A portable smart phone-based plasmonic nanosensor readout platform

that measures transmitted light intensities of nanosubstrates using an

ambient light sensor.
Fu Q1, Wu Z, Xu F, Li X, Yao C, Xu M, Sheng L, Yu S, Tang Y.
Author information
Abstract
Plasmonic nanosensors may be used as tools for diagnostic testing in the field of medicine. However,
quantification of plasmonic nanosensors often requires complex and bulky readout instruments. Here, we
report the development of a portable smart phone-based plasmonic nanosensor readout platform (PNRP)
for accurate quantification of plasmonic nanosensors. This device operates by transmitting excitation light
from a LED through a nanosubstrate and measuring the intensity of the transmitted light using the
ambient light sensor of a smart phone. The device is a cylinder with a diameter of 14 mm, a length of 38
mm, and a gross weight of 3.5 g. We demonstrated the utility of this smart phone-based PNRP by
measuring two well-established plasmonic nanosensors with this system. In the first experiment, the
device measured the morphology changes of triangular silver nanoprisms (AgNPRs) in an immunoassay
for the detection of carcinoembryonic antigen (CEA). In the second experiment, the device measured the
aggregation of gold nanoparticles (AuNPs) in an aptamer-based assay for the detection of adenosine
triphosphate (ATP). The results from the smart phone-based PNRP were consistent with those from
commercial spectrophotometers, demonstrating that the smart phone-based PNRP enables accurate
quantification of plasmonic nanosensors.

Smartphone-based system created for disease detection

Posted on November 20, 2017 by bcunning@illinois.edu

Researchers at the University of Illinois at Urbana-Champaign are creating a mobile sensor technology
called PathTracker for performing detection and identification of viral and bacterial pathogens. By
means of a smartphone-based detection instrument, the results are shared with a cloud-based data
management service that will enable physicians to rapidly visualize the geographical and temporal
spread of infectious disease. When deployed by a community of medical users (such as veterinarians or
point-of-care clinicians), the PathTracker system will enable rapid determination and reporting of
instances of infectious disease that can inform treatment and quarantine responses that are currently
not possible with tests performed at central laboratory facilities.

Immediate uses for the technology are for diagnosis of viral infection in human patients (Zika, dengue,
and Chikungunya) and diagnosis of respiratory infection in equine populations (Equine herpesvirus,
Equine influenza, Strangles, Equine adenovirus type 1, and Equine arteritis virus). Further uses include
water testing, food safety, and pharmaceutical quality control.

Leading the research team are Brian Cunningham (PI), Rashid Bashir (Co-PI), and Ian Brooks (Co-PI) from
the University of Illinois and David Hirschberg (Co-PI) from the University of Washington, Tacoma. The
project is supported by the National Science Foundation.
Posted in Uncategorized

Integrated lab-on-a-chip uses smartphone to quickly detect multiple pathogens

Posted on September 15, 2017 by bcunning@illinois.edu

A multidisciplinary group that includes the University of Illinois at Urbana-Champaign and the University
of Washington at Tacoma has developed a novel platform to diagnose infectious disease at the point of
care using a smartphone as the detection instrument in conjunction with a test kit in the format of a
credit card. The group is led by UI Bioengineering and Electrical and Computer Engineering Professor
Brian T. Cunningham, UI Bioengineering Professor Rashid Bashir, and University of Washington at
Tacoma Professor David L. Hirschberg, who is affiliated with the Department of Sciences and
Mathematics, division of the School of Interdisciplinary Arts and Sciences.

Findings have been published in the journal Analytical Chemistry, demonstrating the detection of four
horse respiratory diseases, and in the journal Biomedical Microdevices, for which the system was used
to detect and quantify the presence of Zika, Dengue, and Chikungunya viruses in a droplet of whole
blood. Project collaborators include Dr. David Nash, a private practice equine expert and veterinarian in
Kentucky, and Dr. Ian Brooks, a computer scientist at the National Center for Supercomputing
Applications at Illinois.

The low-cost, portable, smartphone-integrated system provides a promising solution to address the
challenges of infectious disease diagnostics, especially in resource-limited settings or in situations where
a result is needed immediately. The diagnostic tool’s integration with mobile communications
technology allows personalized patient care and facilitates information management for both
healthcare providers and epidemiological surveillance efforts. Importantly, the system achieves
detection limits comparable to those obtained by laboratory-based methods and instruments and does
so in about 30 minutes.

A useful capability for human point-of-care (POC) diagnosis or for a mobile veterinary laboratory is to
simultaneously test for the presence of more than one pathogen with a single test protocol, which
lowers cost, saves time and effort, and allows for a panel of pathogens, which may cause similar
symptoms to be identified.

Infectious diseases remain the world’s top contributors to human death and disability, and with recent
outbreaks of Zika virus infections, there is a keen need for simple, sensitive and easily translatable POC
tests. The Zika virus appeared in the international spotlight in late 2015 as evidence emerged of a
possible link between an epidemic affecting Brazil and increased rates of microcephaly in newborns. Zika
has become a widespread global problem — the World Health Organization (WHO) documented last
year that since June 2016, 60 nations and territories report ongoing mosquito-borne transmission.
Additionally, since the Zika virus infection shares symptoms with other diseases such as Dengue and
Chikungunya, quick and accurate diagnoses are required to differentiate these infections and to
determine the need for aggressive treatment or quarantine.

For the research effort, horses were used as an animal model for respiratory disease in humans and
food animals.
“You can often more easily develop diagnostic tools for human use by coming in to development from
the animal side of things first,” Nash said. “Many diseases show up first in animals — kind of the canary
in the coal mine.”

A key project contributor, Nash commented on the financial impact of infectious disease outbreaks in
horses: “It’s costly to horse owners and trainers and disrupts the business operations of all equine
sports. Consider this — on December 25, 2016, a single horse stabled at the Fair Grounds Race Course in
New Orleans experienced a fever and subsequently developed neurological symptoms. The state
diagnostic lab was 100 miles away and was closed for the Christmas holiday. The end result was an
equine herpesvirus-1 (EHV-1) outbreak that resulted in the quarantine of over 200 horses at the
racetrack and a serious financial loss for horse owners and the racetrack owner, Churchill Downs, Inc.
Imagine the consequences if they ever had to postpone the Kentucky Derby due to a disease outbreak.”

The technology is intended to enable clinicians to rapidly diagnose disease in their office or in the field,
resulting in earlier, more informed patient management decisions while markedly improving the control
of disease outbreaks. An important prerequisite for the widespread adoption of POC tests at the
patient’s side is the availability of detection instruments that are inexpensive, portable and able to share
data wirelessly over the internet.

The system uses a commercial smartphone to acquire and interpret real-time images of an enzymatic
amplification reaction that takes place in a silicon microfluidic chip that generates green fluorescence
and displays a visual readout of the test. The system is composed of an unmodified smartphone and a
portable 3D–printed cradle that supports the optical and electrical components and is connected to the
rear-facing camera of the smartphone.v

The software application operating on the smartphone gathers information about the tests conducted
on the microfluidic card, patient-specific information, and the results from the assays, that are then
communicated to a cloud storage database.

“This project is a game changer,” Nash said. “This is the future of medicine — empowered front-line
healthcare professionals. We can’t stop viruses and bacteria, but we can diagnose more quickly. We
were able to demonstrate the clear benefit to humankind, as well as to animals, during the proposal
phase of the project, and our results have proved our premise. Next I want to go into the field, multiple
sites, multiple geographic locations, and test in real-world situations.”

Fu Sun, U of I graduate student and research assistant, sees this project as fulfillment of one of her
primary career objectives: “I entered graduate school with the hope to make a better world by
developing biomedical devices that can facilitate effective disease prevention, diagnosis or treatment.
This project is in line with my goal since it provides a point-of-care solution for the fast diagnosis of
infectious diseases. Connected to a cloud database through a smartphone, it helps healthcare providers
in the field embrace the era of big data and the Internet of Things.”

The system represents the only platform to date that can multiplex detection of viral and other nucleic
acid targets on a portable POC setup using one droplet of bodily fluid, including whole blood.

For Nash, the experience of working with the University of Illinois team and other project collaborators
was mutually beneficial. “A diverse team was actually created here,” he said. “A wicked smart group of
people! I can’t envision going into a project without engineers now.”
New handheld TRI-analyzer uses smartphone to detect disease

Posted on August 11, 2017 by bcunning@illinois.edu

Researchers at the University of Illinois at Urbana-Champaign have developed technology that enables a
smartphone to perform lab-grade medical diagnostic tests that typically require large, expensive
instruments. Costing only $550, the spectral transmission-reflectance-intensity (TRI)-Analyzer from
Bioengineering and Electrical & Computer Engineering Professor Brian Cunningham’s lab attaches to a
smartphone and analyzes patient blood, urine, or saliva samples as reliably as clinic-based instruments
that cost thousands of dollars.

“Our TRI-Analyzer is like the Swiss Army knife of biosensing,” said Cunningham, the Donald Biggar
Willett Professor of Engineering and director of the Micro + Nanotechnology Lab at Illinois. “It’s capable
of performing the three most common types of tests in medical diagnostics, so in practice, thousands of
already-developed tests could be adapted to it.”

In a recently published paper, Cunningham’s team used the TRI-Analyzer to perform two commercially
available assays — a test to detect a biomarker associated with pre-term birth in pregnant women and
the PKU test for newborns to indirectly detect an enzyme essential for normal growth and development.
Their tests results were comparable to those acquired with clinic-grade spectrometer instrumentation.

“The TRI-Analyzer is more of a portable laboratory than a specialized device,” said Kenny Long, an
MD/PhD student and lead author of the research study.

Among the many diagnostic tests that can be adapted to their point-of-care smartphone format, Long
said, is an enzyme-linked immunosorbent assay (ELISA), which detects and measures a wide variety of
proteins and antibodies in blood and is commonly used for a wide range of health diagnostics tests. The
system is capable of detecting the output of any test that uses a liquid that changes color, or a liquid
that generates light output (such as from fluorescent dyes).

The TRI-Analyzer operates by converting the smartphone camera into a high-performance

spectrometer. Specifically, the analyzer illuminates a sample fluid with the phone’s internal white LED
flash or with an inexpensive external green laser diode. The light from the sample is collected in an
optical fiber and guided through a diffraction grating into the phone’s rear-facing internal camera. These
optical components are all arranged within a 3D-printed plastic cradle.

The TRI-Analyzer can simultaneously measure multiple samples by using a microfluidic cartridge that
slides through an opening in the back of the cradle. This ability to analyze multiple samples quickly and
reliably makes the Analyzer suitable for patients who lack convenient access to a clinic or hospital with
diagnostic test facilities or for patients with urgent health situations requiring rapid results.

“Our analyzer can scan many tests in a sequence by swiping the cartridge past the readout head, in a
similar manner to the way magnetic strip credit cards are swiped,” said Long.

In addition to its applications in health diagnostics, Cunningham said the TRI Analyzer can also be
applied to point-of use applications that include animal health, environmental monitoring, drug testing,
manufacturing quality control, and food safety. The patented technology is available for license.
 Smartphone biosensors Smartphone biosensors

Since their introduction in 1997, “smart” mobile phones with internet connectivity, high-resolution
cameras, touchscreen displays, and powerful CPUs have gained rapid market acceptance. It is estimated
that, of the ~7.5 billion mobile phones that are currently in use, 51% of them can be classified as
smartphones, with an expected rise to ~76% by 2022 (Ericsson Mobility Report, 2017). The rapid
acceptance of smartphones is driven by a combination of falling prices and increasingly sophisticated
features. In addition, there is a growing ecosystem of applications that take advantage of the phone’s
sensors, display, and connection to powerful computing and cloud-based data storage capabilities. The
built-in capabilities of smartphones can be further extended through the addition of accessories that
enable the phone to sense different types of information, with uses in many different point-of-care testing
applications (POCT). Recent examples include attachments that enable smartphones to serve as
stethoscopes, ultrasound probes, microscopes, fluorescent microscopes, and label-free biosensor
detection instruments. POCT is broadly applicable across industries ranging from medical diagnostics,
food safety, food processing quality control, water quality monitoring, animal diagnostics, and pathogen
detection.

Further incorporation of traditionally laboratory-based biosensing into smartphone platforms has much
potential, as it opens the door for testing in situations not currently feasible and by a much broader range
of users. Such developments may help to facilitate the goal of “personalized medicine,” in which home-
based tests may be used to diagnose a medical condition but with a system that automatically
communicates results to a cloud-based monitoring system that alerts the physician when warranted. Low-
cost portable biosensor systems integrated with smartphones also may enable diagnostic technology that
can be translated to resource-poor regions of the world for pathogen detection, disease diagnosis, and
monitoring of nutritional status. Such a system, deployed widely, would be capable of rapidly monitoring
for the presence of environmental contaminants over large areas, or tracking the development of a
medical condition throughout a large population. Of all the label-free detection approaches that have
been demonstrated, those based upon optical phenomena have been most commercially accepted due
to a combination of sensitivity, sensor cost, detection system robustness, and high throughput.
Adsorption of biomolecules, viral particles, bacteria, or cells on the surface of an optical biosensor
transducer results in a shift in the conditions of optimal optical coupling, which can be measured by
illuminating the transducer surface and subsequently measuring a property of the reflected or transmitted
light. Such a detection approach is extremely robust and has become economically advantageous due to
the advent of low-cost light-emitting diodes, semiconductor lasers, and miniature spectrometers. For
example, surface plasmon resonance (SPR)-based and photonic crystal (PC) optical biosensors are capable
of detecting broad classes of biological analytes through their intrinsic dielectric permittivity. Each
approach has been successfully implemented in the form of large laboratory instruments and miniaturized
(shoebox-sized) systems. Our lab seeks to take such optical sensors and shrink them to handheld
detachments, with our initial successes being in both the PC-based label-free sensing and Enzyme Linked
Immunosorbent Assays (ELISA).
Figure 1. A student uses the smartphone application to compare wavelength shifts in two samples
(foreground, left), measured with a PC-based biosensor (foreground, right). Representative spectra, as
observed by the smartphone showing changes in reflected bands, is indicative of biological absorption
onto a PC surface (background).

Our developed technology uses a cradle-based attachment to allow the onboard camera of a smartphone
to function as a spectrometer. The dispersion of light resulting from transmission through a diffraction
grating allows transmitted light to be spatially differentiated along one dimension of the CMOS sensor.
Changes in wavelength resultant from biological absorption to a PC surface, analogous to the PC-based
bench top apparatuses used throughout the Nanosensors Group, can be measured with minimal
modification to methodologies. Proof of concept has been demonstrated with both regular layers of
polymer deposited on PCs as well as a basic biological absorption of Protein A and porcine IgG antibodies.

Similarly, ELISA procedures can be modified to be read on the smartphone system. Since its introduction,
it has become one of the most widely adaptable tools for biological assays, allowing for the rapid
quantification of proteins and antibodies for diseases ranging from HIV to cancer, yielding over 40,000
new articles involving the technology annually. An ELISA test is completed by immobilizing antibodies that
possess an affinity for a specific biomolecule of interest onto a standard format 96-well microtiter plate
and then passing over a series of liquids, exploiting the high specificity of the antibody-antigen interaction
to eventually yield a colorometric change of the liquid sample based upon the cleavage of a chromogen
moiety by an enzyme. When the optical absorptions of standards at known concentrations are used to
obtain a calibration curve, the concentrations of an analyte within a test sample may be accurately
determined via interpolation. Similarly, other color-specific absorption tests can be modified and read on
the system. So far, we have demonstrated the success of such tests using both IL-6, a useful cancer
biomarker, and Ara h 1, a biomarker responsible for peanut allergies, at physiologically relevant
concentrations.
Figure 2. Steps of an ELISA Procedure. Primary antibodies are attached to the bottom of a 96-well
microtiter plate. Next, molecules of interest are incubated on the sensing surface and attach to exposed
antibodies. Secondary antibodies bound to enzymes capable of catalysing chromogenic reactions are
incubated and attach specifically to the immobilized molecules of interest. Finally, chromogenic
compounds are exposed to the sensor and are catalysed by any bound enzymes generating a color change
that can then be quantified using the smartphone system.

The smartphone biosensor is also capable of conducting fluorescence applications. Light emitters operate
via a variety of optical mechanisms that include fluorescence, chemoluminescence, and semiconductor
quantum dot-excited electron relaxation. Among these approaches, those that utilize the Förster
Resonance Energy Transfer (FRET) as a mechanism for observing changes in the quenching efficiency
between matched donor-acceptor pairs are effective methods for diagnostics applications, because
single-step assays are performed in liquid, without complex mixing-washing steps. For FRET assays, the
ability to measure the spectrum of fluorescent emission is especially useful, as the combined contributions
of donor and acceptor fluorophores can be measured independently, while wavelength-selective filters
are not necessary. By using the function of the smartphone fluorimeter, which can disperse and analyze
the incident optical signal, a sensitive molecular-beacon FRET assay for a specific microRNA sequence can
be performed. Our results show that smartphone-based spectroscopic fluorimetry is a route towards
portable biomolecular assays for viral/bacterial pathogens and toxins. The resulting capability may find
applications that include point-of-care detection/analysis of pathogens, specific nanoparticle detection,
human/animal diagnostics, and food safety.

Figure 3. (a) Spectral bands on the smartphone screen before the target miR-21 is introduced (top;
target miRNA (T): 0 μM), and after addition of the target that is a complementary match to the probe
(bottom; T: 1 μM). (b) Normalized fluorescence intensity distributions. Inset shows magnified spectra
that corresponds to the dashed area. (c) Normalized total intensities according to various T-miRNA
concentrations. The intensity over the wavelength range of 560-580 nm was integrated to derive a
single intensity value for each concentration. Inset shows enlarged region of the area encompassed by
the dashed box.