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Chondrocyte viability is a crucial factor in evaluating cartilage health. Most cell viability assays rely on dyes and are not applicable for in vivo or longitudinal studies. We previously demonstrated that two-photon excited autofluorescence and second harmonic generation microscopy provided high-resolution images of cells and collagen structure; those images allowed us to distinguish live from dead chondrocytes by visual assessment or by the normalized autofluorescence ratio. However, both methods require human involvement and have low throughputs. Methods for automated cell-based image processing can improve throughput. Conventional image processing algorithms do not perform well on autofluorescence images acquired by nonlinear microscopes due to low image contrast. In this study, we compared conventional, machine learning, and deep learning methods in chondrocyte segmentation and classification. We demonstrated that deep learning significantly improved the outcome of the chondrocyte segmentation and classification. With appropriate training, the deep learning method can achieve 90% accuracy in chondrocyte viability measurement. The significance of this work is that automated imaging analysis is possible and should not become a major hurdle for the use of nonlinear optical imaging methods in biological or clinical studies.

Laser cell patterning is a distinctly effective technique for creating cell arrangements in a culture that replicates in vivo tissue structure for studying contact-mediated cell-cell interactions. Conventional laser-based single-cell-manipulation techniques are limited by their inability to pattern irregularly shaped cells, such as rod-shaped cardiomyocytes. We report the use of a spatial light modulator loaded with a computer-generated phase map to shape a single laser source into multiple laser-guidance beams distributed around the outer contour of an irregularly shaped cell to achieve accurate cell patterning. In addition to describing the principle and practice of the system design, we present what is to our knowledge the first achievement of patterning large, irregularly (rod) shaped adult rat cardiomyocytes in an end-to-end connected alignment to replicate the in vivo heart muscle cell connection without the use of substrate surface modifications, which can interfere with in vivo-like cell-cell and cell-extracellular matrix interactions. Our research demonstrates that two-stage multiple-beam laser guidance is effective: 1) A dual-beam configuration horizontally translates a cell from the outlet of the microfluidic cell delivery channel to a position above the cell deposition site and 2) A quad-beam configuration rapidly propels the cell axially through the suspension medium to the culture substrate with the vertical movement of the cell patterning chamber. Our study reveals that 90% of the patterned cells maintained end-to-end connection 30 minutes after patterning, and mechanical junctions could be reinstalled between laser connected cells after overnight incubation. This demonstrates that multiple-beam laser patterning is an outstanding tool for in vitro studying contact-mediated cell-cell interactions among irregularly shaped cells.

Along with the recent surge in scale expansion of data centers, the interconnection scheme is facing a grave challenge. A huge amount of cables between the switches make the system maintenance and heat dissipation extremely difficult. A promising solution to this problem is using the arrayed waveguide grating (AWG), which can provide a set of wavelength links between its inputs and outputs. However, the scalability of the AWG-based interconnection scheme is restricted by the coherent crosstalk and the wavelength granularity of AWGs. In this paper, we propose a generic modular AWG-based interconnection scheme with scalable wavelength granularity for mega data centers. We first devise a matrix-based method to decompose the AWG into a three-stage network of smaller AWGs, while preserving the nonblocking wavelength routing property of the AWGs. We then introduce the concept of wavelength independency based on the partitioning of the optical connections, such that modular AWGs in the network can reuse the same wavelength set with smaller granularity. We show that the proposed modular AWG-based interconnection network can simplify the cabling complexity of data center networks, while preserving the same function and bandwidth as the original data center network.

Super-resolution microscopy (SRM) has had a substantial impact on the biological sciences due to its ability to observe tiny objects less than 200 nm in size. Stimulated emission depletion (STED) microscopy represents a major category of these SRM techniques that can achieve diffraction-unlimited resolution based on a purely optical modulation of fluorescence behaviors. Here, we investigated how the laser beams affect fluorescence lifetime in both confocal and STED imaging modes. The results showed that with increasing illumination time, the fluorescence lifetime in two kinds of fluorescent microspheres had an obvious change in STED imaging mode, compared with that in confocal imaging mode. As a result, the reduction of saturation intensity induced by the increase of fluorescence lifetime can improve the STED imaging resolution at the same depletion power. The phenomenon was also observed in Star635P-labeled human Nup153 in fixed HeLa cells, which can be treated as a reference for the synthesis of fluorescent labels with the sensitivity to the surrounding environment for resolution improvement in STED nanoscopy.

We propose an Edge-buffered Contention- resolution Hybrid Optical/Electrical Switch (ECHOES) for Data Center Networks. Simulations indicate our scheme can support a workload up to 0.93 while still maintain near 100% throughput and stable delay at microsecond-level.

With the rapid development of life sciences, there is an increasing demand for intravital fluorescence imaging of small animals. However, large dimensions and limited working distances of objective lenses in traditional fluorescence microscopes have limited their imaging applications mostly to superficial tissues. To overcome these disadvantages, researchers have developed the graded-index (GRIN) probes with small diameters for imaging internal organs of small animals in a minimally invasive fashion. However, dynamic imaging based on GRIN lens has not been studied extensively. Here, this paper presented a fluorescence endoscopic imaging system based on GRIN lenses using one-photon and two-photon excitation. GRIN lenses with 1.15 mm diameter and 7.65 mm length were used in the system. The images were acquired by a compact laser scanning imaging system with a resonant galvo-mirror system to scan the laser beam and a photomultiplier tube (PMT) to detect fluorescence signals. Experimental results showed that this system using two-photon excitation could implement dynamic fluorescence microendoscopic imaging and monitor the movement of blood flow beneath the skin in anesthetized mice while producing images with higher contrast and signal to noise ratio (SNR) than those using one photon excitation. It would be a useful tool for studying biological processes of small animals or plants in vivo.

Stimulated emission depletion microscopy (STED) is one of far-field optical microscopy techniques that can provide sub-diffraction spatial resolution. The spatial resolution of the STED microscopy is determined by the specially engineered beam profile of the depletion beam and its power. However, the beam profile of the depletion beam may be distorted due to aberrations of optical systems and inhomogeneity of specimens’ optical properties, resulting in a compromised spatial resolution. The situation gets deteriorated when thick samples are imaged. In the worst case, the sever distortion of the depletion beam profile may cause complete loss of the super resolution effect no matter how much depletion power is applied to specimens. Previously several adaptive optics approaches have been explored to compensate aberrations of systems and specimens. However, it is hard to correct the complicated high-order optical aberrations of specimens. In this report, we demonstrate that the complicated distorted wavefront from a thick phantom sample can be measured by using the coherent optical adaptive technique (COAT). The full correction can effectively maintain and improve the spatial resolution in imaging thick samples.

Objective To systematically evaluate the efficacy and safety of Modified Tongxie Yaofang (M-TXYF) for the treatment of diarrhea-predominant irritable bowel syndrome (IBS-D). Method Electronic databases including PubMed, Springer Link, EMBASE, China National Knowledge Infrastructure (CNKI), Chinese Biomedical Literature (CBM), Wanfang, and Chinese Scientific Journals Database (VIP) were conducted from their inception through May 11, 2017 without language restrictions. Primary and secondary outcomes were estimated by 95% confidence intervals (CI). RevMan 5.3 and the Cochrane Collaboration’s risk of bias tool were analyzed for this meta-analysis. Results Twenty-three literatures with a total of 1972 patients were included for the meta-analysis. The overall risk of bias evaluation was low. The pooled odds ratio showed that M-TXYF was significantly superior to routine pharmacotherapies (RP) in clinical therapeutic efficacy (OR 4.04, 95% CI 3.09, 5.27, P < 0.00001, therapeutic gain = 17.6%, number needed to treat (NNT) = 5.7). Moreover, compared with RP, M-TXYF showed that it can significantly reduce the scores of abdominal pain (standardized mean difference (SMD) -1.27; 95% CI -1.99, -0.56; P = 0.0005), abdominal distention (SMD -0.37; 95% CI -0.73, -0.01; P = 0.09), diarrhea (SMD -1.10; 95% CI -1.95, -0.25; P = 0.01), and frequency of defecation (SMD -1.42; 95% CI -2.19, -0.65; P = 0.0003). The differences of the adverse events between experiment and control groups had no statistical significance. Conclusion This meta-analysis indicated that M-TXYF could be a promising Chinese herbal formula in treating IBS-D. However, considering the lack of higher quality of randomized controlled trials (RCTs), highly believable evidences should be required.

We describe the design and implementation of two-photon lasers scanning stereomicroscopy, which allows viewing dynamic processes in three-dimensional (3D) space stereoscopically in real-time at the speed of 1.4 volumes per second.

This paper proposes an arrayed-waveguide grating (AWG) based wavelength-division-multiplexing (WDM) shuffle network. Compared with previous optical shuffle networks, our proposal is compact, easy to implement, highly scalable, and cost effective.

Multi-photon product (F540-state) of bacteriorhodopsin (BR) polymeric films excited by femtosecond laser shows permanently photoinduced birefringence and dichroism. Based on this property, polarization-sensitive diffractive optical elements (PDOEs), e.g., gratings, two-dimensional optical lattice, Bessel zone plate, were firstly fabricated and investigated in BR films by femtosecond laser multi-beam interference with rotationally symmetric pyramid lenses. These fabricated PDOEs are completely stable at room temperature. The dependence of diffraction efficiency on the incident polarization state was measured. The micro-structures of PDOEs were observed under polarization microscope, and the selective readout diffraction patterns from the polarization-multiplexed PDOEs were demonstrated.

Plasma enhanced ignition is an important way in the field of ignition auxiliary. It has got wide attention throughout the researchers both at home and abroad. Emission spectrum analysis is an important non-contact measurement method and could be used to analyze the parameters of the plasma. The coaxial corona discharge device was built and the emission spectrum was measured and analyzed. The physical model was constructed to calculate the vibrational temperature of nitrogen molecular. The results of the experiment and calculation results show that the emission spectrum of corona discharge is mainly second positive band system of nitrogen molecule. It is produced by the transition of the N2(C3Πu) and N2(B3Πu) which is the strongest and the most stable N2 emission spectrum. Ignore the emission spectrum of single line with no line width. The emission spectrum is mainly the N2(C-B,0-0), N2(C-B,1-3), N2(C-B,0-2), N2(C-B,1-4), N2(C-B,0-3) transition spectrum. The emission spectrum line of O2 is not obvious in the air corona discharge emission spectrum. The vibration temperature of nitrogen molecules was calculated according to the second positive band. The vibrational temperature of nitrogen molecules is 6193.21K, 3076.46K, 2788.61K at the discharge power of 1.325W, 4.272W, 8.4W. It laid a foundation of plasma enhance ignition for the further research.

Stimulated emission depletion microscopy (STED) has become one of the powerful research tools in the field of superresolution microscopy. As its spatial resolution is gained by phase modulation of the light field, the aberrations produced by optical systems and specimens may have negative impact on the focusing properties of two beams, especially the STED beam, resulting in reduced spatial resolution. In thick samples, the aberration effect may play an even more critical role in affecting spatial resolution. Here, we report our recent effort in correcting the aberration in STED microscopy by using coherent optical adaptive technique (COAT) so that the resolution can be improved.

An on-axis polarization phase-shifting digital holographic microscope, based on a normal upright optical microscope, is constructed to quantitatively measure both the amplitude and phase distributions of specimen. The condenser lens and microscope objective employed in the object beam path enhance the illumination and magnification of the image, however, they induce additional phase aberration of the object wave. The physical formation of the phase aberration is theoretically analyzed, and a formula for the object wave front involving the phase aberration in the CCD plane is derived. The phase aberration can be eliminated in the reconstruction procedure by measuring a specimen-free hologram and then fitting the aberration phase with a least square ellipsoidal model to determine the parameters of the system. This phase aberration compensation procedure also reduces some of noises in the reconstructed phase of the specimen. The practicability of this method is demonstrated by a test experiment on microlenses.

Dual-channel phase-shifting interferometry for simultaneous phase microscopy is presented. Red and blue light beams are used for microscope illumination. A 45° tilted beamsplitter replicates the object and reference waves in red light together with the object wave in blue light into two parallel beams. The two resulting quadrature phase-shifting interferograms in red light and the object waves in blue light are generated in the two channels. The two interferograms are recorded simultaneously by a color charge-coupled device (CCD) camera, and can be separated via RGB components of the recorded color patterns without crosstalk. As a result, the phase of tested specimen can be retrieved. The feasibility of the proposed method is demonstrated by test performed on a microscopic specimen.

Parallel two-step phase-shifting interferometry for microscopy is presented, and the recording condition for generalized two-step phase-shifting interferometry is discussed. A 45° tilted cube beamsplitter enables to replicate the orthogonally linear polarized object and reference waves into two parallel beams, respectively. As a consequence, two interferograms with quadrature phase shift are obtained along the two beams, and phase reconstructed with an improved algorithm. To reconstruct the phase distribution from the two-step phase-shifting interferograms, a certain recording condition should be satisfied. However, the recording condition has not ever been discussed before. In this paper, the recording condition for the two-step phase-shifting interferometry is derived and that is: the intensity of reference wave should be no less than two times object wave intensity.

A novel technique, based on electron-beam lithography (EBL) and anisotropic reactive ion etch (RIE), was developed in this work for the fabrications of single electron transistors with ultra small junctions in silicon on insulator (SOI). Junction size as small as 10nm has been achieved. Initial characterisation at low temperature has demonstrated clear Coulomb blockade gap in the measured I-V curves, indicating that such narrow junctions in silicon finally formed by RIE are still functional. This process eases the difficulty in EBL for 10nm feature size by the controllable lateral dry-etch on the sidewall of the silicon thin layer, opening up a prospect for routine manufacture of single electronic devices and circuits on SOI, which possesses broad applications in meteorological measurement and IT technology.

Semi-metals might offer a stronger interaction and a better confinement for terahertz wave than semiconductors, while preserve tunability. Particularly, graphene-based materials are envisioned as terahertz modulators, filters and ultra-broadband sources. However, the understanding of terahertz generation from those materials is still not clear, thus limits us recognizing the potential and improving device performances. Graphite, the mother material of graphene and a typical bulk semi-metal, is a good system to study semi-metals and graphene-based materials. Here we experimentally modulate and maximize the terahertz signal from graphite surface, thus reveal the mechanism - surface field driving photon induced carriers into transient current to radiate terahertz wave. We also discuss the differences between graphite and semiconductors; particularly graphite shows very weak temperature dependency from room temperature to 80 °C. Above knowledge will help us understand terahertz generations, achieve maximum output and electric modulation, in semi-metal or graphene based devices.

Bessel beams have been used in many applications due to their unique optical properties of maintaining their intensity profiles unchanged during propagation. In imaging applications, Bessel beams have been successfully used to provide extended focuses for volumetric imaging and uniformed illumination plane in light-sheet microscopy. Coupled with two-photon excitation, Bessel beams have been successfully used in realizing fluorescence projected volumetric imaging. We demonstrated previously a stereoscopic solution-two-photon fluorescence stereomicroscopy (TPFSM)-for recovering the depth information in volumetric imaging with Bessel beams. In TPFSM, tilted Bessel beams were used to generate stereoscopic images on a laser scanning two-photon fluorescence microscope; upon post image processing we could successfully provide 3D perception of acquired volume images by wearing anaglyph 3D glasses. However, tilted Bessel beams were generated by shifting either an axicon or an objective laterally; the slow imaging speed and severe aberrations made it hard to use in real-time volume imaging. In this article, we report recent improvements of TPFSM with newly designed scanner and imaging software, which allows 3D stereoscopic imaging without moving any of the optical components on the setup. This improvement has dramatically improved focusing qualities and imaging speed so that the TPFSM can be performed potentially in real-time to provide 3D visualization in scattering media without post image processing.

Bessel beams have been used in many applications due to their unique optical properties of maintaining their intensity profiles unchanged during propagation. In imaging applications, Bessel beams have been successfully used to provide extended focuses for volumetric imaging and uniformed illumination plane in light-sheet microscopy. Coupled with two-photon excitation, Bessel beams have been successfully used in realizing fluorescence projected volumetric imaging. We demonstrated previously a stereoscopic solution–two-photon fluorescence stereomicroscopy (TPFSM)–for recovering the depth information in volumetric imaging with Bessel beams. In TPFSM, tilted Bessel beams were used to generate stereoscopic images on a laser scanning two-photon fluorescence microscope; upon post image processing we could successfully provide 3D perception of acquired volume images by wearing anaglyph 3D glasses. However, tilted Bessel beams were generated by shifting either an axicon or an objective laterally; the slow imaging speed and severe aberrations made it hard to use in real-time volume imaging. In this article, we report recent improvements of TPFSM with newly designed scanner and imaging software, which allows 3D stereoscopic imaging without moving any of the optical components on the setup. This improvement has dramatically improved focusing qualities and imaging speed so that the TPFSM can be performed potentially in real-time to provide 3D visualization in scattering media without post image processing.

With the rapid development of life sciences, there is an increasing demand for intravital fluorescence imaging of small animals. However, large dimensions and limited working distances of objective lenses in traditional fluorescence microscopes have limited the imaging applications mostly to superficial tissues. To overcome this disadvantage, researchers have developed the graded-index (GRIN) probes with small diameters for imaging internal organs of small animals in a minimally invasive fashion. Here, we present the development of a fluorescence endoscopic imaging system based on a GRIN lens using two-photon excitation. Experimental results showed that this system could perform dynamic fluorescence microendoscopic imaging and monitor the blood flow in anesthetized living mice using two-photon excitation.

This paper studies the routing problems in Internet protocol/wavelength-division-multiplexing (IP/WDM) networks based on the overlay routing algorithm (ORA) and the integrated routing algorithm (IRA), respectively. Although IRA usually outperforms ORA in terms of blocking performance, IRA exhibits disadvantages in control information exchange, network privacy issue, and wavelength port efficiency. In this paper, a new mechanism called the short lightpath establishment approach (SLEA) is proposed for ORA in order to tackle the problems in IRA and achieve similar (or even better) network performance at the same time. The main idea of SLEA is to ensure that each new lightpath created by ORA is restricted by an optical hop constraint when a subwavelength-granularity connection is routed in the optical layer. It follows that SLEA essentially avoids per-connection-based greedy treatment and improves network wide resource utilization by eliminating inefficient long optical bypasses. To implement SLEA in ORA, the Dijsktra's algorithm has been modified based on an extended layered graph model. SLEA does not introduce any additional signaling and computational complexity. The analysis and simulation in this paper show that there exists an optimal optical hop constraint for each particular network configuration such that SLEA-based ORA (SLEA-ORA) can efficiently utilize the network resource of concern. As a result, with the optimal optical hop constraint, SLEA-ORA could outperform ORA and IRA in terms of the bandwidth-blocking ratio (BBR) and the average number of IP hops of label-switched paths (LSPs).

This paper focuses on dynamic integrated routing in multifiber Internet protocol/wavelength-division multiplexing (IP/WDM) networks, which can be implemented through either one-step routing (OSR) or two-step routing (TSR) approach. Based on an extended layered-graph, two resource assignment strategies, termed channel-level balance (CLB) and link-level balance (LLB), are proposed to balance the traffic in the network at different levels. To further improve the performance, a parameter K is introduced to make a dynamic tradeoff between the logical-layer links and the optical-layer links. Simulation studies are carried out for various topologies. The results show that LLB is better than CLB in most cases, and LLB combined with OSR has the optimal performance. Also, we find that the routing approach and the resource assignment strategy individually play different roles with different values of r/sub l/ that is introduced to indicate the resource richness of the network. As a multifiber network is functionally equivalent to a single-fiber network with limited wavelength conversion, we investigate the effects of wavelength conversion by studying the multifiber IP/WDM networks. The analysis shows that, when the granularity of each connection request is much smaller than the wavelength granularity, wavelength conversion may increase the request blocking probability in the network.

Jianpi Yiqi therapy (JYT) is a classical therapy in treating chronic atrophic gastritis (CAG), but the clinical effects of it are still contentious. The purpose of this article is to evaluate the efficacy and safety of JYT for CAG. Seven electronic databases including PubMed, Embase, Springer Link, CNKI (China National Knowledge Infrastructure), VIP (Chinese Scientific Journals Database), Wan-fang database, and CBM (Chinese Biomedicine Database) were searched from their inception to November 1, 2016. 13 randomized controlled trials (RCTs) with a total of 1119 participants were identified for analysis. Meta-analyses demonstrated that both JYT (RR 1.41; 95% CI 1.27, 1.57; P < 0.00001) and JYT + western medicine (RR 1.27; 95% CI 1.17, 1.38; P < 0.00001) were more efficacious than only western medicine. Furthermore, JYT had potential improvement on traditional Chinese medicine (TCM) symptoms scores such as stomachache, stomach distention, belching, fatigue, et al. In addition, no serious adverse events were reported in the selected trials. The Cochrane Collaboration's risk of bias tool was evaluated for the weaknesses of methodological quality, while the quality level of Grades of Recommendations Assessment Development and Evaluation (GRADE) evidence classification indicated "Very low". This meta-analysis indicates that JYT may have potential effects on the treatment of patients with CAG. However, due to limitations of methodological quality and small sample size of the included studies, further standardized research of rigorous design should be needed.

An on-axis phase-shifting reflective point-diffraction microscopic interferometer for quantitative phase microscopy based on Michelson architecture is proposed. A cube beamsplitter splits the object wave spectrum into two copies within two arms. Reference wave is rebuilt in one arm by low-pass filtering on the object wave frequency spectrum with a pinhole-mask mirror, and interferes with the object wave from the other arm. Polarization phase-shifting is performed and phase imaging on microscale specimens is implemented. The experimental results demonstrate that the proposed scheme has the advantage of long-term stability due to its quasi common-path geometry with full use of laser energy.

Remote-focusing microscopy has recently attracted a lot of interest due to its high-speed axial scanning capabilities. In this paper, we modeled remote-focusing microscopy, based on a pair of long working distance objective lenses. Three-dimensional intensity distributions of the point spread functions (PSFs) are calculated, and no significant spherical aberrations are introduced over a large volume of 100 μm×100 μm×150 μm. The validity of the scheme is verified by imaging biological samples and microelectronic chips at the imaging depth of 150 μm without introducing aberrations in the experiment.

We propose and demonstrate a tunable broadband photonic radio frequency (RF) phase shifter based on a silicon microring resonator. This scheme utilizes the thermal nonlinear effect of the silicon microring to change the electrical phase of the RF signal with a wide tuning range. A prototype of the phase shifter is experimentally demonstrated for a 40-GHz signal with a 0-4.6-rad tuning range.

We propose and experimentally demonstrate the generation of baseband ultra-wideband (UWB) monocycle and doublet pulses using one dual-parallel Mach-Zehnder modulator. We further present a proof-of-concept demonstration of a 24-GHz UWB over fiber system based on frequency up-conversion. The performance of the up-converted UWB pulses after fiber transmission is studied.

A reflection mode digital holographic microscope with light emitting diode (LED) illumination and off-axis interferometry is proposed. The setup is comprised of a Linnik interferometer and a grating-based 4f imaging unit. Both object and reference waves travel coaxially and are split into multiple diffraction orders in the Fourier plane by the grating. The zeroth and first orders are filtered by a polarizing array to select orthogonally polarized object waves and reference waves. Subsequently, the object and reference waves are combined again in the output plane of the 4f system, and then the hologram with uniform contrast over the entire field of view can be acquired with the aid of a polarizer. The one-shot nature in the off-axis configuration enables an interferometric recording time on a millisecond scale. The validity of the proposed setup is illustrated by imaging nanostructured substrates, and the experimental results demonstrate that the phase noise is reduced drastically by an order of 68% when compared to a He–Ne laser-based result.

Parallel on-axis two-step phase-shifting reflective point-diffraction interferometry for holographic phase microscopy based on Michelson architecture is proposed. A cube beamsplitter splits the object wave into two copies within the two arms. The reference wave is rebuilt by low-pass filtering with a pinhole-masked mirror. Both object and reference waves are split into two beams by a grating in a 4f imaging system; thus, two interferograms with quadrature phase-shift can be acquired simultaneously with the aid of polarization elements. The approach has the merit of nanometers-scale phase stability over hours due to its quasi-common-path geometry. It can make full use of camera spatial bandwidth while its temporal resolution is as fast as the camera frame rate. Phase imaging on microscale specimen is implemented, and the experimental results demonstrate that the proposed approach is suitable for investigating dynamic processes.

Three dimensional distributions of cells can be usually acquired by optical sectioning methods, such as multiphoton excitation and confocal fluorescence laser scanning microscopy. Though the lateral scan rates can reach up to several kHz, the relatively slow axial scan comprises the speed of real-time imaging of a volume. Here we propose a three dimensional imaging method that uses Bessel beams as excitation in multiphoton fluorescence microscopy. The extended focus of the Bessel beam allows recording a volume of cells without scanning the depth. The depth information can be retrieved by recording a pair of parallax views of the same volume. We have demonstrated the stereoscope capability on a homebuilt two-photon fluorescence microscope.

Super-resolution three-dimensional (3D) optical microscopy has incomparable advantages over other high-resolution microscopic technologies, such as electron microscopy and atomic force microscopy, in the study of biological molecules, pathways and events in live cells and tissues. We present a novel approach of structured illumination microscopy (SIM) by using a digital micromirror device (DMD) for fringe projection and a low-coherence LED light for illumination. The lateral resolution of 90 nm and the optical sectioning depth of 120 μm were achieved. The maximum acquisition speed for 3D imaging in the optical sectioning mode was 1.6×10(7) pixels/second, which was mainly limited by the sensitivity and speed of the CCD camera. In contrast to other SIM techniques, the DMD-based LED-illumination SIM is cost-effective, ease of multi-wavelength switchable and speckle-noise-free. The 2D super-resolution and 3D optical sectioning modalities can be easily switched and applied to either fluorescent or non-fluorescent specimens.

Super-resolution optical microscopy has some incomparable comprehensive advantages over various other microscopic technologies such as electron microscopy, atomic force microscopy and so on, especially in the study of biological molecules, pathways and events in live cells and tissues. In this report, we present a novel structured illumination microscopy (SIM) based on digital micromirror device (DMD) fringe projection with the low-coherence LED light as illumination source. A 90nm in-plane resolution was achieved with gold nano-particles and BPAE cells.

Bessel beams have been successfully used in two-photon laser scanning fluorescence microscopy to extend the depth of field (EDF), which makes it possible to observe fast events volumetrically. However, the depth information is lost due to integration of fluorescence signals along the propagation direction. We describe the design and implementation of two-photon lasers scanning stereomicroscopy, which allows viewing dynamic processes in three-dimensional (3D) space stereoscopically in real-time with shutter glasses at the speed of 1.4 volumes per second. The depth information can be appreciated by human visual system or be recovered with correspondence algorithms for some cases.

An autofocusing method for two-wavelength digital holographic microscopy (TWDHM) based on the wavelength dependence of the diffraction process is proposed. Red and green lights are employed for the illumination of the TWDHM, and the generated holograms are recorded simultaneously by a color CCD camera. Due to the wavelength dependency of the diffraction process, the farther the reconstruction plane is from the image plane, the larger the difference is between the red and green light distributions. Thus, the image plane can be determined by finding the minimum of the variation between the red and green lights on their amplitude distributions. The feasibility of the proposed method is demonstrated by simulation and experiment.

We present a stimulated emission depletion microscopy system that has been built around a single ultrafast laser system. Excitation beams are provided by a supercontinuum generation with the same laser.

We developed a hybrid two-photon excitation fluorescence-second harmonic generation (TPEF-SHG) imaging system with an on-stage incubator for long-term live-cell imaging. Using the imaging system, we observed the addition of new sarcomeres during myofibrillogenesis while a cardiomyocyte was spreading on the substrate. The results suggest that the TPEF-SHG imaging system with an on-stage incubator is an effective tool for investigation of dynamic myofibrillogenesis.

We developed a hybrid SHG-TPEF polarization imaging system that allowed the excitation beam from an fs Ti:Sappire laser being bi-directionally raster scanned across the focal plane using a pair of orthogonal galvanometers. To implement high-speed scanning, the turning regions of the triangular waves were smoothed by a custom-designed waveform. The SHG and TPEF signals from samples were recorded by two PMTs in the forward and backward direction. Using this imaging system, we obtained 3D images of the sarcomere structure via SHG and DiO-stained lipid membrane via TPEF in live cardiomyocytes isolated from neonatal and adult rats. The results demonstrated the potential applications of SHG and TPEF in the research of myofibrillogensis.

Continuously-tunable slow and fast light is experimentally demonstrated using silicon microring add-drop filter with mutual mode coupling. Pulse delay and advancement can be achieved by thermal tuning at the drop port of the resonator.

We propose a scheme to achieve regenerative format conversion from a distorted non-return-to-zero-on–off-keying (NRZ-OOK) signal to a return-to-zero-binary-phase-shift-keying (RZ-BPSK) signal of high quality in an integrated device. This proposal is achieved by employing an interferometer on a silicon chip with a coupled ring resonator optical waveguide embedded in one arm. In this scheme the amplified NRZ-OOK signal is injected into the converter together with a locally generated return-to-zero (RZ) pulse train at a different wavelength. Owing to the linear and nonlinear effects in the silicon waveguides, the RZ pulse train changes to a clean RZ-BPSK signal after it is cross modulated by the NRZ-OOK signal. To verify the feasibility of our scheme, we simulate the conversion at 10 Gbits/s.

Ultrafast laser pulse shaping makes it possible to observe nonlinear signatures, such as self phase modulation and sum frequency absorption, at safe laser powers for tissue. Neuronal activation and melanoma diagnosis are two early targets.

An optical transmitter for generating phase-shift-keying (PSK) radio-frequency (RF)-signal is experimentally demonstrated, where a high-frequency RF-carrier is obtained by self-heterodyning and the PSK-modulation is achieved by the amplitude-shift-keying-to-PSK conversion in an integrated dual-parallel Mach-Zehnder modulator.

Steady-state absorption and emission spectroscopies, oxygen activation and transient spectroscopy on a single sample of synthetic pheomelanin are compared. The absorption, emission and excitation spectra of pheomelanin depend on the molecular weight (MW) of the dissolved pigment constituents. While weakly-depending on MW, the maximum of the emission excitation spectrum is approximately 400 nm. The electron paramagnetic resonance oximetry measurements show a clear increase in oxygen uptake between 338 and 323 nm, and also reveal a local enhancement around approximately 370 nm. Pump-probe absorption spectroscopy reveals that photoexcitation of pheomelanin by UVA light generates a transient absorption peak in the visible and UV regions within the instrument response. The action spectrum for the formation of the 780 nm transient species peaks at approximately 360 nm. While both transient absorption bands show the same ultrafast decay component, the 780 nm peak completely vanishes on the 10s of picosecond time scale, but the UV band shows a kinetic evolution to a subsequent intermediate. While in a similar wavelength range, the maximum of the action spectrum derived from the transient data, the emission excitation spectrum and the action spectrum for photoconsumption all differ from one another, suggesting that the chromophore responsible for each is not that same. This raises concern about comparing the results from different photochemical methodologies for melanin, which is a specific case of comparing data on systems where molecular constituents are not well defined.

Even though multi-photon fluorescence microscopy offers higher resolution and better penetration depth than traditional fluorescence microscopy, its use is restricted to the detection of molecules that fluoresce. Two-photon absorption (TPA) imaging can provide contrast in non-fluorescent molecules while retaining the high resolution and sectioning capabilities of nonlinear imaging modalities. In the long-wavelength water window, tissue TPA is dominated by the endogenous molecules melanin and hemoglobin with an almost complete absence of endogenous two-photon fluorescence. A complementary nonlinear contrast mechanism is self-phase modulation (SPM), which can provide intrinsic signatures that can depend on local tissue anisotropy, chemical environment, or other structural properties. We have developed a spectral hole refilling measurement technique for TPA and SPM measurements using shaped ultrafast laser pulses. Here we report on a microscopy setup to simultaneously acquire 3D, high-resolution TPA and SPM images. We have acquired data in mounted B16 melanoma cells with very modest laser power levels. We will also discuss the possible application of this measurement technique to neuronal imaging. Since SPM is sensitive to material structure we can expect SPM properties of neurons to change during neuronal firing. Using our hole-refilling technique we have now demonstrated strong novel intrinsic nonlinear signatures of neuronal activation in a hippocampal brain slice. The observed changes in nonlinear signal upon collective activation were up to factors of two, unlike other intrinsic optical signal changes on the percent level. These results show that TPA and SPM imaging can provide important novel functional contrast in tissue using very modest power levels suitable for in vivo applications.

We developed two methods to access novel nonlinear functional contrast in tissue. Here we present three-dimensional, high-resolution data of two-photon absorption, sum-frequency absorption and self-phase modulation acquired in mounted melanoma cells. This fundamentally new contrast, combined with the technique’s high sensitivity, promises to significantly advance optical methods for tissue characterization and diagnosis.

Optical networks have been introduced to meet the bandwidth requirement of mega data centers (DC). Most existing approaches are neither scalable to face the massive growth of DCs, nor contention-free enough to provide full bisection bandwidth. To solve this problem, we propose two symmetric network structures: ring-MEMS-ring (RMR) network and MEMS-ring-MEMS (MRM) network based on classical Clos theory. New strategies are introduced to overcome the additional wavelength constraints that did not exist in the traditional Clos network. Two structures that followed the strategies can enable high scalability and nonblocking property simultaneously. The one-to-one correspondence of the RMR and MRM structures to a Clos is verified and the nonblocking conditions are given along with the routing algorithms. Compared to a typical folded-Clos network, both structures are more readily scalable to future mega data centers with 51200 racks while reducing number of long cables significantly. We show that the MRM network is more cost-effective than the RMR network, since the MRM network does not need tunable lasers to achieve nonblocking routing.