Spatiotemporally mapping temperature dynamics of lysosomes and mitochondria using cascade organelle-targeting upconversion nanoparticles.

The intracellular metabolism of organelles, like lysosomes and mitochondria, is highly coordinated spatiotemporally and functionally. The activities of lysosomal enzymes significantly rely on the cytoplasmic temperature, and heat is constantly released by mitochondria as the byproduct of adenosine triphosphate (ATP) generation during active metabolism. Here, we developed temperature-sensitive LysoDots and MitoDots to monitor the in situ thermal dynamics of lysosomes and mitochondria. The design is based on upconversion nanoparticles (UCNPs) with high-density surface modifications to achieve the exceptionally high sensitivity of 2.7% K-1 and low uncertainty of 0.8 K for nanothermometry to be used in living cells. We show the measurement is independent of the ion concentrations and pH values. With Ca2+ ion shock, the temperatures of both lysosomes and mitochondria increased by ∼2 to 4 °C. Intriguingly, with chloroquine (CQ) treatment, the lysosomal temperature was observed to decrease by up to ∼3 °C, while mitochondria remained relatively stable. Lastly, with oxidative phosphorylation inhibitor treatment, we observed an ∼3 to 7 °C temperature increase and a thermal transition from mitochondria to lysosomes. These observations indicate different metabolic pathways and thermal transitions between lysosomes and mitochondria inside HeLa cells. The nanothermometry probes provide a powerful tool for multimodality functional imaging of subcellular organelles and interactions with high spatial, temporal, and thermal dynamics resolutions.

Image Stitching Based on Color Difference and KAZE with a Fast Guided Filter.

Image stitching is of great importance for multiple fields such as moving object detection and tracking, ground reconnaissance and augmented reality. To ameliorate the stitching effect and alleviate the mismatch rate, an effective image stitching algorithm based on color difference and an improved KAZE with a fast guided filter is proposed. Firstly, the fast guided filter is introduced to reduce the mismatch rate before feature matching. Secondly, the KAZE algorithm based on improved random sample consensus is used for feature matching. Then, the color difference and brightness difference of the overlapping area are calculated to make an overall adjustment to the original images so as to improve the nonuniformity of the splicing result. Finally, the warped images with color difference compensation are fused to obtain the stitched image. The proposed method is evaluated by both visual effect mapping and quantitative values. In addition, the proposed algorithm is compared with other current popular stitching algorithms. The results show that the proposed algorithm is superior to other algorithms in terms of the quantity of feature point pairs, the matching accuracy, the root mean square error and the mean absolute error.

Modified Two-Point Correction Method for Wide-Spectrum LWIR Detection System.

Non-uniformity commonly exists in the infrared focal plane, which behaves as the fixed-pattern noise (FPN) and seriously affects the image quality of long-wave infrared (LWIR) detection systems. The two-point correction (TPC) method is commonly used to reduce image FPN in engineering. However, when a wide-spectrum LWIR detection system calibrated with a black body is used to detect weak and small targets in the sky, FPN still appears in the image, affecting its uniformity. The effects of atmospheric transmittance characteristics of long-range paths on the non-uniformity of wide-spectrum long-wave infrared systems have not been studied. This paper proposes a modified TPC model based on spectral subdivision that introduces atmospheric transmittance. Additionally, the effects of atmospheric transmittance characteristics on the long-wave infrared non-uniform correction coefficient are analyzed. The experimental results for a black body scene and sky scene using a weak and small target detection system with a long-wave Sofradir FPA demonstrate that the wide-spectrum LWIR detection system fully considers atmospheric transmittance when performing calibration based on the TPC method, which can reduce the non-uniformity of the image.

Single Small Extracellular Vesicle (sEV) Quantification by Upconversion Nanoparticles.

Cancer-derived small extracellular vesicles (sEVs) are potential circulating biomarkers in liquid biopsies. However, their small sizes, low abundance, and heterogeneity in molecular makeups pose major technical challenges for detecting and characterizing them quantitatively. Here, we demonstrate a single-sEV enumeration platform using lanthanide-doped upconversion nanoparticles (UCNPs). Taking advantage of the unique optical properties of UCNPs and the background-eliminating property of total internal reflection fluorescence (TIRF) imaging technique, a single-sEV assay recorded a limit of detection 1.8 × 106 EVs/mL, which was nearly 3 orders of magnitude lower than the standard enzyme-linked immunosorbent assay (ELISA). Its specificity was validated by the difference between EpCAM-positive and EpCAM-negative sEVs. The accuracy of the UCNP-based single-sEV assay was benchmarked with immunomagnetic-beads flow cytometry, showing a high correlation (R2> 0.99). The platform is suitable for evaluating the heterogeneous antigen expression of sEV and can be easily adapted for biomarker discoveries and disease diagnosis.

Quantitatively Monitoring In Situ Mitochondrial Thermal Dynamics by Upconversion Nanoparticles.

Temperature dynamics reflect the physiological conditions of cells and organisms. Mitochondria regulate the temperature dynamics in living cells as they oxidize the respiratory substrates and synthesize ATP, with heat being released as a byproduct of active metabolism. Here, we report an upconversion nanoparticle-based thermometer that allows the in situ thermal dynamics monitoring of mitochondria in living cells. We demonstrate that the upconversion nanothermometers can efficiently target mitochondria, and the temperature-responsive feature is independent of probe concentration and medium conditions. The relative sensing sensitivity of 3.2% K-1 in HeLa cells allows us to measure the mitochondrial temperature difference through the stimulations of high glucose, lipid, Ca2+ shock, and the inhibitor of oxidative phosphorylation. Moreover, cells display distinct response time and thermodynamic profiles under different stimulations, which highlight the potential applications of this thermometer to study in situ vital processes related to mitochondrial metabolism pathways and interactions between organelles.

Clinical characteristics and surgical outcomes of ependymomas in the upper cervical spinal cord: a single-center experience of 155 consecutive patients.

Ependymomas occurring in the upper cervical spinal cord (above the level of the C4 segment) are rare entities with great therapeutic challenges. This study was aimed to investigate the clinicoradiological characteristics and the prognosis in a large cohort of upper cervical ependymomas from a single institution. This retrospective study enrolled 155 patients with primary ependymomas in the upper cervical spinal cord. The pre- and post-operative clinical and magnetic resonance imaging profiles were collected. The neurological outcomes and survival events were evaluated, and potential independent risk factors were analyzed. There were 82 females and 73 males, with an average age of 43.1 ± 11.3 years. Immediately post-operatively, 118 (76.1%) patients experienced neurological deterioration and 32 (20.7%) patients remained unchanged. Three months after surgery, 61 (39.4%) patients showed deteriorated neurological functions compared to the pre-operative baseline levels. After an average follow-up period of 56.0 ± 24.7 months, the neurological functions were worse than the baseline status in 37 (23.9%) patients and improved in 33 (21.3%) patients, respectively. Logistic regression analysis identified that lower age (≤ 42 years) and lower pre-operative MMS (I-II) were independent protective factors for predicting favorable neurological functions. Multivariate Cox regression analysis revealed that incomplete resection was the only independent risk factor associated with a shorter progression-free survival. Age and pre-operative functional status affect the long-term neurological outcomes, and incomplete resection was associated with a shorter survival. Our findings indicate that gross total resection should be the goal of surgical treatment of upper cervical ependymomas.

Design and Optimization for Mounting Primary Mirror with Reduced Sensitivity to Temperature Change in an Aerial Optoelectronic Sensor.

In order to improve the image quality of the aerial optoelectronic sensor over a wide range of temperature changes, high thermal adaptability of the primary mirror as the critical components is considered. Integrated optomechanical analysis and optimization for mounting primary mirrors are carried out. The mirror surface shape error caused by uniform temperature decrease was treated as the objective function, and the fundamental frequency of the mirror assembly and the surface shape error caused by gravity parallel or vertical to the optical axis are taken as the constraints. A detailed size optimization is conducted to optimize its dimension parameters. Sensitivities of the optical system performance with respect to the size parameters are further evaluated. The configuration of the primary mirror and the flexure are obtained. The simulated optimization results show that the size parameters differently affect the optical performance and which factors are the key. The mirror surface shape error under 30 °C uniform temperature decrease effectively decreased from 26.5 nm to 11.6 nm, despite the weight of the primary mirror assembly increases by 0.3 kg. Compared to the initial design, the value of the system's modulation transfer function (0° field angle) is improved from 0.15 to 0.21. Namely, the optical performance of the camera under thermal load has been enhanced and thermal adaptability of the primary mirror has been obviously reinforced after optimization. Based on the optimized results, a prototype of the primary mirror assembly is manufactured and assembled. A ground thermal test was conducted to verify difference in imaging quality at room and low temperature, respectively. The image quality of the camera meets the requirements of the index despite degrading.

Extinction ratio and image accuracy of relayed-microgrid polarimetric imaging systems: theory and experiment.

Herein we propose a polarimetric imaging system that uses a microgrid polarizer placed on the conjugate point of two telecentric optical paths, matching large polarizers with small sensors and thus effectively decreasing optical crosstalk and increasing imaging accuracy. We define a new parameter used to construct the high-precision polarization vector transfer model under crosstalk. Using the equivalent surface of the detector, we establish the relationship between focal shift and crosstalk ratio and obtain a multi-physical coupling mathematical model that accounts for the crosstalk ratio, extinction ratio, sensor error, target vector, and imaging accuracy of the system. The relayed-microgrid polarimetric imaging system is anticipated to be able to help identify objects of interest for remote sensing and military applications.

A Median-Ratio Scene-Based Non-Uniformity Correction Method for Airborne Infrared Point Target Detection System.

Infrared detectors suffer from severe non-uniform noise which highly reduces image resolution and point target signal-to-noise ratio. This is the restriction for airborne point target detection systems in reaching the background limit. The existing methods are either not accurate enough, or too complex to be applied to engineering. To improve the precision and reduce the algorithm complexity of scene-based Non-Uniformity Correction (NUC) for an airborne point target detection system, a Median-Ratio Scene-based NUC (MRSBNUC) method is proposed. The method is based on the assumption that the median value of neighboring pixels is approximately constant. The NUC coefficients are calculated recursively by selecting the median ratio of adjacent pixels. Several experiments were designed and conducted. For both the clear sky scene and scene with clouds, the non-uniformity is effectively reduced. Furthermore, targets were detected in outfield experiments. For Target 1 48.36 km away and Target 2 50.53 km away, employing MRSBNUC the SNR of the target increased 2.09 and 1.73 times respectively compared to Two-Point NUC. It was concluded that the MRSBNUC method can reduce the non-uniformity of the detector effectively which leads to a longer detection distance and fewer false alarms of the airborne point target detection system.

Effect of sensor SNR and extinction ratio on polarimetric imaging error for nanowire-based systems.

High-sensor SNR and high extinction ratio (ER), which are often contradictory requirements for nanowire-filter-based polarimetric imaging systems, aid in attenuating polarimetric imaging system errors. Expressions were derived to analyze their attenuation effects and then simplified using photoelectronic numbers received by superpixels (PNRS). The first-derivative ratios of PNRS and ER were calculated to compare their attenuation effects. Mathematical models and experiments conducted using polarimetric imaging systems with various ERs and PNRSs indicate that systems with low PNRS and high ER exhibit a polarization error affected more by the attenuation effect of the PNRS than that of the ER. When the system ER is higher than 28, the attenuation effect of the PNRS is higher than that of the ER. Thus, system error attenuation is a trade-off between sensor SNR and ER.

Analytical method for the transformation of Zernike polynomial coefficients for scaled, rotated, and translated pupils.

Zernike polynomials provide an excellent metric basis for characterizing the wavefront aberrations of human eyes and optical systems. Since the Zernike expansion is dependent on the size, position, and orientation of the pupil in which the function is defined, it is often necessary to transform the Zernike coefficients between different pupils. An analytic method of transforming the Zernike coefficients for scaled, rotated, and translated pupils is proposed in this paper. The normalized coordinate transformation functions between the polar coordinates of the transformed pupil and the Cartesian coordinates of the original pupil are given. Based on the Cartesian and polar representations of Zernike polynomials, the coefficients' transformation matrix can be derived directly and conveniently. The first 36 terms of standard Zernike polynomials are used to validate the proposed method. For different types of transformation, transformation rules of individual Zernike terms are systematically analyzed, revealing how individual terms of the original pupil transform into terms of the transformed pupil. Numerical examples are presented to demonstrate the validity of the proposed method. Further application of the proposed method to the alignment of pupil-decentered off-axis optical systems is discussed.

Scene-based nonuniformity correction for airborne point target detection systems.

Images acquired by airborne infrared search and track (IRST) systems are often characterized by nonuniform noise. In this paper, a scene-based nonuniformity correction method for infrared focal-plane arrays (FPAs) is proposed based on the constant statistics of the received radiation ratios of adjacent pixels. The gain of each pixel is computed recursively based on the ratios between adjacent pixels, which are estimated through a median operation. Then, an elaborate mathematical model describing the error propagation, derived from random noise and the recursive calculation procedure, is established. The proposed method maintains the characteristics of traditional methods in calibrating the whole electro-optics chain, in compensating for temporal drifts, and in not preserving the radiometric accuracy of the system. Moreover, the proposed method is robust since the frame number is the only variant, and is suitable for real-time applications owing to its low computational complexity and simplicity of implementation. The experimental results, on different scenes from a proof-of-concept point target detection system with a long-wave Sofradir FPA, demonstrate the compelling performance of the proposed method.

Point target detection based on multiscale morphological filtering and an energy concentration criterion.

The research on optical imaging characteristics of infrared dim point targets in the presence of nonstationary cloud clutter and random noise is necessary for target detection. We analyze the energy concentration of point targets that are less than 3×3 pixels in size and deduce a simulation model of the point target imaging process. Then we adopt omnidirectional multiscale structural elements to detect all the possible targets distributing in every direction. The adaptive threshold and the energy concentration criterion are employed to eliminate false alarms. Finally, the trajectory of point targets is obtained after the low-order recursive correlation. The results show that the detection probability of the proposed method reaches 99.8% with 0.2% false alarm probability. It demonstrates that the proposed method has a good performance to suppress complex background and random noise. Also, it has the advantage of low complexity and easy implementation in a real-time system.

Staircase-scene-based nonuniformity correction in aerial point target detection systems.

Focal-plane arrays (FPAs) are often interfered by heavy fixed-pattern noise, which severely degrades the detection rate and increases the false alarms in airborne point target detection systems. Thus, high-precision nonuniformity correction is an essential preprocessing step. In this paper, a new nonuniformity correction method is proposed based on a staircase scene. This correction method can compensate for the nonlinear response of the detector and calibrate the entire optical system with computational efficiency and implementation simplicity. Then, a proof-of-concept point target detection system is established with a long-wave Sofradir FPA. Finally, the local standard deviation of the corrected image and the signal-to-clutter ratio of the Airy disk of a Boeing B738 are measured to evaluate the performance of the proposed nonuniformity correction method. Our experimental results demonstrate that the proposed correction method achieves high-quality corrections.

Backscanning step and stare imaging system with high frame rate and wide coverage.

Step and stare imaging with staring arrays has become the main approach to realizing wide area coverage and high resolution imagery of potential targets. In this paper, a backscanning step and stare imaging system is described. Compared with traditional step and stare imaging systems, this system features a much higher frame rate by using a small-sized array. In order to meet the staring requirements, a fast steering mirror is employed to provide backscan motion to compensate for the image motion caused by the continuously scanning of the gimbal platform. According to the working principle, the control system is designed to step/stare the line of sight at a high frame rate with a high accuracy. Then a proof-of-concept backscanning step and stare imaging system is established with a CMOS camera. Finally, the modulation transfer function of the imaging system is measured by the slanted-edge method, and a quantitative analysis is made to evaluate the performance of image motion compensation. Experimental results confirm that both high frame rate and image quality improvement can be achieved by adopting this method.

Implementation of Steiner point of fuzzy set.

This paper deals with the implementation of Steiner point of fuzzy set. Some definitions and properties of Steiner point are investigated and extended to fuzzy set. This paper focuses on establishing efficient methods to compute Steiner point of fuzzy set. Two strategies of computing Steiner point of fuzzy set are proposed. One is called linear combination of Steiner points computed by a series of crisp α-cut sets of the fuzzy set. The other is an approximate method, which is trying to find the optimal α-cut set approaching the fuzzy set. Stability analysis of Steiner point of fuzzy set is also studied. Some experiments on image processing are given, in which the two methods are applied for implementing Steiner point of fuzzy image, and both strategies show their own advantages in computing Steiner point of fuzzy set.

Cortical tension drug screen links mitotic spindle integrity to Rho pathway.

Mechanical force generation plays an essential role in many cellular functions, including mitosis. Actomyosin contractile forces mediate changes in cell shape in mitosis and are implicated in mitotic spindle integrity via cortical tension. An unbiased screen of 150 small molecules that impact actin organization and 32 anti-mitotic drugs identified two molecular targets, Rho kinase (ROCK) and tropomyosin 3.1/2 (Tpm3.1/2), whose inhibition has the greatest impact on mitotic cortical tension. The converse was found for compounds that depolymerize microtubules. Tpm3.1/2 forms a co-polymer with mitotic cortical actin filaments, and its inhibition prevents rescue of multipolar spindles induced by anti-microtubule chemotherapeutics. We examined the role of mitotic cortical tension in this rescue mechanism. Inhibition of ROCK and Tpm3.1/2 and knockdown (KD) of cortical nonmuscle myosin 2A (NM2A), all of which reduce cortical tension, inhibited rescue of multipolar mitotic spindles, further implicating cortical tension in the rescue mechanism. GEF-H1 released from microtubules by depolymerization increased cortical tension through the RhoA pathway, and its KD also inhibited rescue of multipolar mitotic spindles. We conclude that microtubule depolymerization by anti-cancer drugs induces cortical-tension-based rescue to ensure integrity of the mitotic bipolar spindle mediated via the RhoA pathway. Central to this mechanism is the dependence of NM2A on Tpm3.1/2 to produce the functional engagement of actin filaments responsible for cortical tension.

Relationship between the charge-coupled device signal-to-noise ratio and dynamic range with respect to the analog gain.

The signal-to-noise ratio and the dynamic range are the two key parameters characterizing CCD performance, especially in remote sensing applications. After exploring the possible sources of CCD noise, this paper analyzes the impacts of the analog gain on the two parameters, respectively, and establishes the mathematical models describing their relationships. Then the platforms including the CCD radiometric calibration and imaging in practice are constructed to test the proposed models based on two situations, considering the influence of the quantization noise. Finally, the design trade-off between the signal-to-noise ratio and the dynamic range is presented, such that the CCD signal-to-noise ratio will be improved as much as possible, while the dynamic range degradation becomes acceptable.

miR-340-5p Mediates Cardiomyocyte Oxidative Stress in Diabetes-Induced Cardiac Dysfunction by Targeting Mcl-1.

Diabetic cardiomyopathy (DCM) is initially characterized by early diastolic dysfunction, left ventricular remodeling, hypertrophy, and myocardial fibrosis, and it is eventually characterized by clinical heart failure. MicroRNAs (miRNAs), endogenous small noncoding RNAs, play significant roles in diabetes mellitus (DM). However, it is still largely unknown about the mechanism that links miRNAs and the development of DCM. Here, we aimed to elucidate the mechanism underlying the potential role of microRNA-340-5p in DCM in db/db mouse, which is a commonly used model of type 2 DM and diabetic complications that lead to heart failure. We first demonstrated that miR-340-5p expression was dramatically increased in heart tissues of mice and cardiomyocytes under diabetic conditions. Overexpression of miR-340-5p exacerbated DCM, which was reflected by extensive myocardial fibrosis and more serious dysfunction in db/db mice as represented by increased apoptotic cardiomyocytes, elevated ROS production, and impaired mitochondrial function. Inhibition of miR-340-5p by a tough decoy (TUD) vector was beneficial for preventing ROS production and apoptosis, thus rescuing diabetic cardiomyopathy. We identified myeloid cell leukemia 1 (Mcl-1) as a major target gene for miR-340-5p and showed that the inhibition of Mcl-1 was responsible for increased functional loss of mitochondria, oxidative stress, and cardiomyocyte apoptosis, thereby caused cardiac dysfunction in diabetic mice. In conclusion, our results showed that miR-340-5p plays a crucial role in the development of DCM and can be targeted for therapeutic intervention.

Lanthanide Ion Resonance-Driven Rayleigh Scattering of Nanoparticles for Dual-Modality Interferometric Scattering Microscopy.

Light scattering from nanoparticles is significant in nanoscale imaging, photon confinement. and biosensing. However, engineering the scattering spectrum, traditionally by modifying the geometric feature of particles, requires synthesis and fabrication with nanometre accuracy. Here it is reported that doping lanthanide ions can engineer the scattering properties of low-refractive-index nanoparticles. When the excitation wavelength matches the ion resonance frequency of lanthanide ions, the polarizability and the resulted scattering cross-section of nanoparticles are dramatically enhanced. It is demonstrated that these purposely engineered nanoparticles can be used for interferometric scattering (iSCAT) microscopy. Conceptually, a dual-modality iSCAT microscopy is further developed to identify different nanoparticle types in living HeLa cells. The work provides insight into engineering the scattering features by doping elements in nanomaterials, further inspiring exploration of the geometry-independent scattering modulation strategy.