These technologies all take the form of a separate device from which the optical biosensor draws sample. There is a continuing need for more efficient, small, automated concentrators for targets in a wide variety of sample matrices that can be effectively integrated with the biosensor. Why not integrate the biosensor into the sampler?
The use of microfluidics for sample processing can eliminate the manual operations, reducing variability and expediting the assay as discussed previously. Design software to create microfluidic systems, as opposed to characterizing existing systems, is just beginning to appear. Finding or fabricating reliable pumps that provide the right amount of fluid at an exact and steady flow rate is extremely difficult.
Many of the on-chip pumps and valves reported in the literature are appropriate only for use with relatively clean, well-defined fluids. Pumps and valves can also consume the largest portion of the power required by a system. When the biosensor must run on batteries, energy consumption is a significant issue. Both reducing energy requirements and improving miniature energy sources, such as a microfluidic fuel cell or a biomimetic photocenter, 94 , 95 will be critical for long-term acceptance and practicality of automated biosensor systems.
In addition to enabling automation, the microfluidic components can be integrated with the optical components. Capillary walls can constrain the flow of sample and reagents, provide a surface for the attachment of biorecognition molecules, and serve as waveguides for excitation of bound fluorophores and collection of fluorescence emission. Photopatterning of glass has resulted in the fabrication of both microchannels and optical waveguides in a single substrate.
To date miniaturization and integration of the fluidics and optics has depended on the development of microfluidic fabrication techniques and the miniaturization of optical components such as CCD and CMOS detectors, avalanche photodiodes, and light emitting diodes. While integration of miniaturized silicon devices into biosensors has begun, there is an entire generation of new polymer optical components that will be even more suited to integration with microfluidics and sensor substrates: A biosensor chip for blood typing.
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The blood type is read by eye. Printed with permission from The Royal Society of Chemistry. As optical biosensor systems become smaller and more automated, new capabilities can be achieved by combining two or more analytical technologies. For example, on-chip DNA amplification has been combined with fluorescence detection following capillary electrophoresis of the products. Photonic crystal spectrometers might, for instance, be combined with on-chip chromatography to look at reaction rates for ligand-receptor or protein-protein interactions as part of a general trend to develop systems providing both complex biochemical and optical information.
Today and Tomorrow
Optical biosensors have proven advantages over other types of sensors for multi-target sensing and continuous monitoring. These advantages have highlighted the potential of optical biosensors to address the analytical needs for medical and veterinary diagnostics, food processing, environmental protection and homeland security. Increased awareness of the capabilities of optical biosensors for onsite, multi-analyte sensing is encouraging continued public investment as well as attracting larger amounts of private development funds.
The largest initial market for commercial optical biosensors was the research community. Rich and Myszka documented the number of papers published over the six years beginning in that used commercial optical biosensors. Already, smaller, less expensive, and more user-friendly systems are appearing particularly for nucleic acid amplification and identification, affinity microarray analysis, and flow cytometry.
Medical applications are currently attracting the most commercial interest, particularly in terms of implementing the capacity of optical biosensors for multiplexed diagnostics. However, unless the diagnostic test is appropriate for screening large populations Figure 3 or is tied to a high-value drug, then it is difficult to generate sufficient profit to motivate a company to go through the regulatory process, assume liability, and expend marketing capital.
- THE “BIO” IN THE OPTICAL BIOSENSOR?
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Diagnostic biosensors that simultaneously test for large numbers of infectious diseases or biomarkers face the additional regulatory challenge of demonstrating that the information provided is of clinical significance. In many of these highly multiplexed tests, there are not sufficient patients with each of the diseases included in the panel to get statistically significant information, and comprehensive clinical trials would be prohibitively expensive.
In the case of biomarkers, the etiology of the appearance of these markers in the disease may not yet be well understood or indeed the importance of each marker well validated. Biomarker analysis could be especially valuable in prognosis and risk analysis e. As more and more diagnostic techniques become available for point-of-care and self-testing, , physicians and patients must be educated to use such tools effectively. Already, optical biosensors have been commercially developed to test for infectious disease, alcohol, drugs of abuse, and heart attack; feedback from physicians and patients will pave the way for improved biosensors.
Concept for a biosensor with automated sample processing unit developed by the author and a polymer photodiode array developed by BioIdent. Surprisingly, the goal of using optical biosensors for diagnostics in developing countries has become an important driver for engineering more cost-effective systems Table 1. Successful approaches will work just as well in technologically advanced cultures and be more rapidly accepted due to the lower cost of use.
However, again it must be emphasized that in order for such biosensors to be accepted, the information they provide must be actionable. Laboratory structure constraints in low-resource settings informing product attributes Reprinted with permission from Annual Reviews www. Optical biosensors are also being used to a growing degree in other application areas. Environmental concerns are driving the development of on-site monitoring systems to reduce the response time and cost of pollution control in comparison to shipping samples to a central laboratory.
Optical biosensors are being tested for monitoring air, water, and soil, with the primary interest coming from environmental regulatory agencies. Food testing applications have been clearly demonstrated, but the regulatory agencies responsible for monitoring food safety are generally too underfunded and understaffed to devote significant resources to implementing new technology. Food processing companies are interested in automated monitoring systems to promote safer products and reduce liability, but sampling presents a major challenge. While a few companies have emplaced optical biosensors for food testing, the efficacy of these systems for process monitoring is just beginning to be confirmed.
While the technical future of optical biosensors is in the hands of clever scientists and inventive engineers, the rate of transition to the user community will be controlled by a wide variety of nonscientific factors.
Social concerns over problems such as resource depletion will also drive priorities for system design as well as for application areas. For example, as energy becomes more and more expensive, on-chip power generation becomes more attractive. As clean water becomes more and more of a problem, testing of drinking water will assume a higher priority and more thorough testing will be publicly demanded than is currently the case. Both public and private funding priorities will react accordingly to public concerns. For those of us working to develop optical biosensors, the number of opportunities to incorporate new science and technology into our systems is almost overwhelming.
The only limitations seem to be our ability to integrate basic and cutting-edge information from disciplines other than our own, to find colleagues willing to work with us who have the skills we lack, and to find the financial and physical resources to create and test new optical biosensors. However, we must also consider the ultimate user, the reliability of the data produced, and the impact of any reaction to that data—positive or negative.
Such considerations can focus our research and development efforts into the most productive paths to produce optical biosensors that can solve real problems in everyday life. Jeffrey Erickson and Chris Taitt for careful critique of the manuscript. National Center for Biotechnology Information , U. Author manuscript; available in PMC Jan The publisher's final edited version of this article is available at Anal Chem. See other articles in PMC that cite the published article. Abstract Optical biosensors have begun to move from the laboratory to the point of use.
Open in a separate window. Table 1 Laboratory structure constraints in low-resource settings informing product attributes Reprinted with permission from Annual Reviews www. Laboratory infrastructure constraints in low-resource settings Implications on point-of-care diagnostic product attributes A wide disparity of laboratory facilities and capacities within a country and among countries Careful consideration for the final user of the test is required.
Poor or nonexistent external quality control and laboratory accreditation systems The test should be reproducible and provide clear and easy to interpret internal and process controls. Unreliable procurement system leading to stock outs of key laboratory supplies The test should require as few external reagents and supplies as possible.
Unreliable quality of reagents and supplies procured through national channels The test should require as few external reagents and supplies as possible. Lack of basic essential equipment The test should require as little instrumentation as possible or provide its own instrumentation. Lack of laboratory consumables No assumptions should be made regarding supplies for specimen collection, storage, and handling.
Unreliable water supply and quality This is extremely variable in different regions and seasons, and a device should not require external water if high quality is needed. Unreliable power supply and quality This is often tied to water supply. Devices requiring external power should account for long periods of time without network electricity supply and variability as well as frequency of surges from the network electricity supply. Inconsistent refrigeration capacity This is associated with unreliable power supply.
Insufficiently skilled staff The test should be easy to use and interpret. Limited training opportunities Any training requirements should be given special consideration for the introduction strategy. Poor waste-management facilities The environmental impact of disposable, chemical reagents, and biohazardous materials should be considered. Oxford University Press; Biosensors with Fiber Optics. Fiber Optic Chemical Sensors and Biosensors. Current Opinion in Chemical Biology. Brody EN, Gold L. Reviews in Molecular Biotechnology.
Rajendran M, Ellington AD. Engineering in Medicine and Biology Magazine. Lu J, Rosenzweig Z. Kwon S, Lee LP. Song S, Singh AK. Nguyen NT, Wu Z. Lab on a Chip. Falconi C, Fratini M. Atencia J, Beebe DJ. Xia Y, Whitesides GM. Heckele M, Schomburg WK. International Communications in Heat and Mass Transfer. Erickson D, Li D. Advanced Materials FRG ; This platform was applied in detecting the binding of influenza A virus strains with a panel of glycans of diverse structures. The apparent equilibrium dissociation constants avidity constants, 10— pM were used as characterizing parameters of viral receptor profiles [ 28 ].
Microarray biosensors based on total internal reflection imaging ellipsometry for the detection of the serum tumour biomarker CA had an estimated detection limit of CA of Reflectometric interference spectroscopy RIfS is a label-free and time-resolved method where the simple optical set-up is based on white light interference at thin layers.
Changes in the phase and amplitude of polarized light provides information about the thickness and refractive index of the adsorbed protein layer. This method was used for the detection and quantification of diclofenac in bovine milk and the obtained limit of detection was 0. Surface-enhanced Raman scattering SERS is a biosensing technique which enhances the intensity of the vibration spectra of a molecule by several orders of magnitude when it is in close proximity to nano-roughened metallic surfaces or nanoparticles made of gold or silver.
Optical biosensors are expected to grow most in importance in the healthcare, biomedical and biopharmaceutical sectors. They can provide new analytical tools with reduced size as well as facilitate large-scale high-throughput sensitive screening of a very wide range of samples for many different parameters [ 34 ]. On the other hand, the broad practical application of optical biosensors is still under development and is limited mostly to academic and pharmaceutical environments. A summary of key points of each optical biosensor reviewed here, with examples of use in studying biological problems, is given in Table 1.
When developing new optical biosensor devices for practical applications all the methodological and practical aspects such as robustness, reproducibility, simplicity and shelf life should be carefully taken into account. The main challenge for further research and innovation in the field of optical biosensors is that the optical detection principle allows construction of sensitive, simple and cheap analytical devices with a wide variety of possible applications in portable biosensor systems for in situ screening and monitoring, for example, in personalized medicine, remote areas or in developing countries where the availability of inexpensive diagnostic tools could save many lives.
This publication is the result of the project implementation: The Authors declare that there are no competing interests associated with the manuscript. National Center for Biotechnology Information , U.
Published online Jun This article has been cited by other articles in PMC. Introduction Optical biosensors offer great advantages over conventional analytical techniques because they enable the direct, real-time and label-free detection of many biological and chemical substances. Open in a separate window. Surface plasmon resonance biosensors The physical phenomenon of SPR was first observed in A schematic illustration of the set-up for the SPR imaging reprinted with permission from Elsevier [ 9 ]. Evanescent wave fluorescence biosensors In these biosensors, the biological recognition and the consequent binding event occur within the confines of an evanescent wave.
Schematic diagram of an evanescent wave planar optical waveguide chip reprinted with permission from NPG [ 22 ]. Bioluminescent optical fibre biosensors This technique uses recombinant bioluminescent cells and the bioluminescent signal is transferred from the analyte by an optical fibre. Other optical biosensors Optical waveguide interferometric biosensors An integrated planar optical waveguide interferometric biosensor is a combination of evanescent field sensing and optical phase difference measurement methods.
Ellipsometric biosensors An ellipsometric biosensor measures changes in the polarization of light when it is reflected from a surface. Reflectometric interference spectroscopy biosensors Reflectometric interference spectroscopy RIfS is a label-free and time-resolved method where the simple optical set-up is based on white light interference at thin layers. Surface-enhanced Raman scattering biosensors Surface-enhanced Raman scattering SERS is a biosensing technique which enhances the intensity of the vibration spectra of a molecule by several orders of magnitude when it is in close proximity to nano-roughened metallic surfaces or nanoparticles made of gold or silver.
Conclusions Optical biosensors are expected to grow most in importance in the healthcare, biomedical and biopharmaceutical sectors. Key points of reviewed optical biosensors including selected biological applications. Biosensor Multiplexing Commercialization Label-free? Summary This article includes a brief classification, description and examples of the applications of optical biosensors in medicine, pharmacy, food safety, the environment, biotechnology, defence and security. Surface plasmon resonance biosensors, localized surface plasmon resonance biosensors, evanescent wave fluorescence and bioluminescent optical fibre biosensors, interferometric, ellipsometric, reflectometric interference spectroscopy and surface enhanced Raman scattering biosensors were discussed.
Optical biosensors allow the sensitive and selective detection of a wide range of analytes including viruses, toxins, drugs, antibodies, tumour biomarkers, and tumour cells. Optical biosensors provide new analytical tools with reduced size as well as facilitate large-scale high-throughput sensitive screening of a wide range of samples for many different parameters. The optical detection principle allows construction of simple and cheap analytical devices with many potential applications in portable biosensor systems for in situ detection which can be used in personalized medicine, remote areas and developing countries.
Competing Interests The Authors declare that there are no competing interests associated with the manuscript. Surface plasmon resonance for gas detection and biosensing. Handbook of Surface Plasmon Resonance. Royal Society of Chemistry; Surface plasmon resonance biosensor for the detection of VEGFR-1—a protein marker of myelodysplastic syndromes. A label-free and portable multichannel surface plasmon resonance immunosensor for on site analysis of antibiotics in milk samples. A surface plasmon resonance based biochip for the detection of patulin toxin.
Quantification of arsenic III in aqueous media using a novel hybrid platform comprised of radially porous silica particles and a gold thin film. Application of surface plasmon resonance for the detection of carbohydrates, glycoconjugates, and measurement of the carbohydrate-specific interactions: Dextran hydrogel coated surface plasmon resonance imaging SPRi sensor for sensitive and label-free detection of small molecule drugs. Gold nanorod-based localized surface plasmon resonance biosensors: Localized surface plasmon resonance sensors.
Trends and challenges of refractometric nanoplasmonic biosensors: Localized surface plasmon resonance as a biosensing platform for developing countries. High-resolution biosensor based on localized surface plasmons. Highly sensitive localized surface plasmon resonance immunosensor for label-free detection of HIV Evanescent wave fluorescence biosensors: Rapid multiplexed immunoassay for simultaneous serodiagnosis of HIV-1 and coinfections. Aptamer-based optical biosensor for rapid and sensitive detection of 17beta-estradiol in water samples.
A reusable evanescent wave immunosensor for highly sensitive detection of bisphenol A in water samples. A lower limit of detection for atrazine was obtained using bioluminescent reporter bacteria via a lower incubation temperature. Optical imaging fiber-based live bacterial cell array biosensor. Integrated planar optical waveguide interferometer biosensors: Microfluidic resonant waveguide grating biosensor system for whole cell sensing.
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Detection of avian influenza virus using an interferometric biosensor. Characterization of receptor binding profiles of influenza a viruses using an ellipsometry-based label-free glycan microarray assay platform. Serum tumor marker detection on PEGylated lipid membrane using biosensor based on total internal reflection imaging ellipsometry. Label-free optical biosensor for detection and quantification of the non-steroidal anti-inflammatory drug diclofenac in milk without any sample pretreatment. Label-free reflectometric interference microchip biosensor based on nanoporous alumina for detection of circulating tumour cells.
Kinetic analysis of biointeractions Antigens in clinical samples Proteins in biological samples Xenobiotics and toxins in food Carbohydrate-specific interactions.
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Screening of biomarkers and therapeutic targets Screening of drug—target protein interactions. Clinical diagnostics, biodefence, food testing Clinical biomarkers Toxin screening. Response of cells to genotoxic agents Multidetection of genotoxins by live cell array. Characterizing viral receptor profiles Detection of serum tumour biomarker.