The Application of an Extracellular Vesicle-Based Biosensor in Early Diagnosis and Prediction of Chemoresponsiveness in Ovarian Cancer.

BACKGROUND: Ovarian cancer (OVCA) is the most fatal gynecological cancer with late diagnosis and plasma gelsolin (pGSN)-mediated chemoresistance representing the main obstacles to treatment success. Since there is no reliable approach to diagnosing patients at an early stage as well as predicting chemoresponsiveness, there is an urgent need to develop a diagnostic platform for such purposes. Small extracellular vesicles (sEVs) are attractive biomarkers given their potential accuracy for targeting tumor sites. METHODS: We have developed a novel biosensor which utilizes cysteine-functionalized gold nanoparticles that simultaneously bind to cisplatin (CDDP) and plasma/cell-derived EVs, affording us the advantage of predicting OVCA chemoresponsiveness, and early diagnosis using surface-enhanced Raman spectroscopy. RESULTS: We found that pGSN regulates cortactin (CTTN) content resulting in the formation of nuclear- and cytoplasmic-dense granules facilitating the secretion of sEVs carrying CDDP; a strategy used by resistant cells to survive CDDP action. The clinical utility of the biosensor was tested and subsequently revealed that the sEV/CA125 ratio outperformed CA125 and sEV individually in predicting early stage, chemoresistance, residual disease, tumor recurrence, and patient survival. CONCLUSION: These findings highlight pGSN as a potential therapeutic target and provide a potential diagnostic platform to detect OVCA earlier and predict chemoresistance; an intervention that will positively impact patient-survival outcomes.

Rapid detection of bacteria using gold nanoparticles in SERS with three different capping agents: Thioglucose, polyvinylpyrrolidone, and citrate.

The increase in outbreaks of emerging and re-emerging bacterial infections over the last few decades calls for their rapid detection and treatment. Surface-enhanced Raman spectroscopy (SERS) is a technique that can be applied to develop fast screening systems for bacterial presence in biological samples. Optimizing the capping agents in nanoparticle synthesis is important because capping agents are responsible for controlling the morphological features and chemical properties of the nanoparticles that are essential for SERS. To the best of our knowledge, this paper is the first to study the application of gold nanoparticles capped with thioglucose and polyvinylpyrrolidone (PVP) in SERS detection of bacteria as an alternative to the citrate-capped gold nanoparticles that are often used in SERS detection of bacteria. Three different species of bacteria were used in this study: Cutibacterium acnes, Escherichia coli and Staphylococcus aureus (methicillin-sensitive and methicillin-resistant). This study demonstrates that the thioglucose, citrate both show good contribution in bacterial species identification and the thioglucose shows the best among the three capping agents in two types of S. aureus identification. Moreover, although PVP showed high Raman peaks in the SERS spectrum for each type of bacteria, it showed least contribution in identifying species and strains due to its low efficacy in producing responses from different nucleic acid components in the bacteria cells.

Surface-enhanced Raman scattering for the detection of polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is a multi-factorial heterogeneous syndrome that affects many women of reproductive age. This work demonstrates how the surface-enhanced Raman scattering (SERS) technique can be used to differentiate between PCOS and non-PCOS patients. We determine that the use of SERS, in conjunction with partial least squares (PLS) and principal component analysis (PCA), allows us to detect PCOS in patient samples. Although the role of chemerin in the pathogenesis of PCOS patients is not clear, this work enables us to measure their chemerin levels using the PLS regression method.

An improved partial least-squares regression method for Raman spectroscopy.

It is known that the performance of partial least-squares (PLS) regression analysis can be improved using the backward variable selection method (BVSPLS). In this paper, we further improve the BVSPLS based on a novel selection mechanism. The proposed method is based on sorting the weighted regression coefficients, and then the importance of each variable of the sorted list is evaluated using root mean square errors of prediction (RMSEP) criterion in each iteration step. Our Improved BVSPLS (IBVSPLS) method has been applied to leukemia and heparin data sets and led to an improvement in limit of detection of Raman biosensing ranged from 10% to 43% compared to PLS. Our IBVSPLS was also compared to the jack-knifing (simpler) and Genetic Algorithm (more complex) methods. Our method was consistently better than the jack-knifing method and showed either a similar or a better performance compared to the genetic algorithm.

Effect of narrow spectral filter position on the characteristics of active similariton mode-locked femtosecond fiber laser.

A significant change in active similariton characteristics, both numerically and experimentally, is observed as a function of the location of the lumped spectral filter. The closer the spectral filter is to the input of the Yb(3+)-doped fiber, the shorter the de-chirped pulse width. The peak power of the de-chirped pulse has its maximum value at a certain location of the spectral filter. Four different positions of the spectral filter inside the laser cavity have been theoretically studied and two of them have been verified experimentally.

Hollow core photonic crystal fiber for monitoring leukemia cells using surface enhanced Raman scattering (SERS).

The present paper demonstrates an antibody-free, robust, fast, and portable platform for detection of leukemia cells using Raman spectroscopy with a 785-nm laser diode coupled to a hollow core photonic crystal (HC-PCF) containing silver nanoparticles. Acute myeloid leukemia is one of the most common bone marrow cancers in children and youths. Clinical studies suggest that early diagnosis and remission evaluation of myoblasts in the bone marrow are pivotal for improving patient survival. However, the current protocols for leukemic cells detection involve the use of expensive antibodies and flow cytometers. Thus, we have developed a new technology for detection of leukemia cells up to 300 cells/ml using a compact fiber HC-PCF, which offers a novel alternative to existing clinical standards. Furthermore, we were also able to accurately distinguish live, apoptotic and necrotic leukemic cells.

Determining optimum operating conditions of the polarization-maintaining fiber with two far-lying zero dispersion wavelengths for CARS microscopy.

Single femtosecond laser-based coherent anti-Stokes Raman scattering (CARS) microscopy, using a photonic crystal fiber (PCF) pumped in the near-IR to generate a supercontinuum for the Stokes source, is rapidly being adopted as a cost-effective approach. A PCF with two closely-lying zero dispersion wavelengths is a popular choice for the Stokes source, but it is often limited to imaging lipids. A polarization-maintaining PCF with two far-lying zero dispersion wavelengths offers important advantages for polarization CARS microscopy, and for CARS imaging in the fingerprint region. This PCF fiber, though commercially available, has limited use for CARS microscopy in the C-H bond region. The main problem is that the supercontinuum from this fiber is typically noisier than that from a standard PCF with two closely-lying zero dispersion wavelengths. To overcome this, we determined the optimum operating conditions for generating a low-noise supercontinuum out of a PCF with two far-lying zero dispersion wavelengths, in terms of the input parameters of the excitation pulse. We measured the relative intensity noise (RIN) of the Stokes and the corresponding CARS signal as a function of the input laser parameters in this fiber. We showed that the results of CARS imaging using this alternate fiber are comparable to those achieved using the standard fiber, for input laser pulse conditions of low average power, narrow pulse width with slightly positive chirp, and polarization direction parallel to the slow axis of the selected fiber.

Portable, miniaturized, fibre delivered, multimodal CARS exoscope.

We demonstrate for the first time, a portable multimodal coherent anti-Stokes Raman scattering microscope (exoscope) for minimally invasive in-vivo imaging of tissues. This device is based around a micro-electromechanical system scanning mirror and miniaturized optics with light delivery accomplished by a photonic crystal fibre. A single Ti:sapphire femtosecond pulsed laser is used as the light source to produce CARS, two photon excitation fluorescence and second harmonic generation images. The high resolution and distortion-free images obtained from various resolution and bio-samples, particularly in backward direction (epi) successfully demonstrate proof of concept, and pave the path towards future non or minimally-invasive in vivo imaging.

Hollow core photonic crystal fiber as a reusable Raman biosensor.

We report that a single hollow core photonic crystal fiber (HC-PCF) can be used for repetitive characterization of multiple samples by Raman spectroscopy. This was achieved by integrating the HC-PCF to a differential pressure system that allowed effective filling, draining and re-filling of samples into a HC-PCF under identical optical conditions. Consequently, high-quality and reliable spectral data could be obtained which were suitable for multivariate analysis (partial least squares). With the present scheme, we were able to accurately predict different concentrations of heparin and adenosine in serum. Thus the detection scheme as presented here paves a path for the inclusion of HC-PCFs in point-of-care technologies and environmental monitoring where rapid sample characterization is of utmost importance.

Raman spectroscopy for clinical-level detection of heparin in serum by partial least-squares analysis.

Heparin is the most widely used anti-coagulant for the prevention of blood clots in patients undergoing certain types of surgeries including open heart surgeries and dialysis. The precise monitoring of heparin amount in patients' blood is crucial for reducing the morbidity and mortality in surgical environments. Based upon these considerations, we have used Raman spectroscopy in conjunction with partial least squares (PLS) analysis to measure heparin concentration at clinical level which is less than 10 United States Pharmacopeia (USP) in serum. The PLS calibration model was constructed from the Raman spectra of different concentrations of heparin in serum. It showed a high coefficient of determination (R2>0.91) between the spectral data and heparin level in serum along with a low root mean square error of prediction ~4 USP/ml. It enabled the detection of extremely low concentrations of heparin in serum (~8 USP/ml) as desirable in clinical environment. The proposed optical method has the potential of being implemented as the point-of-care testing procedure during surgeries, where the interest is to rapidly monitor low concentrations of heparin in patient's blood.

Monitoring of heparin concentration in serum by Raman spectroscopy within hollow core photonic crystal fiber.

The feasibility of using hollow core photonic crystal fiber (HC-PCF) in conjunction with Raman spectroscopy has been explored for real time monitoring of heparin concentration in serum. Heparin is an important blood anti-coagulant whose precise monitoring and controlling in patients undergoing cardiac surgery and dialysis is of utmost importance. Our method of heparin monitoring offers a novel alternative to existing clinical procedures in terms of accuracy, response time and sample volume. The optical design configuration simply involves a 785-nm laser diode whose light is coupled into HC-PCF filled with heparin-serum mixtures. By non-selectively filling HC-PCF, a strong modal field overlap is obtained. Consequently, an enhanced Raman signal (>90 times) is obtained from various heparin-serum mixtures filled HC-PCFs compared to its bulk counterpart (cuvette). The present scheme has the potential to serve as a 'generic biosensing tool' for diagnosing a wide range of biological samples.

Wavelength remodulation scheme using DPSK downstream and upstream for DWDM-PONs.

We propose a novel wavelength-division-multiplexed passive optical network (WDM-PON) architecture with enhanced tolerance toward chromatic dispersion where a DPSK-modulated downstream signal with constant intensity is remodulated at the ONU side with a return to zero (RZ-DPSK). Driving the downstream modulator with a 50% RZ data enabled us to employ the pulse carver at the ONU for both removing downstream data and generating the optical RZ signal for upstream. This offers an attractive alternative to earlier proposed schemes as it allows us to use full modulation depth (FMD) and balanced detection for downstream data restoration. We experimentally demonstrate the system with both balanced and single-ended detection at 2.5 Gb/s. Error-free operation has been achieved along a 20 Km single mode fiber without dispersion compensation.

Miniaturized multimodal CARS microscope based on MEMS scanning and a single laser source.

We demonstrate a novel miniaturized multimodal coherent anti-Stokes Raman scattering (CARS) microscope based on microelectromechanical systems (MEMS) scanning mirrors and custom miniature optics. A single Ti:sapphire femtosecond pulsed laser is used as the light source to produce the CARS, two photon excitation fluorescence (TPEF) and second harmonic generation (SHG) images using this miniaturized microscope. The high resolution and distortion-free images obtained from various samples such as a USAF target, fluorescent and polystyrene microspheres and biological tissue successfully demonstrate proof of concept, and pave the path towards future integration of parts into a handheld multimodal CARS probe for non- or minimally-invasive in vivo imaging.

Steady and oscillating multiple dissipative solitons in normal-dispersion mode-locked Yb-doped fiber laser.

Multiple dissipative solitons are numerically studied in the normal-cavity-dispersion Yb-doped fiber laser. Soliton pairs and triplets of different types are found in parameter domain where single soliton mode-locking is unstable. Similar scenario is found for soliton pair and triplet: an additional weak soliton supplements already existing solitons, then with increasing gain the solitons start oscillating and at higher gain the soliton complex becomes stable. Spectral profiles of the solitons dynamically change taking either parabolic-like or Pi-like or M-like shape similar to those described in the literature for individual dissipative soliton at different pump level.

Properties and stability limits of an optimized mode-locked Yb-doped femtosecond fiber laser.

Stable mode-locked operation in a simple normal-cavity-dispersion laser oscillator that consists of only Yb-doped fiber and saturable absorber is studied. The Yb-doped fiber design parameters: group velocity dispersion (GVD), nonlinearity coefficient, bandwidth (Yb-BW), length and gain are considered to be the controlling parameters of the laser cavity. The pulse characteristics such as the temporal width, spectrum and pulse energy as a function of these elements are reported here. A pulse spectrum transition from M-like to Pi-like and then to parabolic-like shape is observed with different values of the controlling parameters which are similar to that has been observed before in a solid-state laser. The stability limits in the domain of the Yb-BW and length are studied. The stability dependence on GVD, nonlinearity coefficient and gain of the Yb-doped fiber are elucidated. Moreover, the pulse instability dyn ics beyond stability limits are found to be similar to that reported before for similariton.

Pulse compression and shaping of broadband optical parametric amplifier laser source.

We report pulse compression and shaping of a 100 Hz broadband optical parametric amplifier (OPA) laser source generated by self-phase modulation in a hollow-core fiber. The amplitude and phase of the broadband OPA laser pulses are controlled using an acousto optic programmable dispersive filter (AOPDF). Using the AOPDF, we demonstrate compression, characterization, and amplitude/phase control of 1300 nm 20 fs laser pulses with energies up to 10 microJ. This novel source is suitable for seeding successive OPA amplification stages and for time-resolved spectroscopy.

Third-order dispersion impact on mode-locking regimes of Yb-doped fiber laser with photonic bandgap fiber for dispersion compensation.

Novel features in stretched-pulse and similariton mode-locked regimes of Yb-doped fiber laser with photonic bandgap fiber used for dispersion compensation are found by means of numerical simulations. We show that the mode-locked pulse may become shorter with increasing third-order dispersion. Analytical estimations explain observed behavior through resonant interaction of the main pulse with dispersive waves involving both resonant sidebands and zero-group-velocity dispersion waves. Switching between the stretched-pulse and the similariton regimes is also studied.

Similariton pulse instability in mode-locked Yb-doped fiber laser in the vicinity of zero cavity dispersion.

Stability of the similariton mode-locked regime in Yb-doped fiber laser in the vicinity of zero cavity dispersion is studied by means of numerical simulations. It is shown that similariton pulses which initially arise from laser noise collapse into a continuous wave state. The mode-locked pulses are found to be stable after a certain cavity dispersion threshold is exceeded. From analysis of the instability development, we conclude that instability has parametric nature. We compare our results with stability analysis based on the Ginzburg-Landau approach. Analogies with instabilities found in the long-haul fiber communication systems are also discussed.

Coherent anti-Stokes Raman scattering microscopy using photonic crystal fiber with two closely lying zero dispersion wavelengths.

We demonstrate coherent anti-Stokes Raman scattering (CARS) microscopy of lipid-rich structures using a single unamplified femtosecond Ti:sapphire laser and a photonic crystal fiber (PCF) with two closely lying zero dispersion wavelengths (ZDW) for the Stokes source. The primary enabling factor for the fast data acquisition (84 micros per pixel) in the proof-of-principle CARS images, is the low noise supercontinuum (SC) generated in this type of PCF, in contrast to SC generated in a PCF with one ZDW. The dependence of the Stokes pulse on average input power, pump wavelength, pulse duration and polarization is experimentally characterized. We show that it is possible to control the spectral shape of the SC by tuning the pump wavelength of the input pulse and the consequence for CARS microscopy is discussed.