Human cytomegalovirus (HCMV) usually establishes a lifelong latent infection after primary infection. Reactivation occurs sporadically and is controlled by cell-mediated immune response (CMIR). Monitoring of CMIR against CMV is mandatory in immunocompromised patients to adjust immunosuppressive drugs to prevent serious end-organ damage after CMV reactivation. Intracellular staining (ICS) and enzyme-linked immunospot (ELISPOT) quantifying CMV-specific T-cells are generally used as surrogate markers for CMIR against CMV. Whether the results of these 2 methods correlate well is not known. This study compared the numbers of CMV-specific T-cells identified by ELISPOT and ICS in healthy adult volunteers. Correlation with CMV serological status was explored. Thirty peripheral blood samples from healthy individuals were quantified for IFN-γ-producing cells after stimulation with whole CMV and IE1 using ICS, and IFN-γ-secreting cells after stimulation with whole CMV lysate and IE1 peptide pool using ICS and IFN-γ-secreting cells by ELISPOT. Anti-CMV IgG levels were analyzed concomitantly using a chemiluminescent microparticle immunoassay. There were 30 healthy participants, 15 (50%) male, with a mean age of 37.8 (± 7.6) years. Twenty-eight (93.3%) were seropositive against CMV. The CMV-specific CD3+ cells, as measured by ICS, were highly correlated with the spot numbers obtained by the ELISPOT, irrespective of CMV antigens used (whole CMV, r = 0.7677, p < 0.0001; IE1, r = 0.6516, p < 0.0001). The numbers of CMV-specific CD3+ cells quantified by IE1 stimulation by these 2 assays were statistically correlated with anti-CMV IgG levels (ICS, r = 0.5070, p = 0.004; ELISPOT, r = 0.4384, p = 0.015). Among the 30 participants, CMV-specific T cells were detected in all participants (100%), including the two seronegative individuals. The present study demonstrated that CMV-specific T-cells measured by ICS and ELISPOT assays were well correlated, suggesting that these assays could be used to monitor CMV-specific T-cells. CMV IgG levels may reflect prior CMV infection and CMV-specific cell-mediated immunity (CMIR) in immunocompetent individuals.
Resistance spot welding (RSW), which is still extensively used in the automobile, aerospace, and other equipment production industries, depends critically on the quality of the weld. The size of the welded joint nucleus diameter was used as the output of the spot welding quality monitoring model, with welding current, welding voltage, welding air pressure, and welding time selected as input parameters. This study developed a neural network-based online monitoring model for resistance spot welding. The Harris Hawk Herd Algorithm Optimized BP Neural Network (HHOBP) neural network model, which is the basis for the spot welding quality prediction model, has an R2 coefficient value of up to 0.9981 and may somewhat predict the quality of welded connections. Compared with PSO-BP, GWO-BP, CS-BP, GA-BP, traditional BPNN and support vector machine models, HHOBP has achieved 93.06% ~ 96.15% (MAE), 99.43% ~ 99.81% (MSE), 92.18% ~ 95.46% (RMSE) and 87.54% ~ 96.83% (MAPE) in different error indicators, reflecting excellent modeling capabilities.
The World Health Organization recommends screening and providing preventive treatment of tuberculosis in high-risk groups. We compared the performance of the latest versions of two widely used interferon (IFN)-γ release assays (IGRAs), the QuantiFERON-TB Gold Plus chemiluminescent assay (QFT-Plus CLIA) and the semi-automated T-SPOT.TB assay, and assessed the potential benefit of a dual-testing strategy in immunosuppressed patients and/or candidates for biological therapy. From September 2022 to December 2023, we included 3748 immunosuppressed patients. IGRAs were performed according to manufacturers' instructions. Agreement was evaluated using Cohen's kappa (κ), and subgroup analyses were stratified by age and underlying disease. At least one IGRA was positive in 13.4% of patients (12.0% QFT-Plus CLIA; 9.9% T-SPOT.TB). Indeterminate results were infrequent (1.8% QFT-Plus CLIA; 2.6% T-SPOT.TB). Overall agreement was high (93.0%; κ=0.67), but lower among IGRA-positive cases (63.9%; κ=0.62). Discrepancies occurred in 7.0% of patients, most frequently in children (0-14 years: 15.4%), older adults (≥75 years: 13.8%), individuals living with HIV (38%), rheumatoid arthritis/Sjögren's syndrome (12.4%) and other chronic inflammatory diseases, transplant recipients or immunodeficiencies (11.9%). Among IGRA-positive cases (n=502), most discordant results corresponded to QFT-Plus CLIA-positive/T-SPOT.TB-negative patterns (131 of 181; 72.4%). In immunosuppressed patients, dual IGRA testing enhances diagnostic sensitivity and can strengthen clinical decision-making. Concordant results increase diagnostic certainty, whereas discordant findings gain interpretative value when contextualised by disease group and assay-specific behaviour IGRA.
This study evaluates the drivers of radionuclide spatial heterogeneity in topsoil of the Lake Sevan Basin (Armenia) - the largest high-mountain freshwater body in the South Caucasus - using integrated statistical and spatial analyses. A total of 170 soil samples were analyzed for Ra-226, Th-232, K-40, Cs-137, and gross beta activity, together with in-situ dose was measurement. Statistical analysis revealed strong inter-correlations among natural radionuclides (with Spearman's test) and bimodal distribution patterns for Th-232 and gross beta activity, indicating the presence of distinct source domains. Land-use analysis confirmed significant effects on K-40 and Cs-137 distributions, with higher Cs-137 in low-disturbance soils and lowest in arable land, whereas K-40 was notably elevated in forest soils. The spatial heterogeneity of these parameters was modeled using geostatistical methods including Empirical Bayesian Kriging (EBK) and Getis-Ord Gi* (Gi*) Hot Spot analysis. The analysis identified statistically significant hot spots of Ra-226, Th-232 and radium equivalent activity in southwestern, volcanic-rock-dominated part of the basin, and cold spots in lacustrine-derived soils in the eastern area. K-40 displayed additional enrichment in northwestern agricultural zones, suggesting a potential contribution from long-term agricultural practices. Cs-137 exhibited limited but distinct hot-spot clustering in minimally disturbed soils, reflecting post-depositional fallout processes. Radiological risk assessment revealed Ra-226 and Th-232 as the main risk contributors for the southwestern located settlements. The study demonstrates that integrating statistical inference, spatial modelling, and radionuclide geochemistry enables process-based interpretation of soil radioactivity and provides a transferrable framework for contamination assessment, radioecological targeted monitoring, and management prioritization in complex mountain environments.
Climate change poses significant challenges to aquaculture through rising temperatures and salinity fluctuations, necessitating insights into molecular adaptation mechanisms in resilient species like pearl spot (Etroplus suratensis) which is an euryhaline fish. This study performed de novo transcriptomic profiling of gill, liver, and kidney tissues from juvenile pearl spot exposed to combined temperature (28 °C control, 31 °C intermediate, 34 °C stress) and salinity (0 ppt freshwater, 15 ppt brackishwater) treatments over 90 days in triplicate setups. Illumina NovaSeq 6000 sequencing generated 1634 million raw reads, yielding 1613 million high-quality paired-end reads post-filtering. De novo assembly produced 876,873 contigs enabling identification of 23,204 differentially expressed genes in liver, 11,190 in gill, and 8005 in kidney, revealing tissue-specific responses dominated by temperature with synergistic salinity effects. Liver exhibited robust metabolic remodelling (upregulated FASN, ACC, HK4 for lipid/glucose pathways), proteostasis (HSP70/90, HYOU1), balanced immunity (C3 downregulation, dcst1/hells upregulation), and transcription/apoptosis regulation (MafA-like, RNF2). Gill DEGs emphasized osmoregulation (Na+/K+-ATPase downregulation, Piezo2 upregulation), oxygen transport (hemoglobin subunits), and ECM remodelling. Kidney highlighted ion homeostasis (STC, CEBPD) and DNA repair. Gene Set Enrichment Analysis revealed enriched pathways including fatty acid biosynthesis (27-fold at T3L), steroid biosynthesis, arginine metabolism, and oxidative phosphorylation. Quantitative RT-PCR validation of six key DEGs confirmed RNA-seq results (R² = 0.9). These findings elucidate tissue-specific molecular signatures characterising pearl spot resilience, identifying biomarkers (HSPs, STC) for breeding climate-resilience aquaculture strains in response to environmental shifts.
During the 2020 through 2024 growing seasons, pepper fruits exhibiting symptoms typically associated with spotted wilt were observed in the Arica y Parinacota region, in northern Chile. As this region is a key winter supplier of vegetables to southern Chile, the present study aimed to identify and characterize orthotospoviruses infecting local pepper crops. Species-specific and generic primers targeting orthotospoviruses were used in RT-PCR assays for virus identification, and thrips vectors were characterized both morphologically and by PCR using ITS- and MtCOI-specific primers. Tomato spotted wilt virus (TSWV; Orthotospovirus tomatomaculae) and the putative new orthotospovirus, pepper necrotic spot virus (PNSV) were detected in samples collected from four production fields in Arica y Parinacota. Among the known orthotospovirus vectors, only Frankliniella occidentalis was identified at sampled sites. While TSWV is already endemic in Chile, this represents the first report of PNSV in the country. Given that PNSV has been previously reported in Peru, the geographic proximity of Arica y Parinacota raises concern for its continued southward spread into Chile. This finding underscores the need for further epidemiological studies on PNSV and the development of strategies to mitigate its dissemination within the country.
Congestion is the dominant driver of hospitalization in acute heart failure and relief of congestion remains a central therapeutic target. Loop diuretics are first-line therapy for decongestion, yet their dosing and escalation in routine practice often rely on subjective bedside assessment, variable urine output thresholds, weight change, and delayed laboratory trends approaches that incompletely capture the primary pharmacodynamic goal of diuretic therapy: natriuresis. Neurohormonal activation and renal sodium avidity in heart failure can lead to poor diuretic response despite apparently adequate diuretic dosing, and persistent congestion is consistently associated with adverse outcomes. In this context, early measurement of spot urinary electrolytes especially urinary sodium, and potentially urinary chloride and urinary creatinine offers an objective, rapid method to quantify natriuretic response and identify inadequate decongestion early enough to adjust therapy. This review summarizes the pathophysiologic basis linking renal sodium handling to congestion, critiques traditional metrics used to titrate diuretics, synthesizes the clinical evidence supporting spot urinary sodium-based assessment (observational cohorts and emerging randomized/protocolized strategies), and outlines pragmatic implementation considerations, including confounders such as chronic kidney disease, concomitant SGLT2 inhibitors, and timing of sampling. While natriuresis-guided strategies reliably improve natriuresis and process-of-care metrics, definitive evidence for improved hard outcomes remains evolving, underscoring the need for standardized protocols and larger outcomes trials.
During orthodontic treatment, fixed orthodontic appliances complicate optimal oral hygiene maintenance, frequently causing white spot lesions (WSLs). Clinically, WSLs are typical chalky-white opacities on the enamel surface, recognized as the early reversible stage of dental caries. In this study, a novel antibacterial and remineralizing material-CEN composites (Chitosan-Ca3(PO4)2, CS-CaP) microgels loaded with EGCG nanoparticles (NPs)-was designed to treat enamel white spot lesions. Material characterization experiments were conducted on EGCG-NPs, chitosan-calcium phosphate microgels and CEN composites; the dialysis bag diffusion method was used to determine the composite drug release rate. Enamel blocks were prepared, an acid-etched demineralization model was established, and in vitro remineralization experiments were performed to screen the optimal CEN composite concentration for enamel remineralization and confirm the synergistic effect of EGCG-NPs on remineralization. The CCK-8 method was employed to assess the cytotoxicity of the CEN complex. Material characterization experiments confirmed the successful synthesis of CEN composites. Drug release tests showed EGCG-NPs release rate reached 50% at 10 h and 80% at 70 h. In vitro remineralization experiments confirmed that the CEN-5 group was the optimal concentration for remineralization. In vitro experiments of the CEN-5 group demonstrated that EGCG-NPs significantly enhanced the gel's remineralization efficacy. The CCK-8 assay demonstrated that the CEN composite possessed no cytotoxic effects.
White spot lesions (WSLs) are a common adverse outcome of fixed orthodontic treatment, prompting the search for salivary microbial predictors for early risk identification. Sixty-four patients undergoing fixed metal self-ligating orthodontic treatment were categorized into WSL and non-WSL groups based on lesion development after 12 months. Clinical indices (DI-S, CI-S, OHI-S, GI) and unstimulated whole saliva samples were collected at baseline and after 12 months. The salivary microbiome was analyzed using 16S rRNA gene sequencing. The prevalence of WSLs was 54.7% at the patient level. Clinical indices did not differ significantly between groups. Although overall microbial community structure was similar at baseline, several taxa were enriched in patients who later developed WSLs, including Actinomyces, Rothia and Granulicatella. After 12 months, the WSL group showed reduced phylogenetic diversity and a microbial profile enriched in acidogenic and anaerobic taxa. Functional prediction indicated increased carbohydrate metabolism and fermentative pathways in WSL patients. WSL development during fixed orthodontic treatment was associated with baseline salivary microbial differences and treatment-related ecological shifts. Salivary microbiome profiling may help identify patients at higher risk and support early risk stratification and preventive strategies in orthodontic care.
This in vitro study aimed to evaluate the accuracy and bias of Ultrasound (US) in measuring the depth of white spot lesions (WSLs), in comparison to micro-CT (µ-CT). The study included 120 bovine maxillary incisors. Artificial WSLs were created on the facial surface of each incisor. Incisors were categorized into two groups, shallow WSLs and deep WSLs and subjected to 2-day and 4-day pH cycling protocols, respectively. All samples were imaged with both µ-CT and US using second harmonic imaging at 12/24 MHz. WSL depth was measured and the average calculated. Enamel thickness was assessed on the µ-CT images using the same methodology. Mean lesion depths in the shallow WSLs group were 138.3±17.8 μm (µ-CT) and 169.2 ± 37.8 μm (US), while in the deep WSLs group, depths averaged 299.9 ± 47.7 μm (µ-CT) and 309.3 ± 75.7 μm (US). The absolute mean differences between µ-CT and US were significantly different from zero (p < 0.001). US consistently overestimated lesion depth compared to µ-CT in both groups, with a significant difference in the shallow WSLs group (p < 0.001). US demonstrates lower accuracy than µ-CT in measuring WSL depth, consistently overestimating lesion depths, particularly in the shallow WSLs group. The ability of a diagnostic method to measure WSLs of varying depths is clinically important, as treatment efficacy depends on lesion depth and the extent of enamel demineralization. US in non-invasive and demonstrates potential for clinical use; however, further validation is required before it can be reliably applied to the clinical assessment of WSL depth. Clinicians must also consider practical factors such as equipment cost and size, the need for coupling materials, and requirements for operator training and calibration.
To present three cases where the retrobulbar spot sign (RBSS) was observed, but central retinal artery occlusion (CRAO) was not confirmed. Transorbital sonography was employed to detect the presence of RBSS in all patients. RBSS was detected in all three patients. However, only one patient had CRAO, with RBSS-positive findings in both eyes, although one eye showed normal function. The other two patients exhibited RBSS without evidence of CRAO. RBSS remained unchanged in all four eyes during the follow-up period. RBSS may not be specific to CRAO, as it was observed in non-CRAO cases. Further research is necessary to clarify its diagnostic significance and differentiate it from other pathologies.
Bisphenol analogues, widely used in consumer products, are emerging environmental contaminants with documented endocrine-disrupting effects. Conventional biomonitoring methods relying on venous blood or urine are logistically challenging, especially for vulnerable populations. Dried blood spot (DBS) sampling serves as a minimally invasive and logistically superior alternative, yet its application for trace-level bisphenol analogues analysis remains limited due to sensitivity issues, background contamination, and lack of standardization. Herein, we developed a high-throughput, kit-based method for the simultaneous quantification of 16 bisphenol analogues in DBS samples using protein precipitation combined with phospholipid removal, followed by UHPLC-MS/MS analysis. The method demonstrated high sensitivity, with method detection limits (MDLs) ranging from 0.04 to 1.17 μg/L, method quantitation limits (MQLs) ranging from 0.16 to 1.48 μg/L, and excellent accuracy and precision (recoveries: 70-128%; RSDs < 20%). Matrix effects were effectively compensated using stable isotope-labeled internal standards. The method's application to 33 authentic DBS samples indicated substantial exposure to bisphenol F (BPF), bisphenol A (BPA), bisphenol S (BPS), and diphenyl sulfone (DPS). This standardized, scalable workflow provides a robust tool for large-scale biomonitoring and epidemiological studies of bisphenol analogues.
Therapeutic drug monitoring (TDM) of psychotropic agents is essential for optimizing therapeutic efficacy and minimizing adverse effects. However, conventional liquid chromatography-tandem mass spectrometry (LC-MS/MS) workflows are poorly suited for point-of-care applications due to labor-intensive sample preparation, prolonged turnaround times, and dependence on centralized analytical laboratories. Here, we present a rapid, on-site analytical platform that integrates dried blood spot (DBS) sampling with a modified capillary-in-capillary electrospray ionization (CC-ESI) source coupled to a miniature mass spectrometer (mini-MS). The sampling capillary was re-engineered as a disposable polypropylene microtube capable of accommodating a DBS punch and extraction solvent, thereby enabling fully integrated online extraction, ionization, and mass analysis within minutes. Critical parameters, including infrared drying duration and extraction solvent composition, were systematically optimized. The method demonstrated excellent linearity (R2 > 0.95) and precision for six widely prescribed psychotropic drugs: diazepam, flurazepam, clozapine, zolpidem, risperidone, and methaqualone. Analytical performance was comparable to that achieved using conventional solid-phase extraction, yet sample preparation time was reduced from several hours to mere minutes. Clinical utility was preliminarily demonstrated by the successful detection and confirmation of clozapine and aripiprazole in blood samples from patients receiving long-term pharmacotherapy. This DBS-CC-ESI-mini-MS platform represents a promising advance toward decentralized TDM of psychotropic drugs, offering minimal invasiveness, operational simplicity, and rapid, reliable quantitative capability.
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[This corrects the article DOI: 10.1107/S1600576724004333.].
Cryptococcal antigen (CrAg) lateral flow assays (LFA) are widely used for the rapid diagnosis of cryptococcal meningitis because of their excellent sensitivity and specificity. We report a rare case of culture- and sequencing-confirmed Cryptococcus gattii VGI meningitis in an immunocompetent 43-year-old man with persistently negative IMMY CrAg LFA and semi-quantitative CrAg SQ results in both cerebrospinal fluid (CSF) and serum. The patient presented with fever, chronic progressive headache, diplopia, and papilledema. CSF analysis demonstrated lymphocytic pleocytosis, hypoglycorrhachia, elevated protein, and budding yeast cells on direct microscopy and India ink preparation. BioFire FilmArray meningitis/encephalitis panel detected C. neoformans/gattii species complex, while fungal culture and sequencing confirmed C. gattii VGI. False-negative CrAg LFA results persisted despite serial sample dilutions up to 1:2560. This case highlights a rare diagnostic pitfall of CrAg testing and underscores the importance of integrating microscopy, culture, and molecular methods when clinical suspicion remains high despite negative antigen assays.
Proteins that bind to a target protein of interest, termed "binders", are essential components of biological research reagents and therapeutics. Target proteins present multiple binding surfaces with varying interaction potential. "Hot spots", or high-potential surfaces, are experimentally identified as the most probable binding sites in de novo discovery campaigns. However, hot spots and their default binding modes do not always confer the desired specificity. Related proteins or isoforms often share similar hot spots, resulting in promiscuous binding. Interaction with a hot spot may also fail to elicit the intended biological outcome. Consequently, methods that direct de novo binder discovery toward targets with defined specificity are critically needed. We recently developed phage-assisted noncontinuous selection of binders (PANCS-Binders), a selection platform with unparalleled speed and sequence-function fidelity that enables routine de novo binder discovery within days. However, because PANCS-Binders selections enrich variants based primarily on affinity, secondary screening is unlikely to identify binders to lesser hot spots because of the high likelihood of convergence. These alternative binding surfaces with weaker inherent interactions may possess desirable specificity profiles. Here, we develop PANCS-spec-Binders, which incorporates simultaneous selection and counterselection to control the specificity of enriched binders. We demonstrate PANCS-spec-Binders in two proof-of-concept applications: 1) discovery of isoform-selective binders that bind HRAS with >100-fold higher affinity than the highly related KRAS isoform, and 2) discovery of epitope-specific binders that either target or avoid the LIR interaction region of LC3B. PANCS-spec-Binders enables rapid identification of binders with defined specificity within days.
Food hygiene is a critical but underemphasized link in fecal-oral disease transmission in dense informal settlements. A mesh-walled meatsafe, designed to exclude insects, animals, and young children while allowing ventilation, was distributed in a randomized controlled trial in Korail, Dhaka. Despite near-universal use (87.7-99.2% of intervention households storing cooked food across five spot-check rounds), the trial detected no reductions in food contamination or child diarrhoea. This qualitative substudy investigated what technological, psychosocial, and contextual factors limited the meatsafe's protective potential despite high observed use. Guided by the Integrated Behavioral Model for Water, Sanitation, and Hygiene (IBM-WASH), we conducted 16 in-depth interviews with primary caregivers, purposively sampled to capture variation in meatsafe condition, maintenance, and food contamination profiles, and triangulated findings against spot-check records. Transcripts were thematically coded across IBM-WASH domains. Barriers preventing correct device use (fidelity barriers) included a steam-spoilage belief leading caregivers to cool food uncovered before meatsafe placement, rexine liner warping reinforcing cooling delays, and cumulative hardware friction from frame instability, latch weakness, and size constraints. Barriers that operated outside the scope of the intervention (beyond-fidelity barriers) included energy scarcity enforcing batch cooking with extended storage, monsoon flooding contaminating cooking spaces, seasonal displacement disrupting routines, and contested childcare authority limiting maternal protective behavior. The uncovered cooling period was invisible to spot-check monitoring yet represented the highest-risk contamination window. Near-universal meatsafe use without contamination reductions is explained by a behavioral gap invisible to spot-check monitoring: households stored food only after an uncovered cooling period. The meatsafe addressed only one F-diagram pathway; other fecal-oral routes remained uncontrolled. Sustainable reduction of foodborne enteric disease requires co-designed hardware addressing the full causal pathway, behavior change communication directly confronting the steam-spoilage belief, and concurrent structural investment in energy access, drainage, and pest control.
Despite improvements in early detection, Tennessee ranks among the top ten states for breast cancer mortality among women. Mammography screening and early diagnosis are critical to reducing mortality, yet access and uptake vary widely across the State. We tested whether access to diagnostic services affected screening in the State. We merged CDC PLACES estimates of screening among women aged 50-74 with geocoded FDA-certified mammography facilities to derive drive-time categories. Tract covariates included education, poverty, insurance, race/ethnicity, primary care provider density, urban/rural status, and Appalachian designation. We mapped hot/cold spots using Getis-Ord Gi* and fit four nested beta-regression models: geography only, socioeconomic only, combined, and combined plus log-provider density. Median screening prevalence was 74.8%. Hotspots clustered around Memphis, Nashville, and Knoxville; cold spots in Central Appalachian and western floodplain tracts. Geography explained ~ 23% of between-tract variation (pseudo-R2 = 0.238); socioeconomic covariates ~ 61% (0.607). In combined models, drive-time and Appalachian effects attenuated. Urban tracts screened 0.72 percentage points (pp) higher than rural, and each 1-pp rise in adults without a high-school diploma predicted - 0.13 pp. Urbanicity and education were the primary drivers of mammography screening disparities in Tennessee. To improve early diagnosis and outcomes, interventions should prioritize educational outreach, poverty reduction, insurance expansion, and improved geographic access, via mobile units or telehealth, targeting remote cold-spot tracts.
Spatial transcriptomic technologies are promising tools for elucidating fine anatomical profiles of tissues. For methods that rely on deterministic probe arrays, balancing spatial resolution, cost, and transcript-capture sensitivity is crucial to advancing spatial transcriptomics in both basic research and clinical applications. Here we present Well-ST-seq, a near-cellular-resolution platform that integrates microwell-assembled hydrogel bead carriers with combinatorial microfluidic indexing to generate predefined spatial barcode arrays. In this design, orthogonal microchannels are used for coordinate indexing, while probe construction is confined to bead carriers and executed through a simplified workflow, minimizing the need for biochemical processing inside narrow channels. Consequently, spatially barcoded capture arrays can be prepared in ∼2 h at a direct consumables cost of ∼$0.05-$0.54 per mm2 for 30-10 μm spot sizes. Using 10 μm arrays, Well-ST-seq achieves high transcript recovery in mouse hippocampus, yielding 3,896 UMIs per spot, and supports the delineation of layered organization and region-specific expression programs. Across consecutive sections of developing mouse embryonic brain, our method further enables coherent alignment and consistent domain-level mapping throughout the series. Together, Well-ST-seq provides a promising route to rapid, cost-efficient fabrication of deterministic spatial transcriptomics slides and to near-cellular spatial tissue profiling.