ment of neurosurgery.

Biopsy has been the gold standard for

detecting and removing these types oftumors, Xie told theHarvard Gazette.

But this technique, we believe, is better

because its live. Surgeons can now skip

all the steps of taking a biopsy, freezing

and staining the tissue. This technique

allows them to do it al l in vivo.

The work was featured in Science

Translational Medicine(doi: 10.1126/sci-

translmed.3005954).Currently, the SRS microscopy system

is not small or stable enough to be used in

an operating room. The team is collabo-

rating with a startup company formed by

members of Xies group, called Invenio

Imaging Inc., which is developing a laser

to perform SRS through inexpensive ber

optic components. The team also is work-

ing with AdvancedMEMS Inc. to reducethe size of the probe that makes the im-

ages possible.

Connect with the leading mindsin the imaging community

Register Todaywww.spie.org/mi1

Conference & Courses

1520 February 2014

LocationTown & Country Resortand Convention Center

San Diego, California, USA

2014

BIOSCANb

Light to the heart to restore healthy beatsBALTIMORE When a persons heart

slows or stops, the current practice is to

jump-star t it with a blast of elect ricity

from a pacemaker or debrillator. But

a multiuniversity team aims to put an

optogenetic twist on the procedure by re-

placing the violent jolt of electricity withgently applied light.

Applying electricity to the heart has

its drawbacks, said Natalia Trayanova,

a professor at Johns Hopkins University

and team leader of a group of biomedi-

cal engineers from Johns Hopkins and

Stony Brook University. When we use a

debrillator, its like blasting open a door

because we dont have the key. It applies

too much force and too little nesse. We

want to control this treatment in a more

intelligent way. We think its possible to

use light to reshape the behavior of the

heart without blasting it.In optogenetics, light-sensitive proteins

called opsins are already being inserted

into cells to control certain brain activi-

ties. When exposed to light, these proteins

become tiny portals within the target

cells, allowing a stream of ions to pass

through. Early researchers have begun

using this tactic to control the bioelectric

behavior of certain brain cells, forming

a rst step toward treating psychiatric

disorders with light.

The researchers plan to give the tech-

nique a cardiac twist so that, in the near

future, doctors can use low-energy light

to solve serious heart problems such as

arrhythmia. They plan to accomplish theirless-painful method by using biological

lab data and intricate computer modeling.

Trayanova has spent many years devel-

oping highly detailed computer models

of the heart, simulating whole cardiac

behavior as well as molecular and cellular

behavior. The researchers report that they

have successfully tested the light-based

tactic on the computer-modeled heart.

The Johns Hopkins researchers will use

the model to conduct virtual experiments,

trying to determine how to position and

control the light-sensitive cells to help the

heart maintain healthy rhythm and pump-

ing activity. They also will try to gauge

how much light is needed to activate the

healing process.

Collaborators at Stony Brook are work-

ing on techniques to make heart t issue

light-sensitive by inserting opsins into

cells. They also will test how these cells

respond when illuminated. The goal is to

use the computer model to move closer to

the day when doctors can begin treat-

ing their heart patients with gentle light

beams. The researchers say it could hap-

pen within a decade.

Natalia Trayanova
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The most promising thing about hav-

ing a digital framework that is so accurate

and reliable is that we can anticipate

which experiments are really worth do-ing to move this technology along more

quickly, said postdoctoral fellow Patrick

M. Boyle. One of the great things about

using light is that it can be di rected at

very specic areas. It also involves very

little energy. In many cases, its less

harmful and more efcient than

electricity.

The research was published inNature

Communications(doi: 10.1038/ncom-

ms3370).After the technology is honedthrough the computer modeling tests, it

could be incorporated into l ight-based

pacemakers and debrillators.

BIOSCAN b

In this illustration, the optrode at left delivers blue light to the heart via a fiber optic tip. In the enlarge-ment at right, a heart cell (large red oval) contains an implanted light-sensitive opsin (blue oval) that worksalongside the hearts own proteins (yellow ovals). This teamwork allows the cell to convert light energy intoan electric kick to trigger a healthy heartbeat. Courtesy of Patrick M. Boyle.

Superbright nanocrystals advancebiosensingSYDNEY and ADELAIDE, Australia

Superbright, photostable and background-

free nanocrystals called SuperDots

three orders of magnitude brighter than

quantum dots enable a new approach to

highly advanced biosensing technologies

using optical bers. When combined with

a unique optical ber that a llows light to

interact with nanoscale volumes of liquid,

SuperDots al low a single nanopart icle to

be detected from a distance.

The implications for nanoscale applica-

tions such as biodetection and bioimaging

are signicant, according to the research-

ers who developed the technique. Up

until now, measuring a single nanopar-

ticle would have required placing it inside

a very bulky and expensive microscope,

said professor Tanya Monro, director of

the University of Adelaides Institute for

Photonics and Advanced Sensing (IPAS)

and a member of the team that made the

discovery with colleagues from Macqua-

rie University in Sydney and Peking Uni-

versity in China. For the rst time, weve

been able to detect a single nanoparticle at

one end of an optical ber from the other

end. That opens up all sorts of possibili-

ties in sensing.

Nanocrystals can be doped with sensi-

tizer ions that absorb infrared radiation,

then transfer their excitation to activa-
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BioPhotonics November 2013

tor ions that then emit visible light. Themore dopants that are added, the higher

the emission brightness but only up to a

point. At a certain relatively low thresh-

old, activator ions are maxed out, and

brightness begins to diminish.

The team found that high levels of

infrared radiation combined with higher

activator concentrations in excess of the

threshold led to signicantly enhanced

luminescence signals by up to a factor

of 70, an improvement of three orders of

magnitude over quantum dots.

These single nanocrystals were bright

and sensitive enough to be detected re-

motely using an optical ber, or to be seen

with the naked eye through an automated

scanning microscope, so they could

provide high-contrast biolabels to track

individual cells or sense single molecules.

The special optical ber engineered at

IPAS also proved useful in understanding

the properties of nanoparticles.

Material scientists have faced a huge

challenge in increasing the brightness

of nanocrystals, said Dr. Dayong Jin of

Macquaries Advanced Cytometry Labo-

ratories. Using these optical bers, how-

ever, we have been given

unprecedented insight

into the light emissions.

Now, thousands of emit-

ters can be incorporated

into a single SuperDot,

creating a far brighter andmore easily detectable

nanocrystal.

Using optical bers,

we can get to many

places, such as inside the

living human brain, next

to a developing embryo or

within an ar tery loca-

tions that are inaccessible

to conventional measure-

ment tools, Monro said.

This advance ultimately

paves the way to breakthroughs in medi-cal treatment. For example, measuring

a cells reaction in real t ime to a cancer

drug means doctors could tell at the time

treatment is being delivered whether or

not a person is responding to the therapy.

Under infrared illumination, SuperDots

selectively produce bright blue, red and

infrared light with 1000 times more sensi-

tivity than existing materials.

Neither the glass of the optical ber

nor other background biological mole-

cules respond to infrared, so that removed

the background signal issue. By exciting

these SuperDots, we were able to lower

the detection limit to the ultimate level: a

single nanoparticle, Jin said.

The work appears inNature Nanotech-

nology(doi: 10.1038/nnano.2013.171).

Macquarie is working with industry

partners Minomic International Ltd. and

Patrys Ltd. to develop uses for SuperDots

in cancer diagnostic kits. The university

is now seeking other industrial partners

with the capacity to jointly develop solu-

tions outside of these elds.

BIOSCANb

SuperDots enable the microstructured optical fiber to detect and trackthe movement of a single nanocrystal remotely. Courtesy ofDr. Mathieu Juan.

The microstructured optical fiber has been employed as a nanoliter-volume spectroscope to analyze theoptical properties of nanocrystals. Courtesy of Matthew Henderson.
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ZURICH, and LIVERMORE, Calif.

Spaghett ilike arrays of gold-coated

metallic carbon nanotubes can amplify

the signals of surface-enhanced Raman

spectroscopy (SERS) enough to allow the

performance of analyses that are more

reliable, sensitive and cost-effective.

Potential applications include real-time

point-of-care monitoring of physiological

levels, fast screening of drugs and toxins

in law enforcement, and early detection of

biological weapons.

Previously, the detection limit of com-

mon SERS systems was in the nanomolar

range (one billionth of a mole), provid-

ing adequate signal strength in isolated

cases only, and yielding results with

low reproducibility. But a new sensor

developed jointly by ETH Zurich and the

Lawrence Livermore National Laboratory

(LLNL) uses plasmonics to massively am-

plify the signals of Raman-scattered light:

The researchers detected a certain organic

species (1,2-bis(4-pyridyl)ethylene, or

BPE) in a concentration of a few hundred

femtomoles per liter (a 100-femtomolar

solution contains around 60 mill ion mol-

ecules per liter).

Raman spectroscopy takes advantage

of the fact that molecules illuminated by

xed-frequency light exhibit inelasticscattering closely related to the vibra-

tional and rotational modes excited in

the molecules. Such light differs from

common Rayleigh scattered light in that it

has different frequencies than that of the

irradiating l ight and produces a specic

frequency pattern for each substance

examined, making it possible to use this

spectrum information as a ngerprint

for detecting and identifying specic

substances.

To analyze individual molecules, the

frequency signals must be amplied,which requires that the molecule in ques-

tion either be present in a h igh concen-

tration, or be situated close to a metall ic

surface that amplies the signal (surface-

enhanced Raman spectroscopy).

The substrate used by doctoral student

Ali Altun and professor Hyung Gyu Park,

both of ETH Zurich, and LLNL capability

leader Tiziana Bond was vertically ar-

ranged, caespitose, densely packed carbon

nanotubes (CNTs) that guarantee a high

density of hot spots. The group developedtechniques to grow dense forests of CNTs

in a uniform and controlled manner.

CNT tips are sharply curved and

coated with gold and hafnium dioxide, a

dielectric insulating material. The point

of contact between the sensors surface

and the sample resembles a plate of

spaghetti topped with sauce. But between

the strands of spaghetti are numerous

randomly arranged holes that let scattered

light through, with the many points of

contact amplifying the signals.

One method of making highly sensi-tive SERS sensors is to take advantage of

the contact points of metal nanowires,

Park said. The nanospaghetti structure

with metal-coated CNT tips is perfect for

maximizing the density of these contact

points. The wide distribution of metal-

lic nanocrevices in the nanometer range,

recognized to be responsible for extreme

electromagnetic enhancement, resulted in

intense and reproducible amplication.

The CNTs were coated with the insula-

tor before a layer of gold was applied toprevent plasmonic energy leakage. This

was the breakthrough, Altun said. The

insulation layer increased the sensitiv-

ity of the sensor substrate by a factor of

100,000 in the molar concentration unit.

For us as scientists, this was a moment

of triumph, said Park, and it showed us

that we had made the right hypothesis and

a rational design.

The work appears inAdvanced Materi-

als(doi: 10.1002/adma.201300571). Park

and Bond hope to commercialize the tech-

nology with an industry partner, but wantto improve the sensors sensitivity and

nd potential areas of application.

BIOSCAN b

Spaghettilike surface makes stronger SERS sensor

The high-sensitivity sensor is based on the curved tips of carbon nanotubes. The numerous gaps in thespaghettilike construction let the Raman-scattered light pass through. Courtesy of H.G. Park, ETH Zurich.

Plasmofluidic lens is tunable, reconfigurableUNIVERSITY PARK, Pa. Laser-

induced bubbles on a metal lm are the

rst demonstration of a plasmonic lens in

a microuidic environment. Integrating

plasmonics and microuidics could help

in developing highly sensitive biomedical

detection systems and more.

Plasmonics is promising for these appli-

cations because it enables light manipula-

tion beyond the diffraction limit. Nano-

plasmonics combines the speed of optical

communication with the portability of

electronic circuitry in situations where

conventional optics do not work, although

aiming and focusing are difcult. How-

ever, the majority of plasmonic devices

created to date are solid state and lack the

ability to deliver multiple functions.

There are different solid-state devices

to control [light beams], to switch them

or modulate them, but the tenability and
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recongurability are very limited, said

Tony Jun Huang, associate professor of

engineering science and mechanics at

Pennsylvania State University. Using a

bubble has a lot of advantages.

The main advantage of a bubble lens

is just how quickly and easily its loca-

tion, size and shape can be recongured,

affecting the direction and focus of any

light beam passing through it. The teams

plasmouidic lens a lso doesnt require

sophisticated nanofabrication and uses

only a single low-cost diode laser; the

bubbles themselves are easy to dissolve,

replace and move.

Simply moving the laser or adjusting its

power can change how the bubble will de-

ect a light beam, either as a concentrated

beam at a specic target or as a dispersed

wave. Changing the liquid also affects

how a light beam will refract.

To form the plasmouidic device,

Huangs team used a low-intensity laser

to heat water on a gold surface. Thenanobubbles optical behavior remained

consistent as long as the lasers power and

the environmental temperature stayed

constant.

In addition to its unprecedented re-

congurability and tenability, our bubble

lens has at least one more advantage over

its solid-state counterparts: its natural

smoothness, Huang said. The smoother

the lens is, the better quality of the light

BIOSCANb

A nanoscale light beam modulated by surface plasmon polaritons enters the bubble lens, officially knownas a reconfigurable plasmofluidic lens. The bubble controls the lightwaves, while the grating providesfurther focus. Images courtesy of Tony Jun Huang, Penn State.

Laboratory images of a light beam without a bubble lens, followed by three examples of different bubble lenses altering the light.

that pass through it.

The next step is to nd out how the

bubbles shape inuences the direction of

the light beam and the location of its focal

point. Fine control over these light beams

will lead to improvements for on-chip

biomedical devices and superresolutionimaging. For all these applications, you

really need to precisely control light in na-

noscale, and thats where this work can be

a very important component, Huang said.

In addition to researchers from Penn

State, the work involved collaboration

with Northeastern University and MIT.

The results were published inNature

Communications (doi: 10.1038/ncom-

ms3305).
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Femtosecond lasers seeing huge growth for cataract surgery

The global market for femtosecond

lasers for cataract surgery is grow-

ing exponentially and will reach

$2.4 billion by 2019, according toFem-

tosecond Lasers for Cataract Surgery:

Market Shares, Strategies, and Forecasts,

Worldwide, Nanotechnology, 2013 to 2018,

a new market report from RnR Market

Research of Dallas.

In 2012, the market was valued at

$572 million; the company predicts that

in 2013, that value will hit $1.1 billion.

The drivers for this sudden growth are

new competitors in the market, surgeons

need for greater accuracy in cataract

surgery and increasing patient demand for

the laser technology.

That demand is expected to continuegrowing around the world as the aging

population develops cataracts. The simul-

taneous change in demographics and

the introduction of automated processes

will cause an explosion in demand for

ophthalmologists services over the next

20 years: Patients older than 65 consume

10 times the eye care of patients younger

than 65, creating unprecedented demand

for cataract surgery, RnR reported.

Ultrasonic phacoemulsifcation has

been the standard of care in cataract re-

moval equipment for four decades, and

RnR predicts that it will remain the

dominant lens removal technology in the

near term.

But laser-assisted cataract surgery

using femtosecond lasers and picosecond

lasers promises to raise the standards

of precision and safety to new heights.

Numerous types and styles of intraocu-

lar lenses of varying sizes, along with

attachment mechanisms, complement

the introduction of the femtosecond laser

cataract surgical systems, which enable

reproducible, predictable and improved

clinical outcomes. Through image-guided

visualization and micron-level laser pre-

cision, surgeons using these systems can

better control the surgery process.

RAPIDSCAN Business and Markets

Congressman tours UCF laser facility

Photonics was the focal point at the

University of Central Florida (UCF)

in Orlando when Rep. John L. Mica

paid a visit to the Center for Research and

Education in Optics and Lasers (CREOL).

Among the technologies on display

were a laser used to break apart cancer

particles and a cellphone that analyzes

blood, allowing Mica to witness the in-

roads light-based technologies have made

in the medical feld, among others.

They are performing phenomenal re-

search in our backyard, Mica said. The

work going on at UCF holds tremendous

potential for our workforce, and will

impact and touch nearly every aspect of

our lives.

The National Photonics Initiative

(NPI), launched this spring by a cadre of

photonics organizations, seeks to unite

experts from industry, academia and the

government to advance photonics R&D,

to grow the US economy and to improvenational security. A key part of the effort

is to educate members of Congress on the

vital role that photonics plays in home-

land security and the US economy.

CREOL Professor Peter Delfyett (left) speaks withRep. John Mica about the University of CentralFloridas work on optical communications.

Courtesy of Karen Norum.

Molecular imaging and targeted therapy are essential tothe successful implementation of precision medicine.

Dr. Hedvig Hricak, Memorial Sloan-Kettering Cancer Center

chairman of radiology and a plenary speaker at the

World Molecular Imaging Societys annual congress


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Avo helps design,produce OptouidicsNanoTweezer

Avo Photonics of Horsham, Pa., a Halma

company, has provided system design and

manufacturing services for Philadelphia-based

Optouidics Inc.s molecular NanoTweezer

system.

The NanoTweezer is made up of three

components: a microscope ow system thatsits atop the microscope platform, removable

waveguide chips inserted into the ow system,

and a benchtop control unit. It allows users to

isolate objects more than 10 times smaller than

those trapped by traditional optical tweezers.

Avo provided the design and production

of disposable assemblies consisting of three

single-mode, polarization-maintaining bers

hard-coupled to a NanoTweezer chip. Avo also

created the means to tune the wavelength and

control the power of the laser.

Zecotek Photonics raises $3.5MZecotek Photonics Inc. of Vancouver, British Columbia, Canada, has sold more than

5.9 million shares of company stock at a price of 58 cents a share to raise approxi-mately $3.5 million.

The sale increases the nonbrokered private placement the company announced in

August by an additional $260,000 to a total of $3,460,824. Zecotek said it will use the

funds to complete the transfer of technology for commercialization, strengthen its IP

portfolio, and nance purchase orders and general working capital needs. In June, the

company announced that it had raised $2.4 million.

Founded in 2004, Zecotek Photonics develops scintillat ion crystals, photodetectors,

positron emission tomography scanning technologies, 3-D autostereoscopic displays,

and lasers for medical, high-tech and industrial applications under three divisions:

Imaging Systems, Laser Systems and 3-D Display Systems.

In other Zecotek news, Hamamatsu Photonics of Tokyo has placed a $500,000 order

with Zecotek for scintillation crystals to be used in third-party positron emission

tomography (PET) scanners. This is the rst order since Julys announcement of ajoint collaboration agreement between Singapore-based subsidiary Zecotek Imaging

Systems Pte Ltd. and optoelectronic components supplier Hamamatsu Photonics KK.

Bruker acquiresPrairie TechnologiesScientic instruments provider Bruker

Corp. of Billerica, Mass., has acquired

Prairie Technologies Inc., a provider

of uorescence microscopy products.Specic terms were not disclosed.

Madison, Wis.-based Prairie generated

revenues of approximately $11 million in

2012 and has approximately 30 employees

globally. It offers a variety of life sciences

products, including two-photon micro-

scopes; multipoint scanning confocal

microscopes; laser illumination sources;

photoactivation, photostimulation and

photoablation accessories; and synchroni-

zation and analysis software. Applications

include uncaging experiments, optogenet-

ics, electrophysiology studies and cell

biology.

The purchase allows Bruker to enter

the uorescence microscopy market and

strengthens the companys life sciences

offerings by adding to its Nano Surfaces

Divisions existing life sciences atomic

force microscopy (Bio-AFM) systems.

Urologists need better informationon laser ber diameterThe advertised total and core diameters of laser fibers for urological surgery

do not correspond with the actual diameters, according to a new study.Furthermore, there are serious differences between manufacturers of fiberswith supposedly equal diameters.

Urologists need to know the exact technical specifications of the materialthey use, said Dr. Peter Kronenberg of Hospital Fernando Fonseca in Amado-ra, Portugal, who conducted the study with professor Olivier Traxer of HpitalTenon in Paris. If the information conveyed to them, whether written on aproduct label or transmitted by an industry representative, is incorrect, theirjudgments and the decisions they make based on this knowledge may havesurgical repercussions, Kronenberg said.

They evaluated 14 different laser fibers from six brands with advertiseddiameters of 200, 270, 272, 273, 365 and 400 m, taking multiple measure-ments of both the total diameter (including fiber coating) and the fiber corediameter and comparing them to their respective advertised diameters.

The total and the core diameters measured were both significantly differentfrom the advertised diameter in all fibers (p


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RAPIDSCAN r

Agilent opens spectroscopycenter in AustraliaAgilent Technologies Inc. has opened a $25 million center in

Mulgrave, Australia, for cutting-edge spectroscopy research,

laboratory testing and training.

The facility provides advanced communications technology to

facilitate collaboration amongAgilent divisions, research part-

ners and global customers. In the US, Agilent recently opened a

calibration center in Phoenix.

Spectroscopy instruments determine quality and screen for

contaminants in a variety of applications, including agriculture,

the environment and pharmaceuticals.

One of the interesting paradoxes ofbiomedical innovation is increasingly going

to be that even though we have thescientic knowledge required to provide

potentially better treatments for patients or even to prevent disease in those who

are at high risk we may be unable to helppatients benet from them anytime soon.

research associate Magdalini Papadaki, MIT, who recommends

a new science of collaboration between companies, regulatory

agencies and patient groups to advance biomedical innovation;

Papadaki co-authored a paper on the subject with Gigi Hirsch, a

physician-entrepreneur and executive director of the MIT Center

for Biomedical Innovation; the article was published in Science

Translational Medicine (doi: 10.1126/scitranslmed.3006903)

Verisante completesimager prototypeVerisante Technology Inc. of Vancouver, British Columbia,

Canada, has completed the second phase of a rapid multispec-

tral imaging (MSI) system for skin cancer detection.

The prototype is cur rently undergoing laboratory testing

at the BC Cancer Agency Research Centre before starting in

vivo data collection for training the predictive algorithm for

the device.

Verisante licensed the MSI technology from its inventors,

Dr. Haishan Zeng and Dr. Yasser Fawzy of the BC CancerAgency, for skin cancer and oral cancer detection as part of a

broader acquisition strategy to enhance its intellectual prop-

erty portfolio, which also includes white-light reectance im-

aging, uorescence imaging and rapid Raman spectroscopy.

The company will also explore the possibility of com-

bining the MSI with its existing Aura device to determine

whether the two technologies in tandem produce results with

greater accuracy, since they are measuring different param-

eters. The Aura was awarded a 2013 Prism Award for photon-

ics innovation by Photonics Media and SPIE, and an Edison

Award for Excellence in Innovation in 2013.
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LSO Medical partners withGME on dermatology lasers

LSO Medical SAS of Loos, France, amanufacturer of lasers for aesthetic andsurgical applications, has joined forceswith GME German Medical Engineer-ing GmbH of Erlangen, Germany, tomarket GMEs dermatological productline in France. The first products it willmarket are DotScan 10 600, a fraction-al CO

2laser device for the treatment

of wrinkles and scars, and the LinScan808, a diode laser for hair removal.

$18.9 billion the global market value that

biosensors will reach by 2018, according

to a market report f rom TransparencyMarket Research; in 2011, the global

biosensors market value was $9.9 billion,

and it is expected to grow

at a CAGR of 9.6% from 2012 to 2018

The world of biophotonics is constantlyevolving and rightly so, as new technol-

ogies and different applications for exist-

ing devices come into play. Ten years

ago, researchers were starting to explore

photonics-based methods of analyzing

fetal cells or DNA in the mothers blood-

stream as a way to monitor fetal health.

2003Its fnicky. Sometimes

theres no diagnosis.So it is still at the

investigational stage. Dennis Lo, professor of chemical pathology

at the Chinese University of Hong Kong
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Fibercore of Southampton, England, has appoint-

ed John Leesenior vice president of global sales.Prior to joining Fibercore, Lee headed sales and

marketing at Timbercon, where he oversaw 25

percent compound annual growth over a nine-year

period. Other notable roles include co-founding

and heading up sales and marketing for Zmation,

a manufacturer of custom automated systems,

and leading sales for an automated system line

at KDT Systems. Lee has business development

experience in diverse domestic and international

markets including: defense/aerospace, semicon-

ductor, electronics, alternative energy, oil and gas,

telecommunications, and medical devices.

Imaging technology

expert Dr. Roman

Kuranovhas joinedWasatch Photonics

Inc. of Logan, Utah, as

its principal scientist

to oversee develop-

ment of the companys

OCT imaging systems.

Kuranov previously

was a research faculty

member in the depart-

ment of ophthalmol-

ogy in the University of

Texas Health Science

Center at San Antonio, and R&D engineer at

Volcano Corp. and CardioSpectra.

Wasatch Photonics designs, manufactures and

markets high-performance Raman spectrometers,OCT systems, enhanced holographic optics for

optical networking, spectroscopy, test and mea-

surement, and medical imaging applications.

Optoelectronics

industry veteran

Mark Vosloohas

been appointed CEO

of spectrometer

maker Edinburgh

Instruments Ltd.

(EI) of Livingston,

Scotland. Vosloo

joins EI from Oxford

Instruments, where

he held various

management and

senior commercial

roles for the past nine years. Before that, he

worked in the optoelectronics industry for com-

panies including Linos Photonics Ltd. and Horiba

Jobin Yvon Ltd.

Dymax Corp. of Torrington, Conn., has appointed

Andrew Zimsenas a territory sales manager

in the Field Sales Department. Zimsen will help

manufacturers in California solve application

problems and reduce manufacturing costs. Prior

to joining Dymax, Zimsen was the western region

sales manager for ThermoTek Inc. Dymax devel-

ops oligomer, adhesive, coating, dispensing and

light-curing systems for medical devices and other

applications.

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RAPIDSCAN r

Agilent acquires ABC Instrumentacin AnalticaAgilent Technologies Inc. of Santa Clara, Calif., has acquired ABC Instrumentacin

Analtica (ABCIA) of Mexico City. Financial terms were not disclosed; the acquisi-tion was expected to be completed on Oct. 1.

ABCIA is a distributor of analyt ical solutions, including Agilents chemical analy-

sis and life sciences products such as gas and liquid chromatographs, mass spectrom-

eters, atomic and molecular spectroscopy systems, and bioanalyzers.

About 30 ABCIA employees are expected to transfer to Agilent when the acquisi-

tion is nal.

PEOPLEIN THE NEWS

Holomic, ThyroMetrix team up for smartphone labHolomic LLC, the mobile health company founded by UCLA professor Dr. Aydogan

Ozcan, has partnered with clinical thyroid testing company ThyroMetrix USA Inc. of

Williamsburg, Va., to boost products from both companies: The Holomic Rapid Diagnos-

tic Reader (HRDR-200) for smartphones now works in sync with ThyroTest, a thyroid-

stimulating hormone (TSH) blood test available in the US and Canada from ThyroMe-

trix. Point-of-care ofces now will be able to offer instant, low-cost, in-ofce lab results

normally performed and billed by off-premise lab services. ThyroTest is an FDA-regu-

lated and CLIA-waived rapid immunoassay test that enables in-ofce thyroid diagnosis

using a single drop of blood on a small test card. When coupled with the new HRDR-200,

the result is an inexpensive, robust and very portable TSH-testing smartphone lab.
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For more informaon on technical sessions, exhibitors and short courses,

visitwww.picon.org.

March 2-6, 2014Chicago, Illinoiswww.pittcon.org

Follow Usfor special announcements
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Live-Cell ImagingEvolves to Find New NichesFrom watching embryos as they develop to viewing neurons being manipulated

in the brain, live-cell imaging continues to fnd new applications.

BY MARIE FREEBODY, CONTRIBUTING EDITOR

Since its introduction in the 1600s,

improvements in microscope tech-

nology have continually broadened

the types of cells and cellular processes

that can be studied. Advances in automa-

tion have made this already-simple tool

faster and more capable, and time-lapseimaging reveals function and dynamics

in addition to structure.

Live-cell imaging has enabled us to

witness incredible moments in biology

in unprecedented detail. Even embryo-

genesis the process of cell division and

cellular differentiation that occurs at the

earliest stages of life has recently been

captured.

Much of what we know about the cell

cycle, chromosome segregation errors

and the development of cancer has come

from live studies of cells exposed to small

molecule inhibitors and other drugs.

In fact, questions of why diseases take

hold and progress are a major driving

force pushing scientists to improve live-

cell imaging techniques. As well as many

types of cancer, scientists are keen to

unravel the inner workings of neurode-

generative diseases such as Parkinsons

or Alzheimers.

Live-cell imaging can be divided into

bright-eld and uorescence image cap-

ture. Apart f rom documenting changes in

cell shape or cell motion, both technolo-

gies allow for specic and diverse types

of function measurements.

In bright eld, the sample is trans-

illuminated, and the density of the cell

creates contrast that is used to observe

features and dynamics, said Dr. Vytas

Bindokas, facility and technical director

of the Microscopy Core Facility at the

University of Chicago. Bright-eld meth-

ods are among the oldest techniques, but

still highly useful.

Fluorescence methods are extremely

versatile and can t rack multiple specic

types of information at once. By applying

Study of dendritic transport in hippocampal neuron: DIC (bright field) detail is in blue; actin proteinis labeled with red; and an Alzheimers disease-related protein is in green. Courtesy of VirginieBuggia-Prevot and Celia G. Fernandez, Gopal Thinakaran lab.

Fluorescence image of mucocyst tip protein(pink) and DIC bright field (cyan) in Tetrahymena.Moving cells pose a challenge for imaging:Simple cells have long served as models forstudy of similar processes in more complexorganisms. Preparation courtesy of SantoshKumar, Aaron Turkewitz lab.

Vital staining of mitochondria (red) withinfibroblast cells (gray). Dynamics of thesefragile, energy-producing organelles arerecognized as being increasingly importantin disease. Preparation by Sadhana Samant,Mahesh Gupta lab.


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one or more of the ever-growing palette

of uorescent proteins now available, a

bright signal is created against a dark

background.

Vital uorescent dyes in numerous

colors are available to label cellular com-

partments and to read out physiologicprocesses in real time, Bindokas said.

Examples of physiologic measurements

that are associated with major cell events

include changes in pH, ion concentration,

membrane voltage, enzyme activity, gen-

eration of reactive molecules/species, etc.

Superresolutionfor live-cell imaging

One of the biggest trends at the moment

is the t remendous effort under way to get

superresolution techniques working under

live-cell conditions.For many years, it was not possible to

perform any real live-cell experiments

under quasiphysiological conditions,

said Sebastian Tille, director of wide-eld

imaging at Leica Microsystems GmbH

in Germany. This has changed and

delivered new insights. For instance, it

was possible to follow the movement of

dendritic spines with STED [stimulated

emission depletion]. Also, 3-D superreso-

lution (localization) microscopy allows

going far beyond the diffraction limit in

all three spatial dimensions down to themolecular level now possible with the

Leica SR GSD 3D.

Images are now being produced with

detail previously only attained with the

electron microscope. With superresolu-

tion, the limit for visual ization now ap-

proaches the size of common proteins.

Structured illumination microscopy

(SIM) and stochastic optical reconstruc-

tion microscopy (STORM) are two super-

resolution techniques that have advanced

signicantly in recent years, with some

impressive results obtained from living

cells.

SIM uses wide-eld illumination of

the specimen with pat terned excitation to

create interference patterns called moir

fringes. These fringes contain additional

information that can be used to generate a

superresolution image with a twofold im-

provement in resolution over conventional

uorescence techniques.

We are seeing some amazing live-cell

SIM images coming from the labs of Hari

Shroff at the NIH [US National Institutes

of Health] and Eric Betzig of the Howard

Hughes Medical Institute, said Dr. Chris-

Live-Cell Imaging

3-D projections of mitochondria in cancer cells: Recent studies suggest mitochondria may be targetsfor controlling cancer. Preparation by Rifat Hasina, Ravi Salgia lab.

Time-lapse imaging of green fluorescent protein-prion protein in cells expressing the red fluorescentprotein DsRed within the endoplasmic reticulum. Prions can cause neurological diseases that includeKuru and mad cow disease. Preparation from James Mastrianni lab.


25/44


topher OConnell, manager of superreso-

lution system products at Nikon Instru-

ments Inc. in Melville, N.Y. These

researchers are using novel methods of

generating structured illumination to

advance SIM technology to the next level,

creating systems with incredible tempo-

ral resolution and the ability to image atdepth. Even whole embryos can be rapidly

scanned with these methods.

STORM is a form of localization mi-

croscopy that cumulatively maps the uo-

rescence from individual dye molecules

to create an image with ten times better

resolution. Both SIM and STORM are

allowing researchers to observe structural

features that were previously only visible

by electron microscopy.

Other techniquesWhile high-speed confocal imaging

of uorescent reporters seems to be the

most common and powerful tool being

used today for live-cell imaging, there are

other specialized techniques that are used

to study cellular membranes and events.

Total internal reection microscopy, for

example, uses a high-numerical-aperture

objective lens and a spot of laser light to

make the laser reect off the glass-cell

interface and back into the lens. Since the

technique involves shallow illumination,

photodamage to the sample is limited.

A product that has only just reached

the market for whole- or small-organism

Live-Cell Imaging

Expression pattern (intensity color coded) ofBACE1-yellow fluorescent protein in mature hippo-

campal neuron. BACE1 is involved in Alzheimersdisease. Its movement can be tracked from thebulbous cells body, through the thicker dendritesand even the finest axons. Courtesy of VirginieBuggia-Prevot, Gopal Thinakaran lab.

Live-cell STED-CW superresolution of green fluorescent protein-MLCK protein in lung endothelial cells.Region intensity plot shows 39-nm FWHM (full-width half-maximum) diameter for a stress fiber indicatedas ROI1 and green line segment near center image. The bright, upper inset shows the same cell instandard confocal mode. Preparation courtesy of Mary Brown, Steve Dudek lab.

imaging is light-sheet microscopy. This

is similar to a point-scanning confocal

system, except that the scanning is carried

out by a thin sheet of l ight; image capture

is typically at right angles to the plane of

illumination. This is another approach

that causes very litt le photodamage be-

cause it uses high-speed cameras and lowlight, but the labeled cells of the entire

organism can be tracked over time.

Another technique known as spinning-

disc microscopy continues to hold a lot

of power for those looking at monolayer

cells because of the high imaging speed

and resolution that are possible when

using the latest generation of cameras

available on the market.

In this approach, hundreds of spaced

excitation pinholes rotate at high speed

to sweep light across the sample so as

not to interfere with each others signals.

The image can then be captured using

high-sensitivity digital cameras that

include some of the best-efciency photo-

detectors available today (up to 93 percent

efciency).

We recently introduced a new Flash4

sCMOS camera for the UltraVIEW VoX

3-D live-cell imaging system that im-

proves sensitivity and increases frame

rates, said Dr. Jacob Tesdorpf, director of

high-content instruments and applications

at PerkinElmer in Hamburg, Germany.

This enables scientists to capture more

images per second, gaining sensitivity

and speed, and resolving intracellular

process faster than before.

The challengeof imaging the living

Living cells pose a unique challenge

when it comes to microscopy. Firstly, lim-

iting light exposure continues to be oneof the main challenges. Of course, light

is crucial to imaging, but too much light

can manifest in subtle ways to inuence

research results; it can even be damaging

to the cell.

Fluorescence illumination, especially

in the UV range, is harmful for cells and

causes photobleaching and phototoxicity,

Tesdorpf said. It is therefore important

to make every photon count, by limiting

unnecessary excitation to a minimum and

capturing a maximum of emitted photons.

The problem is par ticularly true for

superresolution methods, which require

more raw information than conventional

uorescence methods. This additional

information comes from extra images or

high laser power needed to selectively

turn uorophores on and off.

Apart from light toxicity, mammalian

cells need to be kept at 37 C and may

require elevated levels of carbon dioxide

to maintain vigor.

Various incubation schemes exist as

add-ons to standard microscopes that are

reported to work with varying levels of

success.


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