Circulating microRNA breast cancer biomarker detection in patient sera with surface plasmon resonance imaging biosensor.

In this article, we report for the first time, the detection of circulating miRNA as a breast cancer biomarker in patient sera using surface plasmon resonance imaging biosensor. The advantage of this approach lies in the rapid, label-free and sensitive detection. The sensor excites plasmonic resonance on the gold sensor surface and specific DNA-miRNA molecular bindings elucidate responses in the plasmonic resonance image. Experiments of detecting synthetic miRNA molecules (miR-1249) were performed and the sensor resolution was found to be 63.5 nM. The sensor was further applied to screen 17 patient serum samples from National Cancer Centre Singapore and Tan Tock Seng Hospital. Sensor intensity response was found to differ by 20% between malignant and benign cases and thus forms, a potential and an important metric in distinguishing benignity and malignancy.

A Phase-Intensity Surface Plasmon Resonance Biosensor for Avian Influenza A (H5N1) Detection.

In this paper, we present a phase-intensity surface plasmon resonance (SPR) biosensor and demonstrate its use for avian influenza A (H5N1) antibody biomarker detection. The sensor probes the intensity variation produced by the steep phase response at surface plasmon excitation. The prism sensor head is fixed between a pair of polarizers with a perpendicular orientation angle and a forbidden transmission path. At SPR, a steep phase change is introduced between the p- and s-polarized light, and this rotates the polarization ellipse of the transmission beam. This allows the light at resonance to be transmitted and a corresponding intensity change to be detected. Neither time-consuming interference fringe analysis nor a phase extraction process is required. In refractive index sensing experiments, the sensor resolution was determined to be 6.3 × 10-6 refractive index values (RIU). The sensor has been further applied for H5N1 antibody biomarker detection, and the sensor resolution was determined to be 193.3 ng mL-1, compared to 1 µg mL-1 and 0.5 µg mL-1, as reported in literature for influenza antibody detection using commercial Biacore systems. It represents a 517.3% and 258.7% improvement in detection limit, respectively. With the unique features of label-free, real-time, and sensitive detection, the phase-intensity SPR biosensor has promising potential applications in influenza detection.

Diffuse Optical Spectroscopy and Imaging to Detect and Quantify Adipose Tissue Browning.

Adipose (fat) tissue is a complex metabolic organ that is highly active and essential. In contrast to white adipose tissue (WAT), brown adipose tissue (BAT) is deemed metabolically beneficial because of its ability to burn calories through heat production. The conversion of WAT-resident adipocytes to "beige" or "brown-like" adipocytes has recently attracted attention. However, it typically takes a few days to analyze and confirm this browning of WAT through conventional molecular, biochemical, or histological methods. Moreover, accurate quantification of the overall browning process is not possible by any of these methods. In this context, we report the novel application of diffuse reflectance spectroscopy (DRS) and multispectral imaging (MSI) to detect and quantify the browning process in mice. We successfully demonstrated the time-dependent increase in browning of WAT, following its induction through ß-adrenergic agonist injections. The results from these optical techniques were confirmed with those of standard molecular and biochemical assays, which measure gene and protein expression levels of UCP1 and PGC-1α, as well as with histological examinations. We envision that the reported optical methods can be developed into a fast, real time, cost effective and easy to implement imaging approach for quantification of the browning process in adipose tissue.

Molecular photoacoustic imaging of breast cancer using an actively targeted conjugated polymer.

Conjugated polymers (CPs) are upcoming optical contrast agents in view of their unique optical properties and versatile synthetic chemistry. Biofunctionalization of these polymer-based nanoparticles enables molecular imaging of biological processes. In this work, we propose the concept of using a biofunctionalized CP for noninvasive photoacoustic (PA) molecular imaging of breast cancer. In particular, after verifying the PA activity of a CP nanoparticle (CP dots) in phantoms and the targeting efficacy of a folate-functionalized version of the same (folate-CP dots) in vitro, we systemically administered the probe into a folate receptor-positive (FR+ve) MCF-7 breast cancer xenograft model to demonstrate the possible application of folate-CP dots for imaging FR+ve breast cancers in comparison to CP dots with no folate moieties. We observed a strong PA signal at the tumor site of folate-CP dots-administered mice as early as 1 hour after administration as a result of the active targeting of the folate-CP dots to the FR+ve tumor cells but a weak PA signal at the tumor site of CP-dots-administered mice as a result of the passive accumulation of the probe by enhanced permeability and retention effect. We also observed that folate-CP dots produced ~4-fold enhancement in the PA signal in the tumor, when compared to CP dots. These observations demonstrate the great potential of this active-targeting CP to be used as a contrast agent for molecular PA diagnostic imaging in various biomedical applications.

Non-labeling multiplex surface enhanced Raman scattering (SERS) detection of volatile organic compounds (VOCs).

In this paper, we report multiplex SERS based VOCs detection with a leaning nano-pillar substrate. The VOCs analyte molecules adsorbed at the tips of the nano-pillars produced SERS signal due to the field enhancement occurring at the localized surface plasmon hot spots between adjacent leaning nano-pillars. In this experiment, detections of acetone and ethanol vapor at different concentrations were demonstrated. The detection limits were found to be 0.0017 ng and 0.0037 ng for ethanol and acetone vapor molecules respectively. Our approach is a non-labeling method such that it does not require the incorporation of any chemical sensing layer for the enrichment of gas molecules on sensor surface. The leaning nano-pillar substrate also showed highly reproducible SERS signal in cyclic VOCs detection, which can reduce the detection cost in practical applications. Further, multiplex SERS detection on different combination of acetone and ethanol vapor was also successfully demonstrated. The vibrational fingerprints of molecular structures provide specific Raman peaks for different VOCs contents. To the best of our knowledge, this is the first multiplex VOCs detection using SERS. We believe that this work may lead to a portable device for multiplex, specific and highly sensitive detection of complex VOCs samples that can find potential applications in exhaled breath analysis, hazardous gas analysis, homeland security and environmental monitoring.

Multifunctional photosensitizer-based contrast agents for photoacoustic imaging.

Photoacoustic imaging is a novel hybrid imaging modality combining the high spatial resolution of optical imaging with the high penetration depth of ultrasound imaging. Here, for the first time, we evaluate the efficacy of various photosensitizers that are widely used as photodynamic therapeutic (PDT) agents as photoacoustic contrast agents. Photoacoustic imaging of photosensitizers exhibits advantages over fluorescence imaging, which is prone to photobleaching and autofluorescence interference. In this work, we examined the photoacoustic activity of 5 photosensitizers: zinc phthalocyanine, protoporphyrin IX, 2,4-bis [4-(N,N-dibenzylamino)-2,6-dihydroxyphenyl] squaraine, chlorin e6 and methylene blue in phantoms, among which zinc phthalocyanine showed the highest photoacoustic activity. Subsequently, we evaluated its tumor localization efficiency and biodistribution at multiple time points in a murine model using photoacoustic imaging. We observed that the probe localized at the tumor within 10 minutes post injection, reaching peak accumulation around 1 hour and was cleared within 24 hours, thus, demonstrating the potential of photosensitizers as photoacoustic imaging contrast agents in vivo. This means that the known advantages of photosensitizers such as preferential tumor uptake and PDT efficacy can be combined with photoacoustic imaging capabilities to achieve longitudinal monitoring of cancer progression and therapy in vivo.

Colorimetric surface plasmon resonance imaging (SPRI) biosensor array based on polarization orientation.

A colorimetric surface plasmon resonance (SPR) imaging biosensor array based on polarization orientation rotation is presented in this paper. It measures the spectral characteristic variations caused by the steep phase difference between the p- and s-polarization occurring at surface plasmon excitation. It provides one-order of magnitude sensor resolution improvement comparing to existing phase-sensitive SPR imaging sensors and the two-dimensional (2D) sensing capability of the imaging sensor enables multiplex, high throughput array based simultaneous detection for a range of different bio-molecular interactions. Experiments on the binding interactions detection between anti-bovine serum albumin (anti-BSA) and BSA antigen have been performed. All binding interactions occurred at 5×4 protein array were real-time monitored simultaneously. A sensor resolution of 8.26ng/ml (125pM) has been demonstrated, which is one-order of magnitude (12 times) better than the detection limit reported by existing phase-sensitive SPR imaging sensors in the literature, while no time-consuming phase modulation and phase extraction processes are required. Furthermore, the optical colorimetric image read-out of the sensor is easy to be identified by the end users comparing to conventional intensity or phase information. The colorimetric SPR imaging biosensor array can find promising potential applications in high throughput clinical disease diagnosis, protein biomarkers screening and drug screening.

Multiplex spectral surface plasmon resonance imaging (SPRI) sensor based on the polarization control scheme.

A two-dimensional (2D) spectral SPR sensor based on a polarization control scheme is reported in this paper. The polarization control configuration converts the phase difference between p- and s- polarization occurring at surface plasmon resonance (SPR) into corresponding color responses in spectral SPR images. A sensor resolution of 2.7 x 10(-6) RIU has been demonstrated, which corresponds to more than one order of magnitude resolution improvement (26 times) comparing to existing 2D spectral SPR sensors. Multiplex array detection has also been demonstrated with the spectral SPR imaging sensor. In a 8 x 4 sensor array, 32 samples with different refractive index values were monitored simultaneously. Detection on bovine serum albumin (BSA) antigen-antibody binding further demonstrated the multiplex detection capability of the 2D spectral SPR sensor for bio-molecular interactions. The detection limit is found to be 21 ng/ml, which is 36 times better than the detection limit previously reported by phase imaging SPR sensors. In light of the advantages of high sensitivity, 2D multiplex detection and real-time response, the spectral SPR imaging sensor can find promising applications in rapid, high throughput, non-labeling and multiplex detection of protein array for proteomics studies, biomarker screening, disease prognosis, and drug discovery.

Application of surface plasmon resonance sensing to studying elastohydrodynamic lubricant films.

We present a new technique based on the spectral characteristics associated with the surface plasmon resonance (SPR) effect for studying lubricants in elastohydrodynamic (EHD) dimples. The pressure inside the EHD dimple causes a localized change of the refractive index (RI) of the entrapped lubricant. This also results in a shift in the spectral SPR absorption dip. By monitoring the color changes within the SPR image, one can obtain a direct measurement of the RI of the entrapped lubricant, from which the pressure distribution within the elastohydrodynamic lubrication (EHL) dimple can be deduced by means of a predetermined relation of pressure and RI of the tested lubricant. Dimples formed with the lubricants PB 2400 and H 1900 were studied in our experiments. Because SPR is sensitive only to the RI variation within a thin region (approximately one wavelength) close to the sensor's surface, the new technique does not require any measurement of the absolute film thickness of the lubricant. This is much simpler than the existing two-beam interferometric technique for measuring the RI of lubricants in EHD dimples, which requires simultaneous measurements of optical film thickness by use of two beams of different angles of incidence. In light of this advantage we anticipate that the new technique can be applied to pressure field mapping in highly loaded rolling and sliding EHL contacts.