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.

Investigation of bubbles escape behavior from low basicity mold flux for high-Mn high-Al steels using 3D X-ray microscope

During the continuous casting process of high-Mn high-Al steels,various types of gases such as Ar need to escape through the top of the mold.In which,the behavior of bubbles traversing the liquid slag serves as a restrictive link,closely associated with viscosity and the thickness of liquid slag.In contrast to two-dimensional surface observation,three-dimensional(3D)analysis method can offer a more intuitive,accurate,and comprehensive information.Therefore,this study employs a 3D X-ray microscope(3D-XRM)to obtained spatial distribution and 3D morphological characteristics of residual bubbles in mold flux under different basicity of liquid slag,different temperatures,and different holding times.The results indicate that as basicity of slag increases from 0.52 to 1.03,temperature increases from 1423 to 1573 K,the viscosity of slag decreases,the floating rate of bubbles increases.In addition,when holding time increases from 10 to 30 s,the bubbles floating distance increases,and the volume fraction and average equivalent sphere diameter of the bubbles solidified in the mold flux gradually decreases.In one word,increasing the basicity,temperature,and holding time leading to an increase in the removal rate of bubbles especially for the large.These findings of bubbles escape behavior provide valuable insights into optimizing low basicity mold flux for high-Mn high-Al steels.

Micro-Scanning Error Correction Technique for an Optical Micro-Scanning Thermal Microscope Imaging System

An error correction technique for the micro-scanning instrument of the optical micro-scanning thermal microscope imaging system is proposed. The technique is based on micro-scanning technology combined with the proposed second-order oversampling reconstruction algorithm and local gradient image reconstruction algorithm. In this paper, we describe the local gradient image reconstruction model, the error correction technique, down-sampling model and the error correction principle. In this paper, we use a Lena original image and four low-resolution images obtained from the standard half-pixel displacement to simulate and verify the effectiveness of the proposed technique. In order to verify the effectiveness of the proposed technique, two groups of low-resolution thermal microscope images are collected by the actual thermal microscope imaging system for experimental study. Simulations and experiments show that the proposed technique can reduce the optical micro-scanning errors, improve the imaging effect of the system and improve the system's spatial resolution. It can be applied to other electro-optical imaging systems to improve their resolution.

Research on a Diamond Tip Wear Mechanism in Atomic Force Microscope-based Micro/nano-machining

The object is to investigate the wear of an atomic force microscope (AFM) diamond tip when conducting micro/nano machining on single crystal silicon surface. The experimental research and theoretical analysis were carried out on the worn tip in terms of wear rate, wear mechanism and the effect of the tip wear on micro machining process. The wear rate was calculated as 1.7(10～10mm 3/(N·m) by using a theoretical model combined with the experimental results. Through an integration of an AFM observation on the worn tip features with the FEM simulation of the stress distribution, in addition to the unit cutting force calculation on the AFM diamond tip, the wear mechanism of the AFM diamond tip was concluded as mainly chemical wear, and the wear process was also elaborated as well.

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.

PROBE-HOLE FIELD EMISSION MICROSCOPE SYSTEM CONTROLLED BY COMPUTER

A probe-hole field emission microscope system,controlled by the Apple Ⅱ computer,has been developed and operated successfully for measuring the work function of a single crystalplane.The detection screen,the phototube and the amplifier are combined for measuring theprobe-hole current.The combination is calibrated and the calibrated data are used in the com-puter program.The high voltage on the viewing screen is adjusted by using a D/A converter.The total current and the probe-hole current are acquired by using an A/D converter.A programin BASIC is used for processing all data and the Fowler-Nordheim plot parameters are given.Work functions of single crystal planes can then be calculated;as examples,the work functionson the clean W(100)and W(111)planes are measured to be 4.67 eV and 4.45 eV,respectively.

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.

Constant Force Feedback Controller Design Using PID-Like Fuzzy Technique for Tapping Mode Atomic Force Microscopes

A novel constant force feedback mechanism based on fuzzy logic for tapping mode Atomic Force Microscopes (AFM) is proposed in this paper. A mathematical model for characterizing the cantilever-sample interaction subsystem which is nonlinear and contains large uncertainty is first developed. Then, a PID-like fuzzy controller, combing a PD-like fuzzy controller and a PI controller, is designed to regulate the controller efforts and schedule the applied voltage of the Z-axis of the piezoelectric tube scanner to maintain a constant tip-sample interaction force during sample-scanning. Using the PID-like fuzzy controller allows the cantilever tip to track sample surface rapidly and accurately even though the topography of the surface is arbitrary and not given in advance. This rapid tracking response facilitates us to observe samples with high aspect ratio micro structures accurately and quickly. Besides, the overshoot which will result in tip crash in commercial AFMs with a traditional PID controller could be avoided. Additionally, the controller efforts can be intelligently scheduled by using the fuzzy logic. Thus, continuous manual gain-tuning by trial and error such as those in commercial AFMs is alleviated. In final, computer simulations and experimental verifications are provided to demonstrate the effectiveness and confirm the validity of the proposed controller.

A stage-scanning laser confocal microscope and protocol for DNA methylation sequencing

Recent understanding of the role of epigenetic regulation in health and disease has necessitated the development of newer and efficient methods to map the methylation pattern of target gene. In this article we report construction of a stage-scanning laser confocal microscope (SLCM) and associated protocol that determines the methylation status of target gene. We have adapted restricted Sanger’s sequencing where fluorescine labeled primers and dideoxy guanine fraction alone are used for target amplification and termination at cytosine positions. Amplified ssDNA bands are separated in 6% denaturing PAGE and scanned using SLCM to sequence the positions of methylated cytosines. We demonstrate that our me- thodology can detect < 100 femtomoles of DNA, and resolve the position of cytosine within ± 2 nucleotide. In a calibration run using a designer DNA of 99 bases, our methodology had resolved all the 11 cytosine positions of the DNA. We have further demonstrated the utility of apparatus by mapping methylation status in the Exon-1 region of a gene, E-Cadherin, in the plasma DNA sample of a healthy subject. We believe our approach constitute a low cost alternative to conventional DNA sequencers and can help develop methylation based DNA biomarkers for the diagnosis of disease and in therapeutics.

EFFECT OF ACUPUNCTURE ANALGESIA ON SYNAPSFS OF NUCLEUS CENTRO—MEDIANUS OBSERVED WITH ELECTRO—MICROSCOPE

The present study was aimed at the investigation of the ultrastructuralchanges of nucleus centromedianus(CM)of nuclei intralaminares under acupuncture analge-sia.The results of the observations on 18 healthy and adult rats were as follows.1.Thesynapses of the CM belong mainly to the axodendricular synapses and also with some suc-cessive or juxtaposing synapses.2.The presynaptic sac contains a lot of round and clearvesicles,some granular vesicles also spread among clear vesicles in a few synapses.3.Theround clear synaptic vesicles of CM in the electroacupuncture analgesia(EA)group are de-creased significantly than that in the control group.It is suggested that CM plays a role inacupuncture analgesia.

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.

ELECTRON MICROSCOPE ANALYSIS OF MITOCHONDRIA ON RENAL TUBULE CELL SUFFERED WARM ISCHEMIC AND REPER FUSIVE DAMAGE IN RABBITS

ＥＬＥＣＴＲＯＮＭＩＣＲＯＳＣＯＰＥＡＮＡＬＹＳＩＳＯＦＭＩＴＯＣＨＯＮＤＲＩＡＯＮＲＥＮＡＬＴＵＢＵＬＥＣＥＬＬＳＵＦＦＥＲＥＤＷＡＲＭＩＳＣＨＥＭＩＣＡＮＤＲＥＰＥＲＦＵＳＩＶＥＤＡＭＡＧＥＩＮＲＡＢＢＩＴＳＥＬＥＣＴＲＯＮＭＩＣＲＯＳＣＯＰＥＡ...

Absolute Internal Energy of the Real Gas

The internal energy U of the real, neutral-gas particles of total mass M in the volume V can have positive and negative values, whose regions are identified in the state chart of the gas. Depending on the relations among gas temperature T, pressure p and mass-specific volume v=V/M, the mass exists as a uniform gas of freely-moving particles having positive values U or as more or less structured matter with negative values U. In the regions U>0?above the critical point [Tc , pc , vc] it holds that p(T,v)>pc and v>vc, and below the critical point it holds that p(T,v)c and v>vv , where vv is the mass-specific volume of saturated vapor. In the adjacent regions with negative internal energy values Uc is the line of equal positive and negative energy contributions and thus represents a line of vanishing internal energy ?U=0. At this level along the critical isochor the ever present microscopic fluctuations in energy and density become macroscopic fluctuations as the pressure decreases on approaching the critical point;these are to be observed in experiments on the critical opalescence. Crossing the isochor vc from U>0 to UΔU>0 happens without any discontinuity. The saturation line vv also separates the regions between U>0 and U , but does not represent a line U=0. The internal-energy values of saturated vapor Uv and condensate Ui can be determined absolutely as functions of vapor pressure p and densities (M/V)v and (M/V)i , repectively, yielding the results Uiv, U=Ui+Uvc and U=Ui=Uv=0 at the critical point. Crossing the line Vv from U=Uv>0 to U=Uv+UiΔU=-Ui>0 to be removed from the particle system. The thermodynamic and quantum-mechanical formulations of the internal energy of a particle system only agree if both the macroscopic and microscopic energy scales have the same absolute energy reference value 0. Arguments for the energy reference value in the state of transition from bound to freely- moving particles in macroscopic classical and microscopic quantum particle systems are discussed.

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.

STUDY ON THE SCANNING PROBE MICROSCOPE WITH WIDE MEASURING RANGE

ＳＴＵＤＹＯＮＴＨＥＳＣＡＮＮＩＮＧＰＲＯＢＥＭＩＣＲＯＳＣＯＰＥＷＩＴＨＷＩＤＥＭＥＡＳＵＲＩＮＧＲＡＮＧＥＺｈａｎｇＨｏｎｇｈａｉ，ＣａｏＷｅｉ，ＬｉＷｅｎｊｕ，ＳｈｉＨａｎｍｉｎ，ＣｈｅｎＲｉｙａｏＨｕａｚｈｎｇＵｎｉｖｅｒｓｉｔｙｏｆＳｃｉｅ...

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.

Thermal Diffusion of Si Atoms at the Interface of Mo/Si Bilayers Studied with a Soft X-ray Emission Microscope

Thermal diffusion of Si atoms at the interface in Mo/Si multilayers was observed with an imaging type soft X ray emission microscope developed by us. It was possible to observe the diffusion with 0.2nm depth resolution in the direction normal to the interface by comparing the emission intensity for exactly the same position. The diffusion coefficient of Si atoms in Mo at 600℃ was roughly estimated to be 6.0×10 17 cm 2/s.

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.

Qualitative analysis of re mineralized carious lesions subjected to fluoride supplement through confocal laser scanning microscope

Aim: 1] Comparative evaluation of the linear depth of induced remineralized lesions after subjecting to fluoride supplements and 2] To assess the average fluorescence at both the demineralized and the remi-neralized zones in all the three study groups under confocal laser scanning microscope. Method: Forty five sound human premolars extracted for orthodon-tic reasons were decoronated 1 mm below the ce-mento-enamel junction and coated with nail varnish except for a 3 × 3 mm window on the buccal surface. The samples were placed in 50 ml of de mineralizing solution at pH 4.6 for 96 hours. Following deminera-lization, the lower half of the 3 × 3 mm window in all the samples were covered with nail varnish to serve as control. The samples were randomly divided into three groups of fifteen teeth each (n = 15) and speci-mens in group A[Nfd] were remineralized using non-fluoridated dentifrice [control], those in groups B [Fd5] and group C [Fd10] using 500 ppm and 1000 ppm of fluoride containing dentifrice, respectively. The specimens were subjected to a 20 day reminera-lization treatment regimen and were sectioned into 100 μm thick sections and two images were captured on the buccal surface from either side of the midpoint of occluso-cervical length using confocal laser scan-ning microscope [CLSM]. Results: were tabulated and statistically analyzed by Anova. Study concluded that 1000 ppm fluoridated dentifrice showed a greater degree of remineralization than other groups and confocal laser scanning microscopes gives promising results in the diagnosis of early enamel lesions over the conventional methods.

New Method for Injectable Quinine Quality Assurance Control Using a Multi-Spectral Microscope

Africa is the world region that is most affected by malaria. Among the therapies used, injectable quinine is considered to be one of the effective antimalarial drug, however non-quality assured antimalarials clearly have a strong market penetration across Africa. To overcome this problem, it becomes more and more necessary to set up quantitative and qualitative analysis system for antimalarial quality control. The objective of the present investigation is an attempt to use customized multispectral microscope equipped with UV-Visible lasers for injectable quinine quality assurance control routinely. For that, we have established the calibration curve of quinine solution concentration as a function of area under light intensity histogram crossing the solution. From this calibration curve, we can check the conformity of any injectable quinine according to the pharmacopoeia involved. The proposed technique is a promising alternative for drug quality control routinely.