Beng Koon Ng

In-fiber common-path optical coherence tomography using a conical-tip fiber

Common-path optical coherence tomography (CPOCT) is known to reduce group velocity dispersion and polarization mismatch between the reference and the sample arm as both arms share the same physical path. Existing implementations of CPOCT typically require one to incorporate an additional cover glass within the beam path of the sample arm to provide a reference signal. In this paper, we aim to further reduce this step by directly making use of the back-reflected signal, arising from a conical lens-tip fiber, as a reference signal. The conical lens, which is directly manufactured onto the optical fiber tip via a simple selective-chemical etching process, fulfils two functions acting as both the imaging lens and the self-aligning reference plane. We use a Fourier-domain OCT system to demonstrate the feasibility of this technique upon biological tissue. An in-fiber CPOCT technique may prove potentially useful in endoscopic OCT imaging.

Near-field focusing properties of zone plates in visible regime - New insights

Near-field focusing properties of zone plates are investigated in the visible regime by a 3-dimensional finite-difference time-domain method. It is shown that Frensel zone plates (FZPs) with metallic coatings can achieve subwavelength focusing in the visible wavelength. The characteristics of subwavelength focusing are found to be independent of the type of metal coatings used. All the FZPs exhibit similar shift in focal length and depth of focus when compared with classical calculations. These results indicate that plasmonic waves do not contribute to subwavelength focusing. Instead the subwavelength focusing characteristic is attributed to the interference of diffracted evanescent waves from a large numerical aperture. It is found that the near-field focusing of FZPs suppresses higher order foci such that the corresponding diffraction efficiency is improved. The use of phase zone plate structured on glass without opaque coating is proposed to improve the diffraction efficiency of subwavelength focusing.

Concentration dependence of gold nanoshells on the enhancement of optical coherence tomography images: a quantitative study

The effective use of gold nanoshells as a contrast agent for optical coherence tomography (OCT) may be hampered by the delivery of a wrong dose to tissue that results in unwanted signal attenuation. In this study we examine how changes in mu(s) due to concentration variations affect the OCT image and then define a dosing range that would result in appropriate scattering coefficient, mu(s), to maintain an acceptable signal attenuation level. Our results show that an increase in sample mu(s) not only enhances the OCT signal near the surface but also attenuates the signal deeper into the sample. We synthesized gold nanoshells with an 81 nm radius silica core and 23 nm shell thickness and found that a concentration range of 5.6 x 10(9)<c<2.3 x 10(10) particles/ml provided adequate signal enhancement near the surface without severely compromising the imaging depth due to signal attenuation. We also demonstrate the extraction of mu(s) from the OCT signal to estimate the gold nanoshells concentration in vivo and verified that the estimated concentration of 6.2 x 10(9) particles/ml in a mouse tumor after intravenous delivery lies within this concentration range to effectively enhance the tumor image.

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.

Quantitative diagnosis of cervical neoplasia using fluorescence lifetime imaging on haematoxylin and eosin stained tissue sections.

The use of conventional fluorescence microscopy for characterizing tissue pathological states is limited by overlapping spectra and the dependence on excitation power and fluorophore concentration. Fluorescence lifetime imaging microscopy (FLIM) can overcome these limitations due to its insensitivity to fluorophore concentration, excitation power and spectral similarity. This study investigates the diagnosis of early cervical cancer using FLIM and a neural network extreme learning machine classifier. A concurrently high sensitivity and specificity of 92.8% and 80.2%, respectively, were achieved. The results suggest that the proposed technique can be used to supplement the traditional histopathological examination of early cervical cancer.

Fluorescence lifetime discrimination using expectation-maximization algorithm with joint deconvolution

The fluorescence lifetime technique offers an effective way to resolve fluorescent components with overlapping emission spectra. The presence of multiple fluorescent components in biological compounds can hamper their discrimination. The conventional method based on the nonlinear least-squares technique is unable to consistently determine the correct number of fluorescent components in a fluorescence decay profile. This can limit the applications of the fluorescence lifetime technique in biological assays and diagnoses where more than one fluorescent component is typically encountered. We describe the use of an expectation-maximization (EM) method with joint deconvolution to estimate the fluorescence decay parameters, and the Bayesian information criterion (BIC) to accurately determine the number of fluorescent components. A comprehensive simulation and experimental study is carried out to compare the performance and accuracy of the proposed method. The results show that the EM-BIC method is able to accurately identify the correct number of fluorescent components in samples with weakly fluorescing components.

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.

Photothermal imaging of nanoparticles beyond the diffraction limit

In this paper, a full-field photothermal imaging technique, which does not require a time consuming scan as used in the conventional photothermal imaging system, is reported. Imaging on gold nanoparticles (70 nm) and a blue polystyrene bead (193 nm) were conducted and the experimental results demonstrate the visualization ability of the photothermal imaging technique on nanotargets that are below the diffraction limit. The photothermal imaging system can be operated in an ambient environment where vacuum is not required.

Enhancement of Early Cervical Cancer Diagnosis with Epithelial Layer Analysis of Fluorescence Lifetime Images

This work reports the use of layer analysis to aid the fluorescence lifetime diagnosis of cervical intraepithelial neoplasia (CIN) from H&E stained cervical tissue sections. The mean and standard deviation of lifetimes in single region of interest (ROI) of cervical epithelium were previously shown to correlate to the gold standard histopathological classification of early cervical cancer. These previously defined single ROIs were evenly divided into layers for analysis. A 10-layer model revealed a steady increase in fluorescence lifetime from the inner to the outer epithelial layers of healthy tissue sections, suggesting a close association with cellular maturity. The shorter lifetime and minimal lifetime increase towards the epithelial surface of CIN-affected regions are in good agreement with the absence of cellular maturation in CIN. Mean layer lifetimes in the top-half cervical epithelium were used as feature vectors for extreme learning machine (ELM) classifier discriminations. It was found that the proposed layer analysis technique greatly improves the sensitivity and specificity to 94.6% and 84.3%, respectively, which can better supplement the traditional gold standard cervical histopathological examinations.

Resolving interparticle position and optical forces along the axial direction using optical coherence gating

This research is supported by grants nSCS-BU0052 and RP C-015/2007 from Agency for Science, nTechnology and Research and grant RGM39/06 from nthe Ministry of Education, Singapore.