Severe photobleaching in two-photon excitation fluorescence microscopy (TPM) has limited its potential for long-term and quantitative imaging of live cells and tissues. One solution is to excite fluorophores with a high-repetition rate which reduces the peak intensity of irradiance. However, there is a lack of knowledge about the general utility of this strategy for fluorescent proteins. Here, using simple photobleaching assay, we studied the photobleaching of EGFP and tdTomato at 80 MHz and 640 MHz repetition rates. Our analysis shows that the impact of high repetition rate excitation on reducing photobleaching in TPM is highly dependent on the excitation wavelength and fluorophores. Our results are useful for selecting optimal parameters to minimize photobleaching and photodamage in TPM.

Point Spread Function (PSF) engineering is an effective method to increase the sensitivity of single-molecule fluorescence images to specific parameters. Classical phase mask optimization approaches have enabled the creation of new PSFs that can achieve, for example, localization precision of a few nanometers axially over a capture range of several microns with bright emitters. However, for complex high-dimensional optimization problems, classical approaches are difficult to implement and can be very time-consuming for computation. The advent of deep learning methods and their application to single-molecule imaging has provided a way to solve these problems. Here, we propose to combine PSF engineering and deep learning approaches to obtain both an optimized phase mask and a neural network structure to obtain the 3D position and 3D orientation of fixed fluorescent molecules. Our approach allows us to obtain an axial localization precision around 30 nanometers, as well as an orientation precision around 5 degrees for orientations and positions over a one micron depth range for a signal-to-noise ratio consistent with what is typical in single-molecule cellular imaging experiments.

Self-mixed optical feedback interferometry based laser sensors show promising results in the measurement of the vibration frequency. To date several measurement methods have been developed to extract the vibration information from the self-mixed (SM) signal; however, the complexity and accuracy of the methods still need improvement. The presented work tries to fulfill the gap by realizing a novel method using maximal overlap discrete wavelet transformation (MODWT) and multi-resolution analysis (MRA). The proposed method can reconstruct the micro-harmonic (< ) vibration up to 1 kHz even under weak feedback conditions. The mean squared error and the maximum relative error of the proposed method for this range remained below 1.89 & 8.79%, respectively. Although, above 1 kHz, the proposed method turns out to be futile to reconstruct the vibration signal but still capable to measure vibration frequency up to 10 kHz with an accuracy of 0.0001. The method also found suitable to measure non-sinusoidal vibration frequency with reasonable accuracy even for the moderate feedback conditions. The authors envision that the proposed method will provide a compact, non-contact, and low-cost alternative for the vibration frequency measurement hence useful in early fault detection schemes and lung abnormality diagnosis.

We demonstrate single-shot non-diffracting light-sheet generation by controlling the spatial coherence of light. A one-dimensional coherent beam, created by either increasing the spatial coherence of an LED or decreasing the spatial coherence of a laser, makes it unnecessary to scan non-diffracting beams to generate light-sheets. We theoretically and experimentally demonstrate the equivalence between our method and a scanned light-sheet, and investigate the characteristics of the light-sheet in detail. Our method is easily implementable and universally applicable for high-resolution multicolor light-sheet fluorescence imaging.

Tailoring electromagnetic waves has a wide range of applications, such as optical trapping, focus engineering, imaging, laser cutting, and optical communication. To do so, the spatial distribution of at least one of the four degrees of freedom of electromagnetic waves, amplitude, phase, polarization ratio, and retardance, must be modified. Arbitrary vectorial optical fields (VOF) can be engineered by spatially modulating all four degrees of freedom simultaneously. However, existing dynamic vectorial optical field generators (VOF-Gens) require intensive alignment and many optical elements in order to achieve high efficiency. Here, we design a dynamic VOF-Gen that can generate arbitrary VOFs using only five optical elements. Experimentally, we demonstrated an efficiency of 72%, the highest ever demonstrated.

We present a dual wavelength digital holographic technique for three-dimensional microscopic imaging of layered structures, where layers are separated from one another by the axial distances exceeding the wavelength of imaging light. Our methodology not only provides the three-dimensional structure of each layer, but also allows the height differentiation of distinct layers. We have also implemented a technique suppressing low intensity signal when no reliable phase information can be extracted, based on the quality of the interference fringe pattern. We utilize a dual wavelength setup, where the combination of two overlapping interferometers enables simultaneous acquisition of two phase profiles. We demonstrate that this imaging modality is particularly well-suited for imaging of multilayered electrode structures embedded in glass.

Patients with visual field loss have difficulty in mobility due to collision with pedestrians/obstacles from the blind side. In order to retrieve the lost visual field, prisms which deflect the field from the blind to the seeing side, have been widely used. However, the deflection power of current clinical Fresnel prisms is limited to ~30° and only provides a 5° eye scanning range to the blind side. This is not sufficient to avoid collision and results in increasing demands for a device with a higher power. In this paper, we propose a novel design and optimization of a higher power prism-like device (cascaded structure of mirror pairs filled with high refractive index) and verify enhanced expansion of up to 45° in optical ray tracing and photorealistic simulations.

We report the patterned synthesis of ZnO nanorod arrays of diameters between 50 nm and 130 nm and various spacings. This was achieved by patterning hole arrays in a polymethyl methacrylate layer with electron beam lithography, followed by chemical synthesis of ZnO nanorods in the patterned holes using the hydrothermal method. The fabrication of ZnO nanorod waveguide arrays is also demonstrated by embedding the nanorods in a silver film using the electroplating process. Optical transmission measurement through the nanorod waveguide arrays is performed and strong resonant transmission of visible light is observed. We have found the resonance shifts to a longer wavelength with increasing nanorod diameter. Furthermore, the resonance wavelength is independent of the nanowaveguide array period, indicating the observed resonant transmission is the effect of a single ZnO nanorod waveguide. These nanorod waveguides may be used in single-molecule imaging and sensing as a result of the nanoscopic profile of the light transmitted through the nanorods and the controlled locations of these nanoscale light sources.

We develop a point-spread function (PSF) engineering approach to imaging the spatial and spectral information of molecular emissions using a spatial light modulator (SLM). We show that a dispersive grating pattern imposed upon the emission reveals spectral information. We also propose a deconvolution model that allows the decoupling of the spectral and 3D spatial information in engineered PSFs. The work is readily applicable to single-molecule measurements and fluorescent microscopy.

Like all methods of super-resolution microscopy, stimulated emission depletion (STED) microscopy can suffer from the effects of aberrations. The most important aspect of a STED microscope is that the depletion focus maintains a minimum, ideally zero, intensity point that is surrounded by a region of higher intensity. It follows that aberrations that cause a non-zero value of this minimum intensity are the most detrimental, as they inhibit fluorescence emission even at the centre of the depletion focus. We present analysis that elucidates the nature of these effects in terms of the different polarisation components at the focus for two-dimensional and three-dimensional STED resolution enhancement. It is found that only certain low-order aberration modes can affect the minimum intensity at the Gaussian focus. This has important consequences for the design of adaptive optics aberration correction systems.

Lower coherence length and higher intensity are two indispensable requirements on the light source for high resolution and large penetration depth OCT imaging. While tremendous interest is being paid on engineering various laser sources to enlarge their bandwidth and hence lowering the coherence length, here we demonstrate another approach by employing strong temporal dispersion onto the existing laser source. Cholesteric liquid crystal (CLC) cells with suitable dispersive slope at the edge of 1-D organic photonic band gap have been designed to provide maximum reduction in coherence volume while maintaining the intensity higher than 50%. As an example, the coherence length of a multimode He-Ne laser is reduced by more than 730 times.

Based on an array detector, a new heterodyne detection system, which can correct the mismatches of amplitude and phase between signal and local oscillation (LO) beams, is presented in this paper. In the light of the fact that, for a heterodyne signal, there is a certain phase difference between the adjacent two samples of analog-to-digital converter (ADC), we propose to correct the spatial phase mismatch by use of the time-domain phase difference. The corrections can be realized by shifting the output sequences acquired from the detector elements in the array, and the steps of the shifting depend on the quantity of spatial phase mismatch. Numerical calculations of heterodyne efficiency are conducted to confirm the excellent performance of our system. Being different from previous works, our system needs not extra optical devices, so it provides probably an effective means to ease the problem resulted from the mismatches.

Conventional radiographic techniques depend on attenuation, which provides low contrast between soft tissues. However, X rays can accumulate large differential phase delays even in weakly absorbing materials. This can produce significantly higher contrast. One technique for taking advantage of phase effects, propagation-based phase imaging, can yield marked edge enhancement but requires spatially coherent intense sources. Microfocus sources have sizes on the order of tens of microns but necessarily are low power and hence require long exposures. In this project, X-ray optical and computational techniques were explored to develop both edge-enhancement and phase imaging using a large spot conventional source. A polycapillary optic was employed to create a small secondary source from a large spot rotating anode X-ray generator. The secondary spot created by the focusing polycapillary optic was 114 µm ± 50 µm. Images of a 1.6 mm polyethylene rod were taken at varying distances from the optic. Edge enhancement was observed with a maximum edge-enhancement-to-noise ratio of 6.5. Insect images were also acquired and analyzed. Phase reconstructions were computed using two different approaches, weak attenuation and phase attenuation duality. Pure phase images were successfully reconstructed from the phase contrast images by employing the weak attenuation model.

The generalised phase contrast (GPC) method provides versatile and efficient light shaping for a range of applications. We have implemented a generalised phase contrast system that used two passes on a single spatial light modulator (SLM). Both the pupil phase distribution and the phase contrast filter were generated by the SLM. This provided extra flexibility and control over the parameters of the system including the phase step magnitude, shape, radius and position of the filter. A feedback method for the on-line optimisation of these properties was also developed. Using feedback from images of the generated light field, it was possible to dynamically adjust the phase filter parameters to provide optimum contrast.

We present a digital holography microscopy technique based on parallel-quadrature phase-shifting method. Two π/2 phase-shifted holograms are recorded simultaneously using polarization phase-shifting principle, slightly off-axis recording geometry, and two identical CCD sensors. The parallel phase-shifting is realized by combining circularly polarized object beam with a 45° degree polarized reference beam through a polarizing beam splitter. DC term is eliminated by subtracting the two holograms from each other and the object information is reconstructed after selecting the frequency spectrum of the real image. Both amplitude and phase object reconstruction results are presented. Simultaneous recording eliminates phase errors caused by mechanical vibrations and air turbulences. The slightly off-axis recording geometry with phase-shifting allows a much larger dimension of the spatial filter for reconstruction of the object information. This leads to better reconstruction capability than traditional off-axis holography.

Based on a non-spherical model of particle scattering, we investigate the capabilities and limitations of a T-matrix based inverse algorithm to morphologically characterize cells in concentrated suspensions. Here the cells are modeled as randomly orientated spheroidal particles with homogenous dielectric properties and suspended in turbid media. The inverse algorithm retrieves the geometrical parameters and the concentration of cells simultaneously by inverting the reduced scattering coefficient spectra obtained from multispectral diffuse optical tomography (MS-DOT). Both round and spheroidal cells are tested and the role of multiple and higher order scattering of particles on the performance of the algorithm is evaluated using different concentrations of cells.

We present a robust encryption method for the encoding of 2D/3D objects using digital holography and virtual optics. Using our recently developed dual-plane in-line digital holography technique, two in-line digital holograms are recorded at two different planes and are encrypted using two different double random phase encryption configurations, independently. The process of using two mutually exclusive encryption channels makes the system more robust against attacks since both the channels should be decrypted accurately in order to get a recognizable reconstruction. Results show that the reconstructed object is unrecognizable even when the portion of the correct phase keys used during decryption is close to 75%. The system is verified against blind decryptions by evaluating the SNR and MSE. Validation of the proposed method and sensitivities of the associated parameters are quantitatively analyzed and illustrated.

Spectral domain phase microscopy (SDPM) is an extension of spectral domain optical coherence tomography (SDOCT) that exploits the extraordinary phase stability of spectrometer-based systems with common-path geometry to resolve sub-wavelength displacements within a sample volume. This technique has been implemented for high resolution axial displacement and velocity measurements in biological samples, but since axial displacement information is acquired serially along the lateral dimension, it has been unable to measure fast temporal dynamics in extended samples. Depth-Encoded SDPM (DESDPM) uses multiple sample arms with unevenly spaced common path reference reflectors to multiplex independent SDPM signals from separate lateral positions on a sample simultaneously using a single interferometer, thereby reducing the time required to detect unique optical events to the integration period of the detector. Here, we introduce DESDPM and demonstrate the ability to acquire useful phase data concurrently at two laterally separated locations in a phantom sample as well as a biological preparation of spontaneously beating chick cardiomyocytes. DESDPM may be a useful tool for imaging fast cellular phenomena such as nervous conduction velocity or contractile motion.

The use of scattered light images is shown to be an attractive method for the characterization of optofluidic waveguides. The method is shown to be capable of measuring waveguide propagation losses and transmissions between solid and liquid-core structures. Measurement uncertainties are considered and characterized and were typically less than 15%.

We demonstrate the first application of linear spectrogram methods based on electro-optic phase modulation to characterize optical arbitrary waveforms generated under spectral line-by-line control. This approach offers both superior sensitivity and self-referencing capability for retrieval of periodic high repetition rate optical arbitrary waveforms.

We explore the use of a switchable single-photon detector (SPD) array scheme to reduce the effect of a detector's deadtime for a multi-bit/photon quantum link. The case of data encoding using M possible orbital-angular-momentum (OAM) states is specifically studied in this paper. Our method uses SPDs with a controllable × optical switch and we use a Monte Carlo-based method to simulate the quantum detection process. The simulation results show that with the use of the switchable SPD array, the detection system can allow a higher incident photon rate than what might otherwise be limited by detectors' deadtime. For the case of = 4, = 20, a 50-ns deadtime for the individual SPDs, an average photon number per pulse of 0.1, and under the limit that at most 10 % of the photon-containing pulses are missed, the switchable SPD array will allow an incident photon rate of 2250 million counts/s (Mcts/s). This is 25 times the 90 Mcts/s incident photon rate that a non-switchable, 4-SPD array will allow. The increase in incident photon rate is more than the 5 times increase, which is the simple increase in the number of SPDs and the number of OAM encoding states (e.g., / = 20/4).