Three-dimensional morphological and fluorescent imaging of zebrafish with a continuous-rotational light-sheet microscope

Light-sheet fluorescence microscopy(LSFM)has been widely used to image the three-dimensional(3D)structures and functions of various millimeter-size bio-specimen such as zebrafish.However,the sample adsorption and scattering cause shading of the light-sheet illumination,preventing the even 3D image of thick samples.Herein,we report a continuous-rotational light-sheet microscope(CR-LSM)that enables simultaneous 3D bright-field and fluorescence imaging.With a high-accuracy rotational stage,CR-LSM records the outline projections and the fluorescent images of the sample at multiple rotation angles.Then,3D morphology and fluorescent structure were reconstructed with a developed algorithm.Using CR-LSM,zebrafish’s whole-fish contour and blood vessel structures were obtained simultaneously.

Constructing Electron Microscope Labs: Challenges and Solutions

The construction of advanced laboratories for precision instruments, such as electron microscopes, involves unique challenges that are influenced by the specific environmental conditions required for optimal functionality. These include mitigating interference from magnetic fields and vibrations, which are critical for maintaining the precision and accuracy of the instruments used. This study aims to offer enhanced project management strategies and detailed construction solutions that address the environmental and technical needs specific to electron microscopy labs, thereby facilitating effective lab operations and extending the lifecycle of high-end precision instruments. Case studies of existing laboratory constructions, onsite investigations, and comprehensive reviews of the technical and environmental requirements provide the basis for a best practice for constructing sophisticated electron microscopy labs. The approach integrates both pre-construction planning and post-construction adjustments to create optimal operational environments. The findings suggest that successful lab constructions are those that incorporate thorough onsite assessments, strategic location choices, and the use of advanced construction materials and techniques specifically designed to counteract environmental challenges like magnetic and vibration interferences. Actionable guidelines for both planning and executing the construction of electron microscope labs highlighted in this tutorial are intended as an important resource to troubleshoot or upgrade existing lab facilities and to consult in preparation of future lab construction projects.

Deep learning method for cell count from transmitted-light microscope

Automatic cell counting provides an effective tool for medical research and diagnosis.Currently,cell counting can be completed by transmitted-light microscope,however,it requires expert knowledge and the counting accuracy which is unsatisfied for overlapped cells.Further,the image-translation-based detection method has been proposed and the potential has been shown to accomplish cell counting from transmitted-light microscope,automatically and effectively.In this work,a new deep-learning(DL)-based two-stage detection method(cGAN-YOLO)is designed to further enhance the performance of cell counting,which is achieved by combining a DL-based fluorescent image translation model and a DL-based cell detection model.The various results show that cGAN-YOLO can effectively detect and count some different types of cells from the acquired transmitted-light microscope images.Compared with the previously reported YOLO-based one-stage detection method,high recognition accuracy(RA)is achieved by the cGAN-YOLO method,with an improvement of 29.80%.Furthermore,we can also observe that cGAN-YOLO obtains an improvement of 12.11%in RA compared with the previously reported image-translation-based detection method.In a word,cGAN-YOLO makes it possible to implement cell counting directly from the experimental acquired transmitted-light microscopy images with high flexibility and performance,which extends the applicability in clinical research.

Development of a portable reflectance confocal microscope and its application in the noninvasive in vivo evaluation of mesenchymal stem cell-promoted cutaneous wound healing

The process of wound healing is routinely evaluated by histological evaluation in the clinic,which may cause scarring and secondary injury.Reflectance confocal microscopy(RCM)represents a noninvasive,real-time imaging technique that allows in vivo evaluation of the skin.Traditional RCM was wide-probe-based,which limited its application on uneven and covered skin.In this study,we report the development of a portable reflectance confocal microscope(PRCM)in which all components were assembled in a handheld shell.Although the size and weight of the PRCM were reduced based on the use of a microelectromechanical system,the resolution was kept at 0.91μm,and the field of view of the system was 343μm×532μm.When used in vivo,the PRCM was able to visualize cellular and nuclear morphology for both mouse and human skin.PRCM evaluations were then performed on wounds after topically applied mesenchymal stem cells(MSCs)or saline treatment.The PRCM allowed visualization of the formation of collagen bundles,re-epithelization from the wound edge to the wound bed,and hair follicle regeneration,which were consistent with histological evaluations.Therefore,we offer new insights into monitoring the effects of topically applied MSCs on the process of wound healing by using PRCM.This study illustrates that the newly developed PRCM represents a promising device for real-time,noninvasive monitoring of the dynamic process of wound healing,which demonstrates its potential to diagnose,monitor,or predict disease in clinical wound therapy.

Comparative Alpha Tracks Counting Using an Optical Microscope and a Spark Counter

In the metrology of radon, an environmental lung carcinogen, the integrated measurements necessary for epidemiological studies are made very often using the tracks detector LR 115 type 2. For dosimetric analysis, the etched tracks from radon alpha particles on this detector are usually counted by means of an optical microscope or a spark counter. An optimal reading of the track densities which must be converted into radon concentrations, can’t be done without a good mastery of the mode of operation and use of these devices. Furthermore, investigations to know as to whether or not each of those can be used to determine radon concentration are necessary. These are the objectives of the present work in which LR 115 samples exposed to radon for at least 3 months, were chemically developed under standard conditions and read. The track densities obtained with the microscope are very much higher than those of the counter for each sample. These results are consistent with those published by other authors. However, each of these devices can be used interchangeably for alpha tracks counting, as both provide radon concentrations with a very good linear correlation coefficient of 0.95 taking into account their respective calibration factors for the reading of this detector. In addition, the saturation phenomenon for the spark counter reading of LR 115 detector occurs beyond 11,000 tr/cm2, a density never reached during our environmental radon measurements.

Solidification process of conventional superalloy by confocal scanning laser microscope

The solidification process of a conventional superalloy, IN718, was investigated by confocal scanning laser microscope （CSLM）. The liquid fraction during solidification was obtained as a function of real time and temperature in reference with the in-situ observation. The characteristics of L→γ transformation were analyzed and the γ growing rate of each stage was also calculated. Scheil equation was employed to predict the segregation behavior, and the predict results are in consistence with the experimental results. As a result, the confocal scanning laser microscope shows a great potential for solidification process research.

Observation for Leaf Epidermal Characteristics of Four Different Pomegranate Cultivars under Scanning Electron Microscope

[Objective] The research aimed to provide reference for classification of pomegranate cultivars and studies of genetic relationship among pomegranate cultivars.[Method] The electron microscope scanning was used to comparatively observe leaf epidermal structures of 4 pomegranate cultivars.[Result] The upper epidermal structures of 4 pomegranate cultivars were similar and showed reticular structure .However, the differences existed in lower epidermis,such as cell shape,cell size and arrangement mode of cell as well as stomatal density,while the structures of leaf vein in lower epidermis of 4 pomegranate cultivars were similar.[Conclusion] The research provided morphological references for studying heterosis of pomegranate to some extent.

Obsevation for Epidermal Ultrastructure of Nostoc sphaeroides Kutzing under Scanning Electron Microscope

[Objective]The experiment aimed to explore a new way for observing surface structure of Nostoc sphaeroides Kutzing. [Method] The scanning electron microscope was used to observe the epidermal ultrastructure of wild and cultured Nostoc sphaeroides Kutzing. [ Result] The epidermis of wild and cultured Nostoc sphaeroides Kutzing showed mixture structure of fibril colloid which was reticular arranged. The difference between wild and cultured Nostoc sphaeroides Kutzing was that the outer epidermis of cultured Nostoc sphaeroides Kutzing had trichome distribution but the wild Nostoc sphaeroides Kutzing did not has such distribution. The obsevation results of under smaller than 10 μm by scanning electron microscope was touched thick and showed many folds and distortions. [ Conclusion] The scanning electron microscope was an effective way to study development of Nostoc sphaeroides Kutzing colony and it was worth popularizing.

用IM Microscope方法计算无源互调产物

在微波与射频无源互调的研究中,预测无源互调产物的电平是非常重要的。在IM Micro-scope方法的基础上进行了改进,在预测过程中直接引入输入信号经过非线性器件后输出信号的幅度和相位变化模型,给出了详细的数学模型,并实例验证了该方法的正确性。

用IM Microscope方法计算无源互调产物

在微波与射频无源互调的研究中,预测无源互调产物的电平是非常重要的,因为这样可以直观的看出无源互调噪声对于器件或者系统的影响。在众多的预测方法中,IM Microscope 方法具有很多优点。文中在IM Microscope 方法的基础上进行了改进,在预测过程中直接引入输入信号经过非线性器件后输出信号的幅度和相位变化模型,给出了详细的数学模型,这样可以大大减少很多的测量工作。

MicroScope高分辨率成像测井技术及应用

MicroScope随钻高分辨率成像测井技术是斯伦贝谢公司最新推出,能记录超高分辨率电阻率图像UHRI和多个探测深度深钮电阻率成像图,介绍了超高分辨率电阻率成像图UHRI识别岩性及地层结构、识别沉积构造、划分沉积微相、分析古水流与砂体展布等的方法,在南海西部沉积相研究方面发挥了重要作用,是值得推广的新型成像测井技术。

Observation of Helicotylenchus digonicus by Scanning Electron Microscope

[ Objective] The paper was to observe Helicotylenchus digonicus by scanning electron microscope （SEM）. [ Method ] H. digonicus collected from Changxing of Zhejiang Province was observed under scanning electron microscope, and its morphological structure was confmned under optical microscope. [ Resuit]The nematode specimens fixed by glutaraldehyde and osmium tetroxide and prepared by critical point drying were unmodified and unshrinkable with clear mor- phological structure, which could be scanned very well by scanning electron microscope. [ Conclusion ] The study provided reference for study and control of H. digonicus.

Changes of Calmodulin Distribution in the Embryo Sac of Oryza sativa Before and After Fertilization: an Immunogold Electron Microscope Study

Changes of calmodulin (CaM) distribution in the embryo sac of rice (Oryza sativa subsp. Japonica) at various stages before and after fertilization have been investigated by using immunogold electron microscopy. Before pollination, both cytoplasm and vacuoles of the egg cell, synergids and central cell were labeled by gold particles. A small amount of gold particles were localized in the nucleus, endoplasmic reticulum, mitochondria and dictyosomes. From pollination to fertilization, CaM amount increased in these cells, especially rich in the starch of amyloplasts. Increase of gold particles in the central cell began about 2 h earlier than that in the egg cell. There was no distinct difference of CaM amount between the degenerated and the persistent synergids. It is interesting to observe an obvious change of CaM distribution form during pollination and fertilization from scattered single particles to clustered particles, and back again to single particles after the fertilization finished. CaM was also localized extracellularly in the embryo sac wall as well as in the wall and intercellular space of nucellus cells. The extracellular CaM also changes in its amount and form after pollination. These results suggest that CaM, either intra- or extra-cellular, may play important roles in fertilization and zygote formation.

A Highly Stable Electrochemical Scanning Tunneling Microscope

We demonstrate a home-built electrochemical scanning tunneling microscope （ECSTM）. The ECSTM exhibits highly stable performance. The drifting rates in XY and Z directions of the ECSTM are about 67 and 55.6 pm/min, respectively. Moreover, a specially designed scanner unit successfully solves the well-known problem of large leakage current in high humidity atmosphere. The mechanical structure of the ECSTM is described in detail. The excellent performances of the system are demonstrated by the measured STM images （in copper sulfate solution）, including clean and well-ordered large area morphology of Au（111） and the atomically resolved image of highly oriented pyrolytic graphite.

Compact Scanning Tunneling Microscope with Large Scale Searching Capability for Mmultidisciplinary Applications

We present the design and performance of a home-built scanning tunneling microscope （STM）, which is compact （66 mm tall and 25 mm in diameter）, yet equipped with a 3D atomic precision piezoelectric motor in which the Z coarse approach relies on a high simplic-ity friction-type walker （of our own invention） driven by an axially cut piezoelectric tube. The walker is vertically inserted in a piezoelectric scanner tube （PST） with its brim laying at on the PST end as the inertial slider （driven by the PST） for the XZ （sample plane） motion. The STM is designed to be capable of searching rare microscopic targets （defects, dopants, boundaries, nano-devices, etc.） in a macroscopic sample area （square millimeters） under extreme conditions （low temperatures, strong magnetic elds, etc.） in which it ts. It gives good atomic resolution images after scanning a highly oriented pyrolytic graphite sample in air at room temperature.

MICRO/NANO-MACHINING ON SILICON SURFACE WITH A MODIFIED ATOMIC FORCE MICROSCOPE

To understand the deformation and removal mechanism of material on nano-scale at ultralow loads,a systemic study on AFM micro/nano-machining on single crystal ailicon is conducted. The results indicate that AFM nano- machining has a precisely dimensional controllability and a good surface quality on nanometer scale.A SEM is adopted to observe nano-machined region and chips,the results indicate that the material removal mechanisms change with the applied normal load. An XPS is used to analyze the changes of chemical composition inside and outside the nano-machined region respectively.The nano-indentation which is conducted with the same AFM diamond tip on the machined region shows a big discrepancy compared with that on the macro-scale. The calculated results show higher nano-hardness and elastic modulus than normal values .This phenomenon on be regarded as the indentation size effect(ISE).

Chalkiness Characters and Scanning Electron Microscope Observation of Rice Grain Endosperm of Japonica Varieties in Southern Henan

Chalkiness characters affect not only the grain appearance,milling,eating and cooking qualities but also the grain nutritional quality in rice,thus it is one of the most important traits in rice. It is very important for us to investigate the relation of the chalkiness formation and the development of endosperm structure and starch granule of different rice varieties. Here,we have investigated the chalkiness characters such as chalkiness rate,chalkiness degree and chalkiness area in 15 japonica rice varieties from southern Henan. Furthermore,the endosperm structure and starch granules of rice grain were also observed with scanning electron microscope. The results showed that the 15 japonica varieties have a significantly linear relationship between the chalkiness rate and chalkiness degree. Among the varieties,the biggest difference is the chalkiness rate,the second is the chalkiness area,and the last is the chalkiness degree. Moreover,there is a certain correlation between the distribution of starch granules,the arrangement of endosperm cells and the occurrence of grain chalkiness in the different rice varieties. For the same variety,the starch granules of chalky and non-chalky grains have obvious difference,while the starch granules from the transparent part of chalky rice and non-chalky rice do not have significant difference. The results would provide useful references for the improvement of grain quality in rice.

Nonlinear control techniques for an atomic force microscope system

Two nonlinear control techniques are proposed for an atomic force microscopesystem. Initially, a learning-based control algorithm is developed for the microcantilever-samplesystem that achieves asymptotic cantilever tip tracking for periodic trajectories. Specifically, thecontrol approach utilizes a learning-based feedforward term to compensate for periodic dynamics andhigh-gain terms to account for non-periodic dynamics. An adaptive control algorithm is thendeveloped to achieve asymptotic cantilever tip tracking for bounded tip trajectories despiteuncertainty throughout the system parameters. Simulation results are provided to illustrate theefficacy and performance of the control strategies.

A novel phase-sensitive scanning near-field optical microscope

Phase is one of the most important parameters of electromagnetic waves. It is the phase distribution that determines the propagation, reflection, refraction, focusing, divergence, and coupling features of light, and further affects the intensity distribution. In recent years, the designs of surface plasmon polariton （SPP） devices have mostly been based on the phase modulation and manipulation. Here we demonstrate a phase sensitive multi-parameter heterodyne scanning near-field opti- cal microscope （SNOM） with an aperture probe in the visible range, with which the near field optical phase and amplitude distributions can be simultaneously obtained. A novel architecture combining a spatial optical path and a fiber optical path is employed for stability and flexibility. Two kinds of typical nano-photonic devices are tested with the system. With the phase-sensitive SNOM, the phase and amplitude distributions of any nano-optical field and localized field generated with any SPP nano-structures and irregular phase modulation surfaces can be investigated. The phase distribution and the interference pattern will help us to gain a better understanding of how light interacts with SPP structures and how SPP waves generate, localize, convert, and propagate on an SPP surface. This will be a significant guidance on SPP nano-structure design and optimization.

Position-resolved Surface Characterization and Nanofabrication Using an Optical Microscope Combined with a Nanopipette/Quartz Tuning Fork Atomic Force Microscope

In this work, we introduce position-resolved surface characterization and nanofabrication using an optical microscope(OM) combined with a nanopipette-based quartz tuning fork atomic force microscope(nanopipette/QTF-AFM) system. This system is used to accurately determine substrate position and nanoscale phenomena under ambient conditions. Solutions consisting of 5 nm Au nanoparticles, nanowires, and polydimethylsiloxane(PDMS) are deposited onto the substrate through the nano/microaperture of a pulled pipette. Nano/microscale patterning is performed using a nanopipette/QTF-AFM, while position is resolved by monitoring the substrate with a custom OM. With this tool, one can perform surface characterization(force spectroscopy/microscopy) using the quartz tuning fork(QTF) sensor. Nanofabrication is achieved by accurately positioning target materials on the surface, and on-demand delivery and patterning of various solutions for molecular architecture.