Rapid profiling of fish cell nitrogen metabolism with single-cell Raman spectroscopy: Unveiling enzyme's role in ammonia detoxification.

Ammonia is a prevalent aquatic pollutant that disrupts cellular functions and energy metabolism in fish, posing significant environmental and health threats. This research investigates the critical role of arginase 2 (ARG2) in mitigating ammonia toxicity in fish cells and its implications in adapting to nitrogen metabolism under high ammonia exposure. Through a CRISPR-Cas9 engineered ARG2 knockdown (KD) in the Epithelioma Papulosum Cyprini (EPC) cell line, we first investigated the biochemical responses of ARG2 KD and wild-type (WT) EPC cells to ammonia stress (NH4Cl treatment), showing diminished urea production and decreased cell viability in ARG2 KD cells. Subsequently, single-cell Raman spectroscopy analysis revealed that ARG2 KD cells exhibited profound metabolic shifts, including changes in protein, nucleic acids, lipid and sugar levels, showing the adjusting role of ARG2 in the balance of carbohydrate and nitrogen metabolism. Furthermore, the upregulated responses of various amino acids, such as glutamine, arginine, alanine, glutamic acid, glycine, histidine, phenylalanine and valine, in WT cells after NH4Cl treatment diminished in ARG2 KD cells except for the decrease in aspartic acid, indicating a switching effect of ARG2 in nitrogen metabolism under ammonia stress. This study highlights ARG2's essential role in ammonia detoxification and emphasizes ARG2's protective function and its importance in metabolism, shedding light on the adaptive mechanisms fish cells deploy against high ammonia environments. These insights contribute to deep understanding of aquatic organisms' molecular responses to environmental ammonia pollution, offering potential strategies for their protection.

Dispersive Fourier transform based dual-comb ranging.

Laser-based light detection and ranging (LIDAR) offers a powerful tool to real-timely map spatial information with exceptional accuracy and owns various applications ranging from industrial manufacturing, and remote sensing, to airborne and in-vehicle missions. Over the past two decades, the rapid advancements of optical frequency combs have ushered in a new era for LIDAR, promoting measurement precision to quantum noise limited level. For comb LIDAR systems, to further improve the comprehensive performances and reconcile inherent conflicts between speed, accuracy, and ambiguity range, innovative demodulation strategies become crucial. Here we report a dispersive Fourier transform (DFT) based LIDAR method utilizing phase-locked Vernier dual soliton laser combs. We demonstrate that after in-line pulse stretching, the delay of the flying pulses can be identified via the DFT-based spectral interferometry instead of temporal interferometry or pulse reconstruction. This enables absolute distance measurements with precision starting from 262 nm in single shot, to 2.8 nm after averaging 1.5 ms, in a non-ambiguity range over 1.7 km. Furthermore, our DFT-based LIDAR method distinctly demonstrates an ability to completely eliminate dead zones. Such an integration of frequency-resolved ultrafast analysis and dual-comb ranging technology may pave a way for the design of future LIDAR systems.

Beam-steering efficiency optimization method based on a rapid-search algorithm for liquid crystal optical phased array.

A rapid-search algorithm to improve the beam-steering efficiency for a liquid crystal optical phased array was proposed and experimentally demonstrated in this paper. This proposed algorithm, in which the value of steering efficiency is taken as the objective function and the controlling voltage codes are considered as the optimization variables, consisted of a detection stage and a construction stage. It optimized the steering efficiency in the detection stage and adjusted its search direction adaptively in the construction stage to avoid getting caught in a wrong search space. Simulations had been conducted to compare the proposed algorithm with the widely used pattern-search algorithm using criteria of convergence rate and optimized efficiency. Beam-steering optimization experiments had been performed to verify the validity of the proposed method.

Experimental generation of complex optical fields for diffraction limited optical focus with purely transverse spin angular momentum.

We demonstrate a method to generate complex optical fields at the pupil plane of a high numerical aperture (NA) objective lens for the creation of diffraction limited optical focus with purely transverse spin angular momentum. The complex optical fields are analytically deduced through reversing the radiated patterns from two electric dipoles, which are located at the focal point of the high NA lens and oscillate respectively in x- and z- directions with phase difference of π/2. The derived fields can be experimentally created with a vectorial optical field generator. Using the Richard-Wolf vectorial diffraction theory, the electric fields within the focal region are calculated to evaluate the intensities and polarization distributions of the tightly focused beams corresponding to both the theoretical and experimentally generated pupil fields and the results clearly demonstrate the validity of the proposed technique.

Precise transverse alignment of spatial light modulator sections for complex optical field generation.

Based on the properties of the dove prism and the Fourier optics approach, the coordinate relationships among four spatial light modulator (SLM) sections in a vectorial optical field generator are derived and experimentally verified. Taking the coordinate system of the first SLM section as a reference, the coordinate displacements between the first section and subsequent sections are determined via employing specially designed four-quadrant patterns, which enable the visualization of the degree of freedom controlled by each SLM section. A complex optical field could be accurately generated through combining the derived coordinate relationships and pre-compensation of the measured coordinate displacements. Several typical complex optical fields are experimentally generated to demonstrate the validity of the proposed transverse alignment method.

Demonstration of a vectorial optical field generator with adaptive close loop control.

We experimentally demonstrate a vectorial optical field generator (VOF-Gen) with an adaptive close loop control. The close loop control capability is illustrated with the calibration of polarization modulation of the system. To calibrate the polarization ratio modulation, we generate 45° linearly polarized beam and make it propagate through a linear analyzer whose transmission axis is orthogonal to the incident beam. For the retardation calibration, circularly polarized beam is employed and a circular polarization analyzer with the opposite chirality is placed in front of the CCD as the detector. In both cases, the close loop control automatically changes the value of the corresponding calibration parameters in the pre-set ranges to generate the phase patterns applied to the spatial light modulators and records the intensity distribution of the output beam by the CCD camera. The optimized calibration parameters are determined corresponding to the minimum total intensity in each case. Several typical kinds of vectorial optical beams are created with and without the obtained calibration parameters, and the full Stokes parameter measurements are carried out to quantitatively analyze the polarization distribution of the generated beams. The comparisons among these results clearly show that the obtained calibration parameters could remarkably improve the accuracy of the polarization modulation of the VOF-Gen, especially for generating elliptically polarized beam with large ellipticity, indicating the significance of the presented close loop in enhancing the performance of the VOF-Gen.