The GA-BPNN-Based Evaluation of Cultivated Land Quality in the PSR Framework Using Gaofen-1 Satellite Data.

Rapid and efficient assessment of cultivated land quality (CLQ) using remote sensing technology is of great significance for protecting cultivated land. However, it is difficult to obtain accurate CLQ estimates using the current satellite-driven approaches in the pressure-state-response (PSR) framework, owing to the limitations of linear models and CLQ spectral indices. In order to improve the estimation accuracy of CLQ, this study used four evaluation models (the traditional linear model; partial least squares regression, PLSR; back propagation neural network, BPNN; and BPNN with genetic algorithm optimization, GA-BPNN) to evaluate CLQ for determining the accurate evaluation model. In addition, the optimal satellite-derived indicator in the land state index was selected among five vegetation indices (the normalized vegetation index, NDVI; enhanced vegetation index, EVI; modified soil-adjusted vegetation index, MSAVI; perpendicular vegetation index, PVI; and soil-adjusted vegetation index, SAVI) to improve the prediction accuracy of CLQ. This study was conducted in Conghua District of Guangzhou, Guangdong Province, China, based on Gaofen-1 (GF-1) data. The prediction accuracies from the traditional linear model, PLSR, BPNN, and GA-BPNN were compared using observations. The results demonstrated that (1) compared with other models (the traditional linear model: R2 = 0.14 and RMSE = 91.53; PLSR: R2 = 0.33 and RMSE = 74.58; BPNN: R2 = 0.50 and RMSE = 61.75), the GA-BPNN model based on EVI in the land state index provided the most accurate estimates of CLQ, with the R2 of 0.59 and root mean square error (RMSE) of 56.87, indicating a nonlinear relationship between CLQ and the prediction indicator; and (2) the GA-BPNN-based evaluation approach of CLQ in the PSR framework was driven to map CLQ of the study area using the GF-1 data, leading to an RMSE of 61.44 at the regional scale, implying that the GA-BPNN-based evaluation approach has the potential to map CLQ over large areas. This study provides an important reference for the high-accuracy prediction of CLQ based on remote sensing technology.

The Optimal Image Date Selection for Evaluating Cultivated Land Quality Based on Gaofen-1 Images.

This study proposes a method for determining the optimal image date to improve the evaluation of cultivated land quality (CLQ). Five vegetation indices: leaf area index (LAI), difference vegetation index (DVI), enhanced vegetation index (EVI), normalized difference vegetation index (NDVI), and ratio vegetation index (RVI) are first retrieved using the PROSAIL model and Gaofen-1 (GF-1) images. The indices are then introduced into four regression models at different growth stages for assessing CLQ. The optimal image date of CLQ evaluation is finally determined according to the root mean square error (RMSE). This method is tested and validated in a rice growth area of Southern China based on 115 sample plots and five GF-1 images acquired at the tillering, jointing, booting, heading to flowering, and milk ripe and maturity stage of rice in 2015, respectively. The results show that the RMSEs between the measured and estimated CLQ from four vegetation index-based regression models at the heading to flowering stage are smaller than those at the other growth stages, indicating that the image date corresponding with the heading to flowering stage is optimal for CLQ evaluation. Compared with other vegetation index-based models, the LAI-based logarithm model provides the most accurate estimates of CLQ. The optimal model is also driven using the GF-1 image at the heading to flowering stage to map CLQ of the study area, leading to a relative RMSE of 14.09% at the regional scale. This further implies that the heading to flowering stage is the optimal image time for evaluating CLQ. This study is the first effort to provide an applicable method of selecting the optimal image date to improve the estimation of CLQ and thus advanced the literature in this field.

Microarray Expression Profile of Circular RNAs in Peripheral Blood Mononuclear Cells from Active Tuberculosis Patients.

BACKGROUND/AIMS: Dysregulated expression of circular RNAs (circRNAs) was demonstrated to be implicated in many diseases. Here, we aimed to determine circRNA profile in peripheral blood mononuclear cells (PBMCs) from active tuberculosis (TB) patients to identify novel biomarkers for TB. METHODS: Expression profile of circRNAs in PBMCs from 3 active pulmonary TB patients and 3 healthy controls were analyzed by microarray assay. Six circRNAs were selected for validation using real time-quantitative PCR (qRT-PCR) in 40 TB patients and 40 control subjects. Receiver operating characteristic (ROC) curve was constructed to evaluate their values in TB diagnosis. Hsa_circRNA_001937 was chosen for further evaluation in an independent cohort consisting of 115 TB, 40 pneumonia, 40 COPD, 40 lung cancer patients and 90 control subjects. An eight-month follow up was performed in 20 newly diagnosed TB patients to investigate the expression change of hsa_circRNA_001937 after chemotherapy. RESULTS: We revealed and confirmed that a number of circRNAs were dysregulated in TB patients. Of the six studied physio circRNAs, the levels of hsa_circRNA_001937, hsa_circRNA_009024 and hsa_ circRNA_005086 were significantly elevated and hsa_circRNA_102101, hsa_circRNA_104964 and hsa_circRNA_104296 were significantly reduced in PBMCs from TB patients as compared to healthy controls. ROC curve analysis suggested that hsa_circRNA_001937 has the largest area under the curve (AUC = 0.873, P<0.001). Hsa_circRNA_001937 was significantly increased in patients with TB compared with patients with pneumonia, COPD and lung cancer. Hsa_ circRNA_001937 was correlated with TB severity (r = 0.4053, P = 0.010) and its expression significantly decreased after treatment. CONCLUSION: This study identified a set of deregulated circRNAs in active TB PBMCs, our data also suggest that hsa_circRNA_001937 can be used as a potential diagnostic biomarker of TB.

Arbuscular Mycorrhizal Fungi Mediate the Acclimation of Rice to Submergence.

Flooding is a critical factor that limits the establishment of a symbiosis between rice and arbuscular mycorrhizal fungi (AMF) in wetland ecosystems. The distribution of carbon resources in roots and the acclimation strategies of rice to flooding stress in the presence of AMF are poorly understood. We conducted a root box experiment, employing nylon sheets or nylon meshes to create separate fungal chambers that either prevented or allowed the roots and any molecules to pass through. We found that the mycorrhizal colonization rate and the expression of genes OsD14L and OsCERK1, which are involved in fungal perception during symbiosis, both increased in mycorrhizal rice roots following intermittent flooding compared to continuous flooding. Furthermore, AMF inoculation affected root morphological traits, facilitating both shallower and deeper soil exploration. Increased submergence intensity led to carbohydrate deprivation in roots, while high mycorrhizal colonization increased soil oxygen consumption and decreased the neutral lipid concentration in roots. However, mycorrhizal inoculation increased the rice photosynthesis rate and facilitated acclimation to submergence by mediating the expression of the genes OsCIPK15 and OsSUB1A to enhance rice shoot elongation and the sugar concentration in roots as a result of reduced competition for carbon between rice and AMF under different flooding conditions.

Low-Density Granulocytes Affect T-SPOT.TB Assay by Inhibiting the Production of Interferon-γ in T Cells via PD-L1/PD-1 Pathway.

BACKGROUND: T-SPOT TB (T-SPOT) assay is widely used for detection of Mycobacterium tuberculosis infection that is based on the detection of M. tuberculosis-specific interferon-γ-secreting T cells (ISCs) in peripheral blood mononuclear cells (PBMCs). Recently, high frequencies of low-density granulocytes (LDGs) were found in the PBMCs of tuberculosis patients. Whether these LDGs affect the detection of T-SPOT has not been investigated. The impact of LDGs on T-SPOT assay and related mechanism were investigated in this study. METHODS: The correlations between the frequencies of LDGs and the results of T-SPOT were analyzed. T-SPOT with LDG-removed PBMCs and PBMCs with exogenous addition of LDGs were performed. The possible mechanism was explored by detecting the levels of negative immune regulatory molecules on LDGs. The impact of programmed death ligand 1 (PD-L1) on T-SPOT was evaluated and confirmed by function blocking with neutralizing antibody. RESULTS: The positive rates of T-SPOT and ISCs in tuberculosis patients with low LDGs frequency (n = 22) were significantly higher than those with high LDGs frequency (n = 39). Removal or exogenous addition of LDGs significantly increased or decreased the ISCs and the positive rate of T-SPOT. The frequencies of interferon-γ-producing T cells were negatively correlated with the frequencies of LDGs. The expression of PD-L1 was significantly elevated on LDGs. Pretreatment of LDGs with anti-PD-L1 antibody significantly counteracted the impact of LDGs on T-SPOT. Treatment of PBMCs with anti-PD-L1 antibody resulted in comparable ISCs with that of LDG removal. CONCLUSION: LDGs can inhibit the production of interferon-γ in T cells and decrease the positive rated of T-SPOT assay via highly expressed PD-L1.

Recurrence Is a Noticeable Cause of Rifampicin-Resistant Mycobacterium tuberculosis in the Elderly Population in Jiangxi, China.

Setting: Rifampicin-resistant tuberculosis (RR-TB) in elderly people in Jiangxi Province, China. Objective: To investigate the incidence of RR-TB and risk factors in elderly people within a hospital setting in China. Design: Retrospective cohort study. Methods: A comparative study was performed to analyze RR-TB and rifampicin-susceptible TB (RS-TB). The 15-locus mycobacterial interspersed repetitive unit-variable number of tandem repeats (MIRU-VNTR) method was used to distinguish between relapse and reinfection. Results: Twenty-three recurrent cases occurred in 151 elderly patients with RR-TB, and 24 recurrent cases occurred in 466 elderly patients with RS-TB during this period. TB recurrence was significantly different in the RR-TB and RS-TB groups (OR = 0.35, 95% CI: 0.14-0.88; χ2 = 5.28, P = 0.03). Comparing the risk factors for RR-TB and RS-TB, we found that educational level, age, and pulmonary cavity were inextricably linked to RR-TB in elderly patients. Furthermore, pulmonary cavity, HIV status, and alcohol consumption were associated with recurrence in elderly RR-TB patients. Conclusions: Recurrence is an important source of RR-TB in the elderly population. It is necessary to promptly treat tuberculosis patients, prevent the spread of AIDS, and reduce alcohol intake to control recurrent RR-TB in the elderly population.

Prevalence and molecular characterization of multidrug-resistant M. tuberculosis in Jiangxi province, China.

Multidrug-resistant Mycobacterium tuberculosis (MDR-TB) is a severe health threat to human beings; however, the epidemic and molecular characteristics exist along with the change in the geographic environment and genealogy. Jiangxi province is located in southeast China, which is a high-MDR-TB burden area. Rifampin (RIF) and isoniazid (INH) are the most important first-line anti-tuberculosis drugs. The major drug target genes include rpoB for RIF and katG, inhA, and ahpC for INH. To determine the frequency and distribution of mycobacterial mutations in these genes, we sequenced specific genes of M. tuberculosis that are associated with resistance to RIF and INH in 157 phenotypic MDR isolates. At the same time, RD105 DTM-PCR and 15 loci MIRU-VNTR were performed to demonstrate the genetic lineage. It was shown that the Beijing genotype was predominant (84.1%) among these strains. The results also showed mutations within the 81 bp core region of rpoB in 93.6% of strains and mutations in a structural gene (katG) and two regulatory regions (the promoter of inhA and intergenic region of oxyR-ahpC) were shown in 88.5% of phenotypic MDR isolates. There were no significant differences in codon mutations between the Beijing and non-Beijing genotypes, as well as the clustered and no-clustered strains. The most prevalent mutations involved in RIF and INH were Ser531Leu in rpoB (55.4%) and Ser315Thr in KatG (56.1%), respectively. There was no significant difference in RIF and INH resistance between MDR-TB and other drug-resistant tuberculosis (DR-TB). The results demonstrated that some MDR-TB patients are predicted to have recent transmission.

Mycobacterium tuberculosis Infection Induces Low-Density Granulocyte Generation by Promoting Neutrophil Extracellular Trap Formation via ROS Pathway.

The roles and characteristics of low-density granulocytes (LDGs) have recently attracted attention; however, the mechanism of the formation of LDGs is yet unclear. In one of our previous studies, the frequency of LDGs was significantly elevated in the peripheral blood of tuberculosis patients, and in situ activation contributed to the generation of LDGs upon Mycobacterium tuberculosis infection. However, the underlying molecular mechanisms are yet to be elucidated. In the present study, the release of neutrophil extracellular traps (NETs) and the levels of ROS were regulated before the normal-density granulocytes (NDGs) to be infected with M. tuberculosis, and the conversion of NDGs to LDGs was monitored subsequently as well. The results showed that tuberculosis-related LDGs spontaneously released high levels of NETs. Promoting the release of NETs led to increase in the conversion of NDGs to LDGs in M. tuberculosis infection, while inhibiting the release of NETs suppressed this conversion after the infection. The M. tuberculosis infection significantly increased the ROS levels in neutrophils and the conversion of NDGs to LDGs. Scavenging ROS or blocking the ROS generation of M. tuberculosis-infected NDGs significantly suppressed the release of NETs and blocked the generation of LDGs. Moreover, inhibiting the formation of NETs without affecting the levels of ROS significantly decreased the conversion of NDGs to LDGs after M. tuberculosis infection. Overall, this study demonstrated that M. tuberculosis could induce the generation of LDGs by promoting the release of NET via ROS pathway.

Circulating circular RNAs hsa_circ_0001204 and hsa_circ_0001747 act as diagnostic biomarkers for active tuberculosis detection.

In recent years, increasing evidence has suggested that circRNAs can serve as novel diagnostic markers for many diseases. However, little is known about the value of circRNAs in the diagnosis of active tuberculosis (TB). In this study, 10 circRNAs which we previously found to be involved in Mycobacterium tuberculosis infection were selected as candidate targets for subsequent circulating circRNA assay. Compared with healthy controls, plasma levels of hsa_circ_0001204 and hsa_circ_0001747 were significantly decreased (P < 0.001). Plasma levels of hsa_circ_0001204 and hsa_circ_0001747 were correlated with TB severity. Hsa_circ_0001204 and hsa_circ_0001747 were selected for further analysis in another 145 TB patients and 120 control individuals. The area under the receiver operating characteristic curve (AUC) for distinguishing TB patients was 0.928 (95% confidence interval: 0.897-0.960; sensitivity = 86.21%, specificity = 89.17%) when hsa_circ_0001204 and hsa_circ_0001747 were used in combination. Further evaluation on potential biomarkers showed that hsa_circ_0001204 and hsa_circ_0001747 may specifically identify patients with TB. Additionally, hsa_circ_0001204 and hsa_circ_0001747 plasma levels after treatment were significantly higher than that pre-treatment (P < 0.001). Our present study indicates that circulating hsa_circ_0001204 and hsa_circ_0001747 may represent novel plasma biomarkers for TB diagnosis.

Plasma Circular RNAs hsa_circ_0001953 and hsa_circ_0009024 as Diagnostic Biomarkers for Active Tuberculosis.

Recent studies have demonstrated that circular RNAs (circRNAs) could serve as potential molecular markers for disease diagnosis; however, little is known about their diagnostic value in active tuberculosis (TB). This study first performed a microarray screening of circRNA changes in plasma samples from 3 patients with active pulmonary TB and 3 healthy controls. Then, candidate circRNAs were selected for validation on a quantitative real-time PCR system. Of the 61 differentially expressed circRNAs recorded, 43 and 18 were upregulated and downregulated in the TB group, respectively. Validation assays demonstrated that plasma levels of 6 circRNAs, including hsa_circ_0009024, hsa_circ_0001953, hsa_circ_0008297, hsa_circ_0003528, hsa_circ_0003524 and hsa_circ_0015879 were remarkably increased in TB patients. Plasma levels of hsa_circ_0001953 and hsa_circ_0009024 were correlated with TB severity. Next, hsa_circ_0001953 and hsa_circ_0009024 were assessed in an independent cohort consisting of 120 TB patients and 100 control individuals. An area under the receiver operating characteristic (ROC) curve of 0.915 (95% confidence interval 0.880-0.951; P < 0.001) was obtained for detecting TB, with hsa_circ_0001953 and hsa_circ_0009024 used in combination. Additionally, plasma levels of hsa_circ_0001953 and hsa_circ_0009024 were reduced significantly in patients after treatment (P < 0.001). The present findings indicate that the circRNAs hsa_circ_0001953 and hsa_circ_0009024 may represent novel plasma biomarkers for active TB diagnosis.

Variable-number tandem repeat markers for Mycobacterium intracellulare genotyping: comparison to the 16S rRNA gene sequencing.

INTRODUCTION: Characterizing Mycobacterium intracellulare responsible for nontuberculous mycobacterial (NTM) infections may aid in controlling outbreaks. This study aimed to compare 16S ribosomal ribonucleic acid (rRNA) sequencing and variable-number tandem repeat (VNTR) genotyping of M. intracellulare strains isolated from clinical samples, and to characterize VNTR clusters associated with NTM infections or cavity formation. METHODOLOGY: Sputum samples were obtained from 77 HIV-negative patients with pulmonary disease between 2009 and 2013. One M. intracellulare strain was isolated from each patient and genotyped using 16S rRNA and eight loci VNTR sequencing. RESULTS: Single nucleotide polymorphism (SNP) genotyping identified seven point mutations at nucleotide positions 101, 178, 190, 252, 382, 443, and 490 in 16S rRNA, and four SNP patterns were identified: type 1 (16 strains), 2 (41 strains), 3 (11 strains), and 4 (1 strain); 5 strains had unique SNP patterns. VNTR genotyping identified VNTR12 as the most discriminating marker (allelic diversity 0.692). VNTR3 was the most homogeneous marker (allelic diversity 0.518), but each locus had high discriminating ability. The 77 strains were clustered according to the unpaired group method using arithmetic averages: cluster 1 (17 strains), 2 (43 strains), 3 (9 strains), and 4 (4 strains); 4strains had unique SNP patterns. Overall, over 90% strains were matched to similar SNP and VNTR groupings. VNTR clusters were associated with NTM infection (p =0.007) and presence of a cavity (p =0.042). Both methods distinguished four subtypes of M. intracellulare, which corresponded. CONCLUSIONS: VNTRs may represent an effective, user-friendly, low-cost typing technique.

Identification of differentially expressed circular RNAs in human monocyte derived macrophages response to Mycobacterium tuberculosis infection.

Macrophages act as the first line of host immune defense against Mycobacterium tuberculosis (Mtb). Recent studies have demonstrated circular RNAs (circRNAs) are implicated in a variety of physiological and pathological processes; however, the role of circRNAs in macrophages response to Mtb infection remain unknown. To address this issue, here we characterized circRNAs expression profiles in human monocyte derived macrophages (MDMs) response to Mtb infection using microarray assay. Our results revealed that many circRNAs were differentially expressed in human MDMs after Mtb infection; of these, 32 circRNAs were up-regulated and 110 were down-regulated. Real time PCR results were generally consistent with the microarray data. Furthermore, we found that hsa_circ_0043497 and hsa_circ_0001204 may be effective diagnostic biomarkers for TB. This study provides the first evidence that circRNAs alterations are involved in human MDMs response to TB infection and reveal potential targets for diagnostics and the treatment of TB.

EBP50 induces apoptosis in macrophages by upregulating nitric oxide production to eliminate intracellular Mycobacterium tuberculosis.

Mycobacterium bovis BCG is known to have the capacity to inhibit the positioning of iNOS on BCG-containing phagosomes by interfering with EBP50, a scaffolding protein that controls the recruitment of inducible nitric oxide synthase (iNOS) at the vicinity of phagosomes in macrophages. However, knockdown of the expression of EBP50 still facilitates the intracellular survival of BCG, which suggested that EBP50 may have some other unknown antimycobacterial properties. In this study we show that overexpression of EBP50 by a recombinant lentivirus had no effect on the iNOS recruitment to M.tuberculosis-containing phagosomes, but significantly promoted the elimination of intracellular M.tuberculosis. We revealed in the present study that the enhancement of intracellular killing to M. tuberculosis upon EBP50 overexpression was due to the increased level of apoptosis in macrophages. We showed that EBP50 overexpression significantly increased the expression of iNOS and generation of nitric oxide (NO), and EBP50-induced apoptosis was NO-dependent and mediated by Bax and caspase-3. We found that M. tuberculosis decreases while Mycobacterium smegmatis increases the expression of EBP50 in RAW264.7 cells, which suggested that virulent mycobacteria are capable of modulating the antimycobacterial properties of macrophages by inhibiting the expression and interfering with the function of EBP50.

Low-Density Granulocytes Are Elevated in Mycobacterial Infection and Associated with the Severity of Tuberculosis.

Tuberculosis remains a global health problem caused by infection with Mycobacterium tuberculosis. Numerous studies have established a close correlation between the development of tuberculosis and the roles of neutrophils. Recently, a distinct population of CD15+ granulocytes was found to be present in the peripheral blood mononuclear cell (PBMC) fraction in humans. This population of granulocytes, termed low-density granulocytes (LDGs), was reported to be elevated and associated with disease activity or severity in a number of different conditions including SLE, asthma and HIV infection. However, both the frequency and clinical significance of LDGs associated with tuberculosis are unclear. Here we determined LDG levels and made comparisons between subjects with active pulmonary tuberculosis (PTB) and healthy controls, between PTB patients with mild-to-moderate disease and patients with advanced disease, and among PTB patients following anti-tuberculous therapy of varying durations. The direct correlation between M. tuberculosis infection and LDG levels was confirmed by in vitro infection of whole peripheral blood and isolated granulocytes with mycobacteria. Our results demonstrated that PBMCs in PTB patients contained significantly elevated percentages of LDGs compared with control subjects. LDGs in tuberculosis expressed higher levels of activation markers compared to normal-density granulocytes (NDGs). M. tuberculosis induced the generation of LDGs in both whole blood and isolated NDGs from control subjects, which suggests that LDGs associated with M. tuberculosis infection are likely to originate from in situ activation. Furthermore, our results revealed that the frequency of LDGs is associated with the severity of tuberculosis.

Mycobacterium tuberculosis-Induced Polarization of Human Macrophage Orchestrates the Formation and Development of Tuberculous Granulomas In Vitro.

The tuberculous granuloma is an elaborately organized structure and one of the main histological hallmarks of tuberculosis. Macrophages, which are important immunologic effector and antigen-presenting cells, are the main cell type found in the tuberculous granuloma and have high plasticity. Macrophage polarization during bacterial infection has been elucidated in numerous recent studies; however, macrophage polarization during tuberculous granuloma formation and development has rarely been reported. It remains to be clarified whether differences in the activation status of macrophages affect granuloma formation. In this study, the variation in macrophage polarization during the formation and development of tuberculous granulomas was investigated in both sections of lung tissues from tuberculosis patients and an in vitro tuberculous granuloma model. The roles of macrophage polarization in this process were also investigated. Mycobacterium tuberculosis (M. tuberculosis) infection was found to induce monocyte-derived macrophage polarization. In the in vitro tuberculous granuloma model, macrophage transformation from M1 to M2 was observed over time following M. tuberculosis infection. M2 macrophages were found to predominate in both necrotic and non-necrotic granulomas from tuberculosis patients, while both M1 and M2 polarized macrophages were found in the non-granulomatous lung tissues. Furthermore, it was found that M1 macrophages promote granuloma formation and macrophage bactericidal activity in vitro, while M2 macrophages inhibit these effects. The findings of this study provide insights into the mechanism by which M. tuberculosis circumvents the host immune system as well as a theoretical foundation for the development of novel tuberculosis therapies based on reprogramming macrophage polarization.

Spatial resolution and information transfer in scanning transmission electron microscopy.

The relation between image resolution and information transfer is explored. It is shown that the existence of higher frequency transfer in the image is just a necessary but not sufficient condition for the achievement of higher resolution. Adopting a two-point resolution criterion, we suggest that a 10% contrast level between two features in an image should be used as a practical definition of resolution. In the context of scanning transmission electron microscopy, it is shown that the channeling effect does not have a direct connection with image resolution because sharp channeling peaks do not move with the scanning probe. Through a quantitative comparison between experimental image and simulation, a Fourier-space approach is proposed to estimate defocus and sample thickness. The effective atom size in Z-contrast imaging depends on the annular detector's inner angle. Therefore, an optimum angle exists for the highest resolution as a trade-off between reduced atom size and reduced signal with limited information transfer due to noise.

HAADF-STEM imaging with sub-angstrom probes: a full Bloch wave analysis.

A full coherent Bloch wave calculation is presented to investigate high-angle annular dark-field image formation for sub-angstrom probes in scanning transmission electron microscopy (STEM). With increasing illumination angle, the contribution of the 1s bound state increases to a maximum at an optimum probe angle, after which we find increasing contributions from high-angle plane wave states around the periphery of the objective aperture. Examination of image contributions from different depths within a crystal shows an oscillatory behavior due to the beating between 1s and non-1s states. The oscillation period reduces with decreasing probe size, while the relative contribution from a specific depth increases. This signifies a changeover from a projection mode of imaging to a depth-slicing mode of imaging. This new mode appears capable of resolving three-dimensional atomic structures in future generation aberration-corrected STEM.