Foot morphology and correlation with lower extremity pain in Japanese children: A cross-sectional study of the foot posture Index-6.

BACKGROUND: Abnormal foot morphology in children and adolescents is a possible risk factor for lower extremity pain. Foot posture index-6 (FPI-6) is a valid and reliable tool to assess foot morphology. However, the normative data on the age distribution for FPI-6 in Asian children and adolescents are still minimal. Further, the correlation of FPI-6 with lower extremity pain is poorly understood. We aimed to investigate the normative distribution for FPI-6 and the relationship between FPI-6 scores and knee and heel pain in Japanese children. METHODS: We included 2569 Japanese children, aged 9-15 years, at a single school from 2016 to 2018. We summarized the age distribution of children and their mean bilateral FPI-6 scores. Additionally, we assessed the tenderness at the apophysis or tendon insertions at the knee and heel. We performed a cross-sectional analysis to investigate the correlations between FPI-6 scores and sex, age, and knee and heel pain for the data obtained each year. RESULTS: The mean FPI-6 score was 3.1 ± 2.4, 3.4 ± 2.0, and 3.2 ± 1.9 for the left foot and 3.0 ± 2.4, 3.2 ± 1.9, and 3.1 ± 1.9 for the right foot in 2016, 2017, and 2018, respectively. Boys tended to have higher scores than girls, and the FPI-6 score of the left foot was significantly higher than that of the right foot (p < 0.05). There was no correlation between FPI-6 scores and knee and heel pain. CONCLUSION: Children and adolescents between 9 and 15 years of age have neutral to slightly pronated foot morphology and an average FPI-6 score of 3.0-3.4. In addition, there was no relationship between foot morphology and knee and heel pain. This normative distribution for FPI-6 in Japanese children could serve as a reference value for future research and clinical evaluation.

Do standardization of the procedure reduce measurement variability of the sonographic anterior drawer test of the ankle?

BACKGROUND: In quantitative ankle stress sonography, different examiners use different techniques, which may cause measurement variability. This study aimed to clarify whether standardizing stress sonography techniques reduces variability in the quantitative measurement of anterior talofibular ligament length change. METHODS: Fourteen examiners with a mean ultrasound experience of 8.7 years participated in this study. Each examiner performed stress ultrasonography of the ankle using their preferred method on one patient with an intact anterior talofibular ligament (Patient 1) and on two patients with chronic ankle instability (Patient 2 and 3). Changes in the ligament length between the resting and stressed positions were determined. A consensus meeting was then conducted to standardize the sonographic technique, which was used by the examiners during a repeat stress sonography on the same patients. The variance and measured values were compared between the preferred and standardized techniques using F-tests and paired t-tests, respectively. RESULTS: At a consensus meeting, a sonographic technique in which the examiner pushed the lower leg posteriorly against the fixed foot was adopted as the standardized technique. In Patient 1, the change in the anterior talofibular ligament length was 0.4 (range, -2.3-1.3) mm and 0.6 (-0.6-1.7) mm using the preferred and standardized techniques, respectively, with no significant difference in the variance (P = 0.51) or the measured value (P = 0.52). The length changes in Patient 2 were 2.0 (0.3-4.4) mm and 1.7 (-0.9-3.8) mm using the preferred and standardized techniques, respectively. In Patient 3, the length changes were 1.4 (-2.7-7.1) mm and 0.7 (-2.0-2.3) mm. There were no significant differences between the techniques in either patient group. CONCLUSION: Variability in the quantitative measurement of ankle stress sonography was not reduced despite the standardization of the technique among examiners. Hence, comparing the measured values between different examiners should be avoided.

Comparison of joint awareness after medial unicompartmental knee arthroplasty and high tibial osteotomy: a retrospective multicenter study.

INTRODUCTION: Unicompartmental knee arthroplasty (UKA) and high tibial osteotomy (HTO) are established treatments for medial compartment osteoarthritis (OA) or osteonecrosis (ON) of the knee joint, and the predominance of either procedure is inconclusive. We compared the awareness of the knee after UKA and HTO using the Forgotten joint score-12 (FJS). MATERIALS AND METHODS: This was a retrospective, multicenter study. Ninety-six knees of 90 patients who received UKA or HTO and were followed-up for at least 1 year were analyzed. Postoperative FJS was compared between the two groups and evaluated for the effect of patient-related factors and clinical outcomes. Multiple linear regression analysis was performed to predict FJS. RESULTS: There was no significant difference in the FJS between the UKA and HTO groups (p = 0.24). FJS did not correlate with any of the patient-related factors. There was a correlation between the FJS and each item of the Knee Injury and Osteoarthritis Outcome Scores (KOOS) and Lysholm Knee Scoring Scale (LKS). In multiple linear regression analysis, lower BMI, the diagnosis of OA Kellgren-Lawrence (KL) grade ≥ 3, and ON were significant predictors of better FJS. In both groups, FJS was correlated with each item of the KOOS and LKS. Internal consistency in terms of Cronbach's alpha was excellent. CONCLUSIONS: There was no significant difference in FJS between patients who underwent UKA and HTO. Lower BMI, the diagnosis of OA KL grade ≥ 3, and ON were significant predictors of better FJS.

Decreased physical activity in patients with ankle osteoarthritis. A case-control study comparing daily step counts.

BACKGROUND: The purpose of this study was to compare physical activity levels and the proportion of patients who met physical activity recommendations in patients with ankle osteoarthritis and controls. METHODS: Ankle osteoarthritis patients (n = 50) and controls (n = 50) were recruited. physical activity was measured using an accelerometer-based monitor. Physical activity parameters, including the step count/day, the proportion of patients who met physical activity recommendations of ≥7000 steps/day, and moderate to vigorous physical activity minutes/day, were compared between the patient groups. RESULTS: The patients with ankle osteoarthritis walked 3998 steps/day, while the controls walked 6531 steps/day (P < .001). Only 7 (14%) ankle osteoarthritis patients and 17 (34%) controls met the physical activity recommendations of ≥7000 steps/day (P = .01). Time spent performing moderate to vigorous physical activity was 58 and 78 min/day in the ankle osteoarthritis and controls, respectively. CONCLUSIONS: The physical activity level of ankle osteoarthritis patients was lower than that of controls. Only a limited proportion of ankle osteoarthritis patients met the physical activity recommendation.

Anatomical factors associated with progression of hallux valgus.

BACKGROUND: The purpose of this study was to clarify the incidence rate and the risk factors for the progression of the hallux valgus deformity. METHODS: Patients who had weight-bearing radiographs of the foot taken twice or more, with a ≥2-year interval, were retrospectively analyzed. Progression of the deformity was defined as an increase in the hallux valgus angle of ≥5° during the follow-up. The association of hallux valgus progression with patient characteristics and radiographic measurements at baseline was determined using univariate and multivariate analyses. RESULTS: Totally, 268 patients (217 women and 51 men; median age, 64 years) were analyzed. An increase in the hallux valgus angle of ≥5° occurred in 44 (17%) patients in a median follow-up of 49 months. Large hallux valgus angle (odds ratio, 1.07) on the dorsoplantar radiograph at baseline were independent risk factors for the progression of the deformity. CONCLUSION: Progression of the hallux valgus deformity occurred in one of six patients. Furthermore, large hallux valgus angle was the risk factor for subsequent deformity progression. Patients with large hallux valgus angle should be informed about the possible progression of the deformity.

CDKN1C-mediated growth inhibition by an EZH1/2 dual inhibitor overcomes resistance of mantle cell lymphoma to ibrutinib.

Mantle cell lymphoma (MCL) is a rare subtype of non-Hodgkin's lymphoma, which is characterized by overexpression of cyclin D1. Although novel drugs, such as ibrutinib, show promising clinical outcomes, relapsed MCL often acquires drug resistance. Therefore, alternative approaches for refractory and relapsed MCL are needed. Here, we examined whether a novel inhibitor of enhancer of zeste homologs 1 and 2 (EZH1/2), OR-S1 (a close analog of the clinical-stage compound valemetostat), had an antitumor effect on MCL cells. In an ibrutinib-resistant MCL patient-derived xenograft (PDX) mouse model, OR-S1 treatment by oral administration significantly inhibited MCL tumor growth, whereas ibrutinib did not. In vitro growth assays showed that compared with an established EZH2-specific inhibitor GSK126, OR-S1 had a marked antitumor effect on MCL cell lines. Furthermore, comprehensive gene expression analysis was performed using OR-S1-sensitive or insensitive MCL cell lines and showed that OR-S1 treatment modulated B-cell activation, differentiation, and cell cycle. In addition, we identified Cyclin Dependent Kinase Inhibitor 1C (CDKN1C, also known as p57, KIP2), which contributes to cell cycle arrest, as a direct target of EZH1/2 and showed that its expression influenced MCL cell proliferation. These results suggest that EZH1/2 may be a potential novel target for the treatment of aggressive ibrutinib-resistant MCL via CDKN1C-mediated cell cycle arrest.

Intra- and inter-observer reliability of implant positioning evaluation on a CT-based three-dimensional postoperative matching system for total knee arthroplasty.

BACKGROUND: The evaluation of postoperative total knee arthroplasty (TKA) alignment mainly relies on measurement data obtained from plain radiographs. The aim of this retrospective observational study was to document the intra- and inter-observer reliability in assessment of TKA component positioning after surgery using a three-dimensional (3D) computed tomography (CT) image matching system. METHODS: Fourteen knees from 14 patients who received primary TKA were included, and images were analyzed by blinded readers not associated with the surgeries. The examiner digitized the reference points according to defined landmarks, and the designated size component was superimposed to the 3D reconstructed CT model for measurement. In addition to the evaluation of implant position against the coronal and sagittal lower limb mechanical axes that were defined based on bony landmarks, implant position against axes connecting implant-based reference points that are easier to indicate was evaluated. RESULTS: The overall intra- and inter-observer reliabilities determined by the intraclass correlation coefficients (ICC) of the implant alignment measurement for both femoral and tibial components were good (ICC > 0.60), except in the direction of femoral flexion and extension, for both mechanical and implant-based axes. The difference between implant alignment measurements according to the traditional mechanical axis and the implant-based axis ranged between means of 0.08o and 1.70o and were statistically significantly different. CONCLUSIONS: The postoperative evaluation of implant position in the coronal and sagittal planes using 3D-CT image matching is reliable and has good reproducibility except for the sagittal alignment assessment of the femoral component. The measured implant position according to the traditional mechanical axis and the implant-based axis were slightly but significantly different.

The shape of the first metatarsal head and its association with the presence of sesamoid-metatarsal joint osteoarthritis and the pronation angle.

BACKGROUND: The purpose of this study was to evaluate the associations of the shape of the first metatarsal head with (1) the presence of osteoarthritis in the sesamoid-metatarsal joint and (2) the pronation angle of the first metatarsal head on foot radiographs. METHODS: A total of 121 patients, with the mean age of 61 years, underwent weight-bearing dorsoplantar, lateral, and first metatarsal axial radiographs. The shape of the first metatarsal head's lateral edge was classified as either rounded, intermediate, or angular in shape in the dorsoplantar view. The presence of osteoarthritis in the sesamoid-metatarsal joint and the pronation angle of the first metatarsal head were assessed in the first metatarsal axial view. Other variables that could affect the first metatarsal shape, including the lateral first metatarsal inclination angle, were also assessed. Univariate and multivariate analyses were performed to determine the associations. RESULTS: The prevalence of sesamoid-metatarsal osteoarthritis was significantly higher (77%, 27%, and 29% for rounded, intermediate, and angular, respectively, P < .001), and the metatarsal pronation angle was significantly larger (14°, 8°, and 4° for rounded, intermediate, and angular, respectively, P < .001) in feet with a rounded metatarsal head. These associations were also significant in the multiple regression analysis. CONCLUSION: A rounded metatarsal head was associated with a higher prevalence of osteoarthritis within the sesamoid-metatarsal joint, as well as a larger first metatarsal head pronation angle. A negative round sign can be used as a simple indicator of an effective correction to the first metatarsal pronation angle during hallux valgus surgery. However, in feet with sesamoid-metatarsal osteoarthritis, surgeons will need to be cautious as overcorrection may occur.

Influence of foot position on the measurement of first metatarsal axial rotation using the first metatarsal axial radiographs.

BACKGROUND: The purpose of this study was to clarify 1) the measurement error of the pronation angle using the first metatarsal axial radiograph with the pronation angle along the longitudinal axis of the first metatarsal as the reference standard, 2) the influence of variability in the foot position on the measurement error, and 3) the intra- and interrater reliability of pronation angle measurement using digitally reconstructed radiographs. METHODS: Digitally reconstructed radiographs of the first metatarsal were generated from the computed tomography images of 10 feet without hallux valgus (non-HV group) and 10 feet with hallux valgus (HV group). In total, 135 images were created at different degrees of supination, plantarflexion, and adduction from each foot to simulate the first metatarsal axial view. Then, the pronation angle of the first metatarsal was measured. The measurement error was determined using the mean error and 95% limits of agreement. Simple linear regression analysis was used to test the correlations of the measurement error with pronation, plantarflexion and adduction angles. The intra- and interrater reliability of measurement was assessed using the intraclass correlation coefficient and minimum detectable change values. RESULTS: The mean measurement errors were 0.1° for both the non-HV and HV groups. There was no significant correlation of the measurement error with pronation, plantarflexion or adduction angles for both groups. Additionally, the intraclass correlation coefficients for the intra- and interrater reliability were more than 0.9 in both the non-HV and HV groups with the minimum detectable change values ranging from 0.7° to 1.4°. CONCLUSION: The measurement error of first metatarsal pronation using the axial view was clinically acceptable. The measurements were not influenced by the variability in foot position while obtaining the radiograph. The first metatarsal axial view could be used to quantify the first metatarsal coronal rotation.

Dual inhibition of enhancer of zeste homolog 1/2 overactivates WNT signaling to deplete cancer stem cells in multiple myeloma.

Multiple myeloma (MM) is an incurable hematological malignancy caused by accumulation of abnormal clonal plasma cells. Despite the recent development of novel therapies, relapse of MM eventually occurs as a result of a remaining population of drug-resistant myeloma stem cells. Side population (SP) cells show cancer stem cell-like characteristics in MM; thus, targeting these cells is a promising strategy to completely cure this malignancy. Herein, we showed that SP cells expressed higher levels of enhancer of zeste homolog (EZH) 1 and EZH2, which encode the catalytic subunits of Polycomb repressive complex 2 (PRC2), than non-SP cells, suggesting that EZH1 as well as EZH2 contributes to the stemness maintenance of the MM cells and that targeting both EZH1/2 is potentially a significant therapeutic approach for eradicating myeloma stem cells. A novel orally bioavailable EZH1/2 dual inhibitor, OR-S1, effectively eradicated SP cells and had a greater antitumor effect than a selective EZH2 inhibitor in vitro and in vivo, including a unique patient-derived xenograft model. Moreover, long-term continuous dosing of OR-S1 completely cured mice bearing orthotopic xenografts. Additionally, PRC2 directly regulated WNT signaling in MM, and overactivation of this signaling induced by dual inhibition of EZH1/2 eradicated myeloma stem cells and negatively affected tumorigenesis, suggesting that repression of WNT signaling by PRC2 plays an important role in stemness maintenance of MM cells. Our results show the role of EZH1/2 in the maintenance of myeloma stem cells and provide a preclinical rationale for therapeutic application of OR-S1, leading to significant advances in the treatment of MM.

Investigating the Potential of Multilineage Differentiating Stress-Enduring Cells for Osteochondral Healing.

OBJECTIVE: Multilineage differentiating stress-enduring (Muse) cells, a pluripotent stem cell subset of mesenchymal stem cells (MSCs), have shown promise for various tissue repairs due to their stress tolerance and multipotent capabilities. We aimed to investigate the differentiation potential in vitro, the dynamics in vivo, and the reparative contribution of Muse cells to osteochondral lesions. DESIGN: Labeled MSCs were cultured and sorted into Muse and non-Muse (MSCs without Muse cells) groups. These cells were then formed into spheroids, and chondrogenic differentiation was assessed in vitro. Twenty-one immunocompromised mice were used as the in vivo models of osteochondral lesions. Live imaging, macroscopic evaluation, and histological and immunohistochemical analyses were conducted at the 4- and 8-week time points. RESULTS: Muse cell spheroids were formed, which were larger and stained more intensely with toluidine blue than non-Muse spheroids, indicating better chondrogenic differentiation. Live imaging confirmed luminescence in all 4-week model knees, but only in a few knees at 8 weeks, suggesting cell persistence. Macroscopically and histologically, no significant differences were observed between the Muse and non-Muse groups at 4 and 8 weeks; however, both groups showed better cartilage repair than that of the vehicle group at 8 weeks. No collagen type II generation was observed in the repaired tissues. CONCLUSION: The implantation of the spheroids of Muse and non-Muse cells resulted in better healing of osteochondral lesions than that of the controls, and Muse cells had a higher chondrogenic differentiation potential in vitro than non-Muse cells.

Deep learning generates custom-made logistic regression models for explaining how breast cancer subtypes are classified.

Differentiating the intrinsic subtypes of breast cancer is crucial for deciding the best treatment strategy. Deep learning can predict the subtypes from genetic information more accurately than conventional statistical methods, but to date, deep learning has not been directly utilized to examine which genes are associated with which subtypes. To clarify the mechanisms embedded in the intrinsic subtypes, we developed an explainable deep learning model called a point-wise linear (PWL) model that generates a custom-made logistic regression for each patient. Logistic regression, which is familiar to both physicians and medical informatics researchers, allows us to analyze the importance of the feature variables, and the PWL model harnesses these practical abilities of logistic regression. In this study, we show that analyzing breast cancer subtypes is clinically beneficial for patients and one of the best ways to validate the capability of the PWL model. First, we trained the PWL model with RNA-seq data to predict PAM50 intrinsic subtypes and applied it to the 41/50 genes of PAM50 through the subtype prediction task. Second, we developed a deep enrichment analysis method to reveal the relationships between the PAM50 subtypes and the copy numbers of breast cancer. Our findings showed that the PWL model utilized genes relevant to the cell cycle-related pathways. These preliminary successes in breast cancer subtype analysis demonstrate the potential of our analysis strategy to clarify the mechanisms underlying breast cancer and improve overall clinical outcomes.

Changes in the Syndesmotic Reduction After Syndesmotic Suture-Button Fixation for Ankle Malleolar Fractures: 1-Year Longitudinal Evaluations Using Computed Tomography.

BACKGROUND: Evaluation over time is important in assessing the reduction of the syndesmosis after suture-button fixation for ankle malleolar fractures. The purposes of this study were to evaluate time-dependent change in the syndesmotic reduction immediately after suture-button fixation for ankle malleolus fractures and 1 year after surgery using computed tomography, and to investigate the reliability of the measurement values to evaluate the reduction of syndesmosis. METHODS: We assessed 28 patients who underwent suture-button fixation for ankle fractures. Syndesmotic reduction was assessed within 2 weeks of the fracture surgery and 1 year after surgery using axial computer tomographic images. Side-to-side differences in the anterior, central, and posterior tibiofibular distances, anteroposterior fibular translation, fibular rotation, and syndesmosis area were measured. RESULTS: The mean anterior tibiofibular distance and anteroposterior fibular translation were 1.8 mm and 1.5 mm, respectively, after syndesmotic fixation. They decreased to 1.2 mm and 0.6 mm, respectively, at 1 year after surgery (P = .03 and P = .01, respectively). The other measurement values did not change over time. The minimum detectable change in the distance of measurements was 1 mm or less. CONCLUSION: The anterior tibiofibular distance and anteroposterior fibular translation had decreased 1 year after fixation in ankle malleolar fractures with syndesmotic suture button. Even if the fibula is posteriorly malreduced by the time computed tomography is performed immediately after surgery, the fibula may return to a good position 1 year after surgery. LEVEL OF EVIDENCE: Level IV, case series.

Randomized comparative study of suspension femoral fixation device in graft position maintenance in anterior cruciate ligament reconstruction: EndoButton CL vs TightRope RT.

BACKGROUND: In double-bundle anterior cruciate ligament reconstruction (ACLR), fixed-loop and adjustable-loop cortical suspensory devices are commonly used to fix the soft graft on the femoral side. However, few studies have compared in vivo elongation of the two devices. The purpose of this study was to determine whether EndoButton CL (EB) and TightRope RT (TR), the suspensory fixation devices used in ACLR, maintained their length in vivo from the time of surgery through the postoperative period in a randomized controlled trial. METHODS: This study prospectively incorporated 30 patients undergoing initial ACLR at a single center. Participants were divided into two groups using a stratified randomization method with age and sex as assignment adjustment factors. EB or TR was used for fixation of the soft graft on the femoral side. The primary endpoint was to compare the elongation distance of the suspensory device. MRIs were taken within seven days after ACLR and 3,6,12 months postoperatively and measured by a radiologist in a blinded fashion. Secondary endpoints included the side-to-side difference in anterior translation, one leg hop test (HOP index), Lachman test, lateral pivot shift test, and Lysholm score one year postoperatively. RESULTS: Twenty-eight patients (EB, n = 13; TR, n = 15) were followed for one year. There was no significant difference between EB and TR groups in elongation from the immediate postoperative period to 3, 6, 12 months after surgery. However, the non-inferiority of TR to EB (non-inferiority margin: 1.5 mm) was not proved by the difference in measured elongation between the two groups (TR - EB, lower 95% CI. AM: 1.80 mm; PL: 1.86 mm) at 6 months. There was no significant difference in anterior translation, HOP index, Lachman test, lateral pivot shift test, or Lysholm score. CONCLUSION: EB and TR had similar graft retaining ability in vivo for 12 months, but the non-inferiority of TR against EB was not verified statistically.

Effect of Systemic Administration of Granulocyte Colony-Stimulating Factor on a Chronic Partial-Thickness Cartilage Defect in a Rabbit Knee Joint.

OBJECTIVE: Cartilage lesions in the knee joint can lead to joint mechanics changes and cause knee pain. Bone marrow stimulation (BMS) promotes cartilage regeneration by perforating the subchondral bone just below the injury and inducing bone marrow cells. This study aimed to investigate whether systemic administration of granulocyte colony-stimulating factor (G-CSF) with BMS improves repair of chronic partial-thickness cartilage defects (PTCDs). DESIGN: Eighteen 6-month-old New Zealand white rabbits were divided into 3 groups: control (C, n = 6), BMS alone (n = 6), and BMS + G-CSF (n = 6). Partial cartilage defects with 5 mm diameter were created in the trochlear region of both knees; after 4 weeks, the BMS alone and BMS + G-CSF groups underwent BMS; G-CSF (50 µg/kg) or saline was administered subcutaneously for 5 days starting from 3 days before BMS. At 8 and 16 weeks after cartilage defect creation, the area of cartilage defects was macroscopically and histologically evaluated. RESULTS: International Cartilage Repair Society (ICRS) grades for macroscopic assessment were 0, 0.7, and 0.7 at 8 weeks and 0, 1.2, and 1.3 at 16 weeks in the C, BMS, and BMS + G-CSF groups, respectively. Wakitani scores for histological assessment were 9.8, 8.7, and 8.2 at 8 weeks and 9.5, 9, and 8.2 at 16 weeks in the C, BMS, and BMS + G-CSF groups, respectively. The BMS + G-CSF group showed significantly more repair than the C group, but there was no difference from the BMS group. CONCLUSIONS: The effect of BMS and G-CSF on chronic PTCDs in mature rabbit knees was limited.

Lensless fourier transform electron holography applied to vortex beam analysis.

Lensless Fourier transform holography has been developed. By treating Bragg diffraction waves as object waves and a transmitted spherical wave as a reference wave, these two waves are interfered and recorded as holograms away from the reciprocal plane. In this method, reconstruction of holograms requires only one Fourier transform. Application of this method to analyze vortex beams worked well and their amplitude and phase distributions were obtained on the reciprocal plane. By combining the conventional holography with the developed lensless Fourier transform holography, we can reconstruct and analyze electron waves from the real to reciprocal space continuously.

Electron holography on Fraunhofer diffraction.

Electron holography in Fraunhofer region was realized by using an asymmetric double slit. A Fraunhofer diffraction wave from a wider slit worked as an objective wave interfered with a plane wave from a narrower slit as a reference wave under the pre-Fraunhofer condition and recorded as a hologram. Here, the pre-Fraunhofer condition means that the following conditions are simultaneously satisfied: single-slit observations are performed under the Fraunhofer condition and the double-slit observations are performed under the Fresnel condition. Amplitude and phase distributions of the Fraunhofer diffraction wave were reconstructed from the hologram by the Fourier transform reconstruction method. The reconstructed amplitude and phase images corresponded to Fraunhofer diffraction patterns; in particular, the phase steps of π at each band pattern in the phase image were confirmed. We hope that the developed Fraunhofer electron holography can be extended to a direct phase detection method in the reciprocal space.

Interference experiment with asymmetric double slit by using 1.2-MV field emission transmission electron microscope.

Advanced electron microscopy technologies have made it possible to perform precise double-slit interference experiments. We used a 1.2-MV field emission electron microscope providing coherent electron waves and a direct detection camera system enabling single-electron detections at a sub-second exposure time. We developed a method to perform the interference experiment by using an asymmetric double-slit fabricated by a focused ion beam instrument and by operating the microscope under a "pre-Fraunhofer" condition, different from the Fraunhofer condition of conventional double-slit experiments. Here, pre-Fraunhofer condition means that each single-slit observation was performed under the Fraunhofer condition, while the double-slit observations were performed under the Fresnel condition. The interference experiments with each single slit and with the asymmetric double slit were carried out under two different electron dose conditions: high-dose for calculation of electron probability distribution and low-dose for each single electron distribution. Finally, we exemplified the distribution of single electrons by color-coding according to the above three types of experiments as a composite image.

Illumination semiangle of 10-9 rad achieved in a 1.2-MV atomic resolution holography transmission electron microscope.

The coherency of a 1.2-MV transmission electron microscope was evaluated through illumination semiangles calculated from lengths over which Fresnel fringes can be observed. These lengths were determined from the diameters of circular holes fully filled with Fresnel fringes, i.e. this method allows lengths to be accurately measured even if micrographs are subjected to distortions. The smallest illumination semiangle of 4.0 × 10-9 rad was obtained for a circular hole with a diameter of 191 µm. In addition, electron beam brightness was estimated to be approximately 3 × 1014 A/m2·sr from the obtained illumination semiangle values and current densities. The results provide us with essential information that can be referred to in future electron holography studies aimed at detecting weak electromagnetic fields in materials.