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Spatial Transcriptomics Inc visium spatial transcriptomics sequencing

Single‐cell and spatial transcriptome landscape of healthy and fibrotic kidneys after unilateral ischemia‐reperfusion injury (UIRI). a) Schematic representation of single‐cell RNA <t>sequencing</t> (scRNA‐seq) and spatial <t>transcriptomics</t> (ST) of kidneys from the sham and 10‐day UIRI mice, graphically designed with Biorender ( https://www.biorender.com/ ). b) t‐SNE plot illustrating the intricate cellular diversity in fibrotic kidneys, demonstrating distinct clusters representing glomerular endothelial cells (GEC), podocytes (Podo), mesangial cells (Mesa), Bowman's capsule epithelium (BC), proximal tubules (PT), descending limbs of Henle (DLOH), ascending limbs of Henle (ALOH), distal tubules (DT), principal cells (PC), intercalated cells (IC), fibroblasts (Fib), smooth muscle cells (SMC), extraglomerular endothelial cells (EGEC), monocytes (Mono), dendritic cells (DC), macrophages (Mϕ), plasmacytoid dendritic cells (pDC), proliferating mononuclear lineage (Prolif mono_L), and neutrophils (Neu), B cells (B), T cells (T), proliferating T cells (prolif T), and natural killer cells (NK). These cell types were further categorized into four major compartments: Glomerular, Renal, Interstitium, and Immune, as indicated by color grouping in the plot. c) Bubble plot illustrating the relative proportions of major kidney cell types in sham and UIRI samples. Each dot represents the proportion of a given cell type in a specific sample group, with dot size corresponding to its relative proportion. d) A comprehensive heatmap depicting the unique marker gene signature of major renal cell types. e) UMAP plot illustrating the inferred renal cell region distribution based on integrated spatial transcriptomics data from normal (Sham) and UIRI 10D mouse kidneys, generated using the 10x Genomics <t>Visium</t> platform. The identified regions include glomerular cells (Glom), distinct segments of the proximal tubule (PTS1, PTS1S2, PTS2), injured proximal tubules (InjPT), ascending limbs of Henle in cortex (ALOH(C)), distal tubules (DT), connecting tubules and collecting ducts (CNT_CD), cells at the corticomedullary junction (CMJ), fibrogenic niche regions (Niche1, Niche2), the inner stripe of the outer medulla (IOM), inner medulla (IM), renal capsule (RC), and perirenal tissue (Perirenal). f) Spatial maps illustrating the anatomical distribution of renal cell regions in Sham and UIRI 10D mouse kidneys. Region colors correspond to the classifications defined in panel (e). g) Bubble plot illustrating the relative proportions of major renal cell regions in spatial transcriptomics data from sham and UIRI 10D mouse kidneys. h) Bubble plot depicting the expression patterns of marker genes across distinct renal cell regions in spatial transcriptomics data. Dot color indicates the average gene expression level within each region, while dot size represents the proportion of spatial spots expressing the gene. i) Schematic diagram of nephron segmentation by cell types. j) Comparison of kidney anatomical regions and spatial transcriptomic clusters, showing clusters in kidney tissue (top) and the corresponding Visium H&E‐stained section (bottom). k) Renal tissue structure alterations at the corticomedullary junction (CMJ) in UIRI samples, showing the formation of two distinct fibrogenic niches, Niche1 and Niche2. l) A heatmap showing the deconvolution scores of cell type compositions across different regions in Visium spatial transcriptomics data, obtained using the RCTD method. m) Spatial FeaturePlots of RCTD‐derived cell type scores in the sham (top) and UIRI (bottom) groups, with paired panels sharing a common legend.

Visium Spatial Transcriptomics Sequencing, supplied by Spatial Transcriptomics Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Single Cell and Spatial Transcriptomics Define a Proinflammatory and Profibrotic Niche After Kidney Injury"

Article Title: Single Cell and Spatial Transcriptomics Define a Proinflammatory and Profibrotic Niche After Kidney Injury

Journal: Advanced Science

doi: 10.1002/advs.202503691

Figure Legend Snippet: Single‐cell and spatial transcriptome landscape of healthy and fibrotic kidneys after unilateral ischemia‐reperfusion injury (UIRI). a) Schematic representation of single‐cell RNA sequencing (scRNA‐seq) and spatial transcriptomics (ST) of kidneys from the sham and 10‐day UIRI mice, graphically designed with Biorender ( https://www.biorender.com/ ). b) t‐SNE plot illustrating the intricate cellular diversity in fibrotic kidneys, demonstrating distinct clusters representing glomerular endothelial cells (GEC), podocytes (Podo), mesangial cells (Mesa), Bowman's capsule epithelium (BC), proximal tubules (PT), descending limbs of Henle (DLOH), ascending limbs of Henle (ALOH), distal tubules (DT), principal cells (PC), intercalated cells (IC), fibroblasts (Fib), smooth muscle cells (SMC), extraglomerular endothelial cells (EGEC), monocytes (Mono), dendritic cells (DC), macrophages (Mϕ), plasmacytoid dendritic cells (pDC), proliferating mononuclear lineage (Prolif mono_L), and neutrophils (Neu), B cells (B), T cells (T), proliferating T cells (prolif T), and natural killer cells (NK). These cell types were further categorized into four major compartments: Glomerular, Renal, Interstitium, and Immune, as indicated by color grouping in the plot. c) Bubble plot illustrating the relative proportions of major kidney cell types in sham and UIRI samples. Each dot represents the proportion of a given cell type in a specific sample group, with dot size corresponding to its relative proportion. d) A comprehensive heatmap depicting the unique marker gene signature of major renal cell types. e) UMAP plot illustrating the inferred renal cell region distribution based on integrated spatial transcriptomics data from normal (Sham) and UIRI 10D mouse kidneys, generated using the 10x Genomics Visium platform. The identified regions include glomerular cells (Glom), distinct segments of the proximal tubule (PTS1, PTS1S2, PTS2), injured proximal tubules (InjPT), ascending limbs of Henle in cortex (ALOH(C)), distal tubules (DT), connecting tubules and collecting ducts (CNT_CD), cells at the corticomedullary junction (CMJ), fibrogenic niche regions (Niche1, Niche2), the inner stripe of the outer medulla (IOM), inner medulla (IM), renal capsule (RC), and perirenal tissue (Perirenal). f) Spatial maps illustrating the anatomical distribution of renal cell regions in Sham and UIRI 10D mouse kidneys. Region colors correspond to the classifications defined in panel (e). g) Bubble plot illustrating the relative proportions of major renal cell regions in spatial transcriptomics data from sham and UIRI 10D mouse kidneys. h) Bubble plot depicting the expression patterns of marker genes across distinct renal cell regions in spatial transcriptomics data. Dot color indicates the average gene expression level within each region, while dot size represents the proportion of spatial spots expressing the gene. i) Schematic diagram of nephron segmentation by cell types. j) Comparison of kidney anatomical regions and spatial transcriptomic clusters, showing clusters in kidney tissue (top) and the corresponding Visium H&E‐stained section (bottom). k) Renal tissue structure alterations at the corticomedullary junction (CMJ) in UIRI samples, showing the formation of two distinct fibrogenic niches, Niche1 and Niche2. l) A heatmap showing the deconvolution scores of cell type compositions across different regions in Visium spatial transcriptomics data, obtained using the RCTD method. m) Spatial FeaturePlots of RCTD‐derived cell type scores in the sham (top) and UIRI (bottom) groups, with paired panels sharing a common legend.

Techniques Used: RNA Sequencing, Marker, Generated, Expressing, Gene Expression, Comparison, Staining, Derivative Assay

High‐resolution spatial transcriptomics and immunostaining reveal the TNC‐enriched fibroblast‐macrophage niche organization in fibrotic kidneys. a) Schematic diagram of the Visium HD workflow applied to kidney tissues from sham and UIRI model mice. b) UMAP visualization of integrated Visium HD spatial transcriptomics data from control mice (obtained from the 10x Genomics public dataset) and UIRI mice (this study), processed using canonical correlation analysis (CCA). This dimensionality reduction visualization reveals distinct clusters representing various renal parenchymal and stromal cell populations, including: Glomerulus, Vasculature, PTS1, PTS2, PTS1S2, InjPT, ascending limbs of Henle in cortex [ALOH(Cortex)], distal tubule and connecting tubule (DT_CNT), connecting tubule and collecting duct (CNT_CD), collecting duct in cortex [CD(Cortex)], PTS3, injured PTS3 (InjPTS3), Fibrogenic Niche, Vasa recta, loop of Henle in outer medulla [LOH(IOM)], collecting duct in outer medulla [CD(IOM)], collecting duct in inner medulla [CD(IM)], thin ascending limbs of Henle in inner medulla [tALOH(IM)], renal capsule (RC), Perirenal Fibrous tissue, and Perirenal Adipose tissue. c) Bubble plot comparing the regional distribution in Control versus UIRI 10d kidneys (Visium HD). d) Bubble plot depicting the expression patterns of marker genes across distinct renal cell regions in Visium HD data. e) Spatial maps generated using Visium HD illustrate the inferred anatomical distribution of renal cell regions in kidney tissues from Control and UIRI mice. f) Spatial Feature Plots of Visium HD data showing the spatial distribution of selected renal cell types in controls (top) and UIRI mice (bottom), based on cell‐type deconvolution using RCTD. g) A heatmap showing the correlation between NMF factors and cell‐type deconvolution scores in standard Visium spatial transcriptomics data. h) Spatial distribution of gene scores associated with the NMF factors most correlated with the fibrogenic niche, along with the contribution of key genes to each factor. i) Spatial FeaturePlots showing the anatomical distribution of Tnc expression in standard Visium. j) A heatmap showing the correlation between NMF factors and cell type deconvolution scores in Visium HD spatial transcriptomics data. k) Spatial distribution of NMF factors (NMF3 and NMF11) associated with the fibrogenic niche in Visium HD data, along with their corresponding high‐contributing genes. l) Spatial FeaturePlots showing the anatomical distribution of Tnc expression in Visium HD datasets. m) Immunofluorescence staining demonstrates colocalization of TNC with macrophages (F4/80⁺) in the CMJ interstitial region. From top to bottom: an overview merged image (Merge), followed by magnified views of TNC, Vimentin, and F4/80 staining in the same region, and an enlarged merged image (Enlarged Merge) at the bottom.

Figure Legend Snippet: High‐resolution spatial transcriptomics and immunostaining reveal the TNC‐enriched fibroblast‐macrophage niche organization in fibrotic kidneys. a) Schematic diagram of the Visium HD workflow applied to kidney tissues from sham and UIRI model mice. b) UMAP visualization of integrated Visium HD spatial transcriptomics data from control mice (obtained from the 10x Genomics public dataset) and UIRI mice (this study), processed using canonical correlation analysis (CCA). This dimensionality reduction visualization reveals distinct clusters representing various renal parenchymal and stromal cell populations, including: Glomerulus, Vasculature, PTS1, PTS2, PTS1S2, InjPT, ascending limbs of Henle in cortex [ALOH(Cortex)], distal tubule and connecting tubule (DT_CNT), connecting tubule and collecting duct (CNT_CD), collecting duct in cortex [CD(Cortex)], PTS3, injured PTS3 (InjPTS3), Fibrogenic Niche, Vasa recta, loop of Henle in outer medulla [LOH(IOM)], collecting duct in outer medulla [CD(IOM)], collecting duct in inner medulla [CD(IM)], thin ascending limbs of Henle in inner medulla [tALOH(IM)], renal capsule (RC), Perirenal Fibrous tissue, and Perirenal Adipose tissue. c) Bubble plot comparing the regional distribution in Control versus UIRI 10d kidneys (Visium HD). d) Bubble plot depicting the expression patterns of marker genes across distinct renal cell regions in Visium HD data. e) Spatial maps generated using Visium HD illustrate the inferred anatomical distribution of renal cell regions in kidney tissues from Control and UIRI mice. f) Spatial Feature Plots of Visium HD data showing the spatial distribution of selected renal cell types in controls (top) and UIRI mice (bottom), based on cell‐type deconvolution using RCTD. g) A heatmap showing the correlation between NMF factors and cell‐type deconvolution scores in standard Visium spatial transcriptomics data. h) Spatial distribution of gene scores associated with the NMF factors most correlated with the fibrogenic niche, along with the contribution of key genes to each factor. i) Spatial FeaturePlots showing the anatomical distribution of Tnc expression in standard Visium. j) A heatmap showing the correlation between NMF factors and cell type deconvolution scores in Visium HD spatial transcriptomics data. k) Spatial distribution of NMF factors (NMF3 and NMF11) associated with the fibrogenic niche in Visium HD data, along with their corresponding high‐contributing genes. l) Spatial FeaturePlots showing the anatomical distribution of Tnc expression in Visium HD datasets. m) Immunofluorescence staining demonstrates colocalization of TNC with macrophages (F4/80⁺) in the CMJ interstitial region. From top to bottom: an overview merged image (Merge), followed by magnified views of TNC, Vimentin, and F4/80 staining in the same region, and an enlarged merged image (Enlarged Merge) at the bottom.

Techniques Used: Immunostaining, Control, Expressing, Marker, Generated, Immunofluorescence, Staining

TLR4 knockout in macrophages attenuates renal inflammation and renal fibrosis in vivo. a) The diagram shows the experimental protocol. Bone marrow chimera models were established by transplanting the WT bone marrow to WT mice, or TLR4 KO bone marrow to WT mice. Mice were irradiated at a single dose of 1100 Rads and then underwent bone marrow transplantation. After 8 weeks of successful transplantation, a unilateral ischemia‐reperfusion (UIRI) model was established. b) PCR‐based identification of kidney genotypes in the recipient mice of bone marrow transplantation models using TLR4 mutation site primers and wild‐type site primers, respectively. c,d) Graphic presentations show serum creatinine (Scr) (c) and blood urea nitrogen (BUN) (d) levels in different groups as indicated at 11 days after IRI. * p < 0.05 versus WT‐WT (n = 4–6). e,f) Western blot analyses show renal expression of TLR4, p‐P65, and P65 in different groups as indicated. Representative Western blot (e) and quantitative data (f) are shown. * p < 0.05 versus WT‐WT (n = 4–6). g) Representative micrographs show renal expression and co‐localization of TLR4 and F4/80 by immunofluorescence staining in different groups as indicated. The areas between the dashed lines represent the corticomedullary junction of the kidney. h,i) Western blot analyses show renal expression of MR, Arg‐1, iNOS, TNF‐α, and CCL2 in different groups as indicated. Representative Western blot (h) and quantitative data (i) are shown. * p < 0.05 versus WT‐WT (n = 4–6). j,k) Western blot analyses show renal expression of TNC, FN, and α‐SMA in different groups as indicated. Representative Western blot (j) and quantitative data (k) are shown. * p < 0.05 versus WT‐WT (n = 4–6). l) A schematic diagram shows a crucial role of TNC in organizing the proinflammatory and profibrotic niche. By integrating single‐cell RNA sequencing and spatial transcriptomics, we unveil TNC as a central organizer of the proinflammatory and profibrotic niche in kidney fibrosis. TNC promotes macrophage activation through TLR4/NF‐κB signaling, leading to macrophage activation, proliferation, and cytokine production.

Figure Legend Snippet: TLR4 knockout in macrophages attenuates renal inflammation and renal fibrosis in vivo. a) The diagram shows the experimental protocol. Bone marrow chimera models were established by transplanting the WT bone marrow to WT mice, or TLR4 KO bone marrow to WT mice. Mice were irradiated at a single dose of 1100 Rads and then underwent bone marrow transplantation. After 8 weeks of successful transplantation, a unilateral ischemia‐reperfusion (UIRI) model was established. b) PCR‐based identification of kidney genotypes in the recipient mice of bone marrow transplantation models using TLR4 mutation site primers and wild‐type site primers, respectively. c,d) Graphic presentations show serum creatinine (Scr) (c) and blood urea nitrogen (BUN) (d) levels in different groups as indicated at 11 days after IRI. * p < 0.05 versus WT‐WT (n = 4–6). e,f) Western blot analyses show renal expression of TLR4, p‐P65, and P65 in different groups as indicated. Representative Western blot (e) and quantitative data (f) are shown. * p < 0.05 versus WT‐WT (n = 4–6). g) Representative micrographs show renal expression and co‐localization of TLR4 and F4/80 by immunofluorescence staining in different groups as indicated. The areas between the dashed lines represent the corticomedullary junction of the kidney. h,i) Western blot analyses show renal expression of MR, Arg‐1, iNOS, TNF‐α, and CCL2 in different groups as indicated. Representative Western blot (h) and quantitative data (i) are shown. * p < 0.05 versus WT‐WT (n = 4–6). j,k) Western blot analyses show renal expression of TNC, FN, and α‐SMA in different groups as indicated. Representative Western blot (j) and quantitative data (k) are shown. * p < 0.05 versus WT‐WT (n = 4–6). l) A schematic diagram shows a crucial role of TNC in organizing the proinflammatory and profibrotic niche. By integrating single‐cell RNA sequencing and spatial transcriptomics, we unveil TNC as a central organizer of the proinflammatory and profibrotic niche in kidney fibrosis. TNC promotes macrophage activation through TLR4/NF‐κB signaling, leading to macrophage activation, proliferation, and cytokine production.

Techniques Used: Knock-Out, In Vivo, Irradiation, Transplantation Assay, Mutagenesis, Western Blot, Expressing, Immunofluorescence, Staining, RNA Sequencing, Activation Assay

Spatial Transcriptomics:

Article Title: Single Cell and Spatial Transcriptomics Define a Proinflammatory and Profibrotic Niche After Kidney Injury
Article Snippet: .. For the preparation of sections for Visium Spatial Transcriptomics sequencing, samples were equilibrated at −18 °C and a 10 μm thick section was cut onto the active sequencing area (6 mm x 6 mm) of a spatial barcoded slide. ..

Sequencing:

Image Search Results

A HE-stained image of the Visium tissue section and adjacent Xenium section, alongside their alignment. One repeat is performed for the publicly available data. B Pearson correlation between FineST and iStar for all input genes, calculated after aggregating super-resolution data to spot resolution. C Spatial expression plots for OPRPN from left to right: Visium, Xenium, FineST and iStar. FineST enhances the signal relative to Visium and yields results more comparable to Xenium. D Pearson correlation for OPRPN in FineST, corresponding to panel ( C ). Each dot represents a Visium spot ( n = 4992) or overlapping Xenium pseudo-spot ( n = 3958). E Ground-truth cell type annotations at spot (Visium) and single-cell (Xenium) resolution, as reported previously . F FineST's predicted cell types at single-nucleus and sub-spot levels. G FineST accurately identifies the DCIS 2 cell type in a triple-positive receptor ROI. H Pearson correlation for cell type abundance and mean gene expression across each cell type, comparing FineST and iStar. Each dot represents a cell type (Left, n = 19) or gene (Right, n = 65), lines connect matched pairs. Diamond indicates the mean. Statistical significance was assessed by a paired two-sided t -test. I Three marked regions (ROI 1, ROI 2 and ROI 3) dominated by DCIS 1, DCIS 2 and Invasive tumor cells. J Cell type deconvolution from FineST, compared with Xenium ground truth, demonstrates FineST's results visually match the ground truth and outperform Visium’s lower resolution (see Supplementary Fig. ). Alongside, single-cell resolved CCC patterns identified using SparseAEH (cluster number 2) and pathway enrichment analysis for Pattern 0 correspond to interesting cell distributions. K Venn plot of significant LR pairs (FDR < 0.05) interacting in >25% (ROI 1, 5589 cells) or >20% (ROI 2, 3330 cells; ROI 3, 5853 cells) of cells. In total, 103, 146 and 159 pairs were selected for spatial clustering analysis in the three ROIs, respectively. L Comparative analysis of region- and cell-specific LR pairs reveals two unique pairs specific to DCIS 2 and Invasive tumor cells. For panels ( B ), ( H – J ), source data are provided in the Source Data file. Scare bars, 1 mm.

Journal: Nature Communications

Article Title: FineST: contrastive learning integrates histology and spatial transcriptomics for nuclei-resolved ligand-receptor analysis

doi: 10.1038/s41467-026-70528-7

Figure Lengend Snippet: A HE-stained image of the Visium tissue section and adjacent Xenium section, alongside their alignment. One repeat is performed for the publicly available data. B Pearson correlation between FineST and iStar for all input genes, calculated after aggregating super-resolution data to spot resolution. C Spatial expression plots for OPRPN from left to right: Visium, Xenium, FineST and iStar. FineST enhances the signal relative to Visium and yields results more comparable to Xenium. D Pearson correlation for OPRPN in FineST, corresponding to panel ( C ). Each dot represents a Visium spot ( n = 4992) or overlapping Xenium pseudo-spot ( n = 3958). E Ground-truth cell type annotations at spot (Visium) and single-cell (Xenium) resolution, as reported previously . F FineST's predicted cell types at single-nucleus and sub-spot levels. G FineST accurately identifies the DCIS 2 cell type in a triple-positive receptor ROI. H Pearson correlation for cell type abundance and mean gene expression across each cell type, comparing FineST and iStar. Each dot represents a cell type (Left, n = 19) or gene (Right, n = 65), lines connect matched pairs. Diamond indicates the mean. Statistical significance was assessed by a paired two-sided t -test. I Three marked regions (ROI 1, ROI 2 and ROI 3) dominated by DCIS 1, DCIS 2 and Invasive tumor cells. J Cell type deconvolution from FineST, compared with Xenium ground truth, demonstrates FineST's results visually match the ground truth and outperform Visium’s lower resolution (see Supplementary Fig. ). Alongside, single-cell resolved CCC patterns identified using SparseAEH (cluster number 2) and pathway enrichment analysis for Pattern 0 correspond to interesting cell distributions. K Venn plot of significant LR pairs (FDR < 0.05) interacting in >25% (ROI 1, 5589 cells) or >20% (ROI 2, 3330 cells; ROI 3, 5853 cells) of cells. In total, 103, 146 and 159 pairs were selected for spatial clustering analysis in the three ROIs, respectively. L Comparative analysis of region- and cell-specific LR pairs reveals two unique pairs specific to DCIS 2 and Invasive tumor cells. For panels ( B ), ( H – J ), source data are provided in the Source Data file. Scare bars, 1 mm.

Article Snippet: The raw and processed Visium spatial sequencing data of human hepatocellular carcinoma (HCC) tissues were downloaded from Mendeley Data under accession number skrx2fz79n [ https://data.mendeley.com/datasets/skrx2fz79n/1 ], while the cell type annotations and high-resolution HE-stained images were provided by the original corresponding author.

Techniques: Staining, Expressing, Single Cell, Gene Expression

$A , B Spatial plot of seven cell types estimated by cell2location (spot resolution) and FineST (single-cell resolution). A' , B' Zoon-in views of A and B . B'' Zoon-in view of the region marked in B' , where FineST increases resolution from 8 spots to 869 single cells. C Cell type composition from reference scRNA-seq and deconvolution at Visium spot and FineST single-cell levels. D Distribution and PCC of cell type proportions ( n = 7) for Visium spot (PCC = 0.24) and FineST single-cell (PCC = 0.64) resolutions vs reference scRNA-seq. E Pathologists identified 36 spots (of 1331) co-localized with tertiary lymphoid structure (TLS), important for antigen presentation and T cell activation. FineST's single-cell views validate TLS by T and B cell co-localization (see B'' ). F Three example spatially co-expressed LR pairs detected at FineST's single-cell resolution. G Visualization of PVR - TIGIT interaction among local single cells ( z -score FDR < 0.05). H Clustering of 633 significant LR pairs, from single-cell resolution, within selected ROI into three spatial patterns using SpatialDE. I Spatial co-localization of tumor and Treg cells, estimated by cell2location . J Spatially co-expressed LR pairs detected at FineST's single-cell resolution. Scatter plot of global Moran’s R and one-sided z -score p -values (orange: significant, FDR < 0.05, Benjamini-Hochberg correction), with examples highlighted. K Communication strength of CD70 - CD27 at single-cell resolution (color: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$1-{p}_{{{{\rm{local}}}}{z}_{p}}$$\end{document} 1 − p local z p ; mean strength: 0.45, interacting cells: 5961, occupancy: 49.2%). L Detection of local CCC patterns by clustering significant LR pairs. Top: original spot resolution (332 pairs, SpatialDE); Middle: sub-spot resolution (957 pairs, SparseAEH) and Bottom: single-nucleus resolution (931 pairs, SparseAEH). M Dot plots of enriched pathways in Pattern 0 and Pattern 1 (from L , Bottom), 2-by-2 contingency tables for MHC-I and WNT. Statistical significance was assessed using a one-sided Fisher’s exact test; dot size indicates p -value. N Sankey plot of selected L-R-TF-TG communication pathways. For panels (D , K ), box plots show median (center), IQR (box), whiskers at 1.5 × IQR, and points for seven cell types; violin plots show density, median (white line), and IQR (thick bar). For panels ( A ), ( J , L ), source data are provided in the Source Data file.$

Journal: Nature Communications

Article Title: FineST: contrastive learning integrates histology and spatial transcriptomics for nuclei-resolved ligand-receptor analysis

doi: 10.1038/s41467-026-70528-7

Figure Lengend Snippet: A , B Spatial plot of seven cell types estimated by cell2location (spot resolution) and FineST (single-cell resolution). A' , B' Zoon-in views of A and B . B'' Zoon-in view of the region marked in B' , where FineST increases resolution from 8 spots to 869 single cells. C Cell type composition from reference scRNA-seq and deconvolution at Visium spot and FineST single-cell levels. D Distribution and PCC of cell type proportions ( n = 7) for Visium spot (PCC = 0.24) and FineST single-cell (PCC = 0.64) resolutions vs reference scRNA-seq. E Pathologists identified 36 spots (of 1331) co-localized with tertiary lymphoid structure (TLS), important for antigen presentation and T cell activation. FineST's single-cell views validate TLS by T and B cell co-localization (see B'' ). F Three example spatially co-expressed LR pairs detected at FineST's single-cell resolution. G Visualization of PVR - TIGIT interaction among local single cells ( z -score FDR < 0.05). H Clustering of 633 significant LR pairs, from single-cell resolution, within selected ROI into three spatial patterns using SpatialDE. I Spatial co-localization of tumor and Treg cells, estimated by cell2location . J Spatially co-expressed LR pairs detected at FineST's single-cell resolution. Scatter plot of global Moran’s R and one-sided z -score p -values (orange: significant, FDR < 0.05, Benjamini-Hochberg correction), with examples highlighted. K Communication strength of CD70 - CD27 at single-cell resolution (color: \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$1-{p}_{{{{\rm{local}}}}{z}_{p}}$$\end{document} 1 − p local z p ; mean strength: 0.45, interacting cells: 5961, occupancy: 49.2%). L Detection of local CCC patterns by clustering significant LR pairs. Top: original spot resolution (332 pairs, SpatialDE); Middle: sub-spot resolution (957 pairs, SparseAEH) and Bottom: single-nucleus resolution (931 pairs, SparseAEH). M Dot plots of enriched pathways in Pattern 0 and Pattern 1 (from L , Bottom), 2-by-2 contingency tables for MHC-I and WNT. Statistical significance was assessed using a one-sided Fisher’s exact test; dot size indicates p -value. N Sankey plot of selected L-R-TF-TG communication pathways. For panels (D , K ), box plots show median (center), IQR (box), whiskers at 1.5 × IQR, and points for seven cell types; violin plots show density, median (white line), and IQR (thick bar). For panels ( A ), ( J , L ), source data are provided in the Source Data file.

Techniques: Single Cell, Immunopeptidomics, Activation Assay

A , B HE staining and cell type annotation of spatial transcriptomic spots in tumor tissues from an ICB non-responder (P1_T) and responder (P7_T) from the original study . In the non-responder (P1_T), a tumor immune barrier (TIB) structure is formed by SPP1 + macrophages and cancer-associated fibroblasts (CAFs). C , D Spatial signature score of SPP1 + macrophages and CAFs, spatial gene expression of CCR1 , and Pearson correlation between SPP1 + macrophage score and CCR1 expression across all spots (Visium vs FineST). The red line represents the fitted linear regression, and the shaded area corresponds to the 95% confidence interval. Statistical significance was assessed using a two-sided Pearson correlation test. E Scatterplot of global Moran’s R and one-sided z -score p -values for spatially co-expressed LR pairs detected using FineST-enhanced gene expression at spot-level resolution in ROI. Significant pairs (orange) were identified using FDR < 0.05 (Benjamini-Hochberg correction). The CD274-PDCD1 interaction was detected in P1_T only. F Spatial gene expression of ligand CD274 and receptor PDCD1 across all spots (Visium vs FineST) for P1_T (non-responder) and P7_T (responder), respectively. For panels ( A , B , E ), source data are provided in the Source Data file. Part of the panels ( A , B ) is created in BioRender. Huang, Y. (2026) https://BioRender.com/fv0byvk .

Journal: Nature Communications

Article Title: FineST: contrastive learning integrates histology and spatial transcriptomics for nuclei-resolved ligand-receptor analysis

doi: 10.1038/s41467-026-70528-7

Figure Lengend Snippet: A , B HE staining and cell type annotation of spatial transcriptomic spots in tumor tissues from an ICB non-responder (P1_T) and responder (P7_T) from the original study . In the non-responder (P1_T), a tumor immune barrier (TIB) structure is formed by SPP1 + macrophages and cancer-associated fibroblasts (CAFs). C , D Spatial signature score of SPP1 + macrophages and CAFs, spatial gene expression of CCR1 , and Pearson correlation between SPP1 + macrophage score and CCR1 expression across all spots (Visium vs FineST). The red line represents the fitted linear regression, and the shaded area corresponds to the 95% confidence interval. Statistical significance was assessed using a two-sided Pearson correlation test. E Scatterplot of global Moran’s R and one-sided z -score p -values for spatially co-expressed LR pairs detected using FineST-enhanced gene expression at spot-level resolution in ROI. Significant pairs (orange) were identified using FDR < 0.05 (Benjamini-Hochberg correction). The CD274-PDCD1 interaction was detected in P1_T only. F Spatial gene expression of ligand CD274 and receptor PDCD1 across all spots (Visium vs FineST) for P1_T (non-responder) and P7_T (responder), respectively. For panels ( A , B , E ), source data are provided in the Source Data file. Part of the panels ( A , B ) is created in BioRender. Huang, Y. (2026) https://BioRender.com/fv0byvk .

Techniques: Staining, Gene Expression, Expressing

Spatial Transcriptomics in horizontally sectioned E14.5 mouse diaphragm identifies distinct tissues and muscle domains (A) Schematic representation of embryonic mouse diaphragm in which relevant anatomical regions are indicated. NMJ: Neuromuscular junction, MTJ: Myotendinous junction. (B) SpatialFeaturePlots demonstrating expression levels and distributions of representative genes for muscle, NMJ, MTJ and tendon. (C) SpatialDimPlot demonstrating the distribution of all the identified clusters within the diaphragm tissue. (D) Spatial distribution of representative Seurat clusters differentiating genetically specific domains in developing mouse diaphragm. (E) Uniform Manifold Approximation and Projection (UMAP) diagram of identified clusters of spatial RNA sequencing with muscle and endothelial clusters at the left and tendon and erythrocyte clusters at right. (F) FeaturePlots showing expression of muscle ( Ttn , Myh3 , Myh8 ), crural diaphragm ( Crlf1 ), NMJ ( Chrna1 , Chrng , Musk, Etv5 ), MTJ ( Col22a1 , Ankrd1 , Rxrg , Csrp3 ) and tendon ( Col12a1 , Antxr1, Tnmd , Tnc ) markers displayed by UMAP.

Journal: iScience

Article Title: Spatial transcriptomics in embryonic mouse diaphragm muscle reveals regional gradients and subdomains of developmental gene expression

doi: 10.1016/j.isci.2024.110018

Figure Lengend Snippet: Spatial Transcriptomics in horizontally sectioned E14.5 mouse diaphragm identifies distinct tissues and muscle domains (A) Schematic representation of embryonic mouse diaphragm in which relevant anatomical regions are indicated. NMJ: Neuromuscular junction, MTJ: Myotendinous junction. (B) SpatialFeaturePlots demonstrating expression levels and distributions of representative genes for muscle, NMJ, MTJ and tendon. (C) SpatialDimPlot demonstrating the distribution of all the identified clusters within the diaphragm tissue. (D) Spatial distribution of representative Seurat clusters differentiating genetically specific domains in developing mouse diaphragm. (E) Uniform Manifold Approximation and Projection (UMAP) diagram of identified clusters of spatial RNA sequencing with muscle and endothelial clusters at the left and tendon and erythrocyte clusters at right. (F) FeaturePlots showing expression of muscle ( Ttn , Myh3 , Myh8 ), crural diaphragm ( Crlf1 ), NMJ ( Chrna1 , Chrng , Musk, Etv5 ), MTJ ( Col22a1 , Ankrd1 , Rxrg , Csrp3 ) and tendon ( Col12a1 , Antxr1, Tnmd , Tnc ) markers displayed by UMAP.

Article Snippet: Visium Spatial Transcriptomics sequencing data were aligned using the default SpaceRanger (2.0.1) pipeline for FFPE slides in a Singularity Container running Ubuntu 22.04 on a high-performance cluster (Medical University of Innsbruck).

Techniques: Expressing, RNA Sequencing

Spatial Transcriptomics reveals distinct myogenic processes in the muscle center and periphery in E14.5 mouse diaphragm (A) SpatialFeaturePlots demonstrating expression levels and distributions of myogenic differentiation markers. (B) VlnPlots of expression levels of genes in clusters identified as muscle center, default muscle, and muscle periphery show an overall increase or decrease in expression of genes involved in muscle development from the center to the periphery. Y axis indicates expression levels. (C) Cnetplots showing GO terms in biological processes (red nodes) and their associated genes (blue nodes) for the upregulated genes in clusters annotated as muscle center compared to muscle periphery (top) or muscle periphery compared to muscle center (bottom). (D) Dotplots of expression of genes involved in myogenesis displaying a declining (right) or increasing (left) gradient in clusters from the muscle center, over default muscle (muscle middle), to the muscle periphery.

Journal: iScience

Article Title: Spatial transcriptomics in embryonic mouse diaphragm muscle reveals regional gradients and subdomains of developmental gene expression

doi: 10.1016/j.isci.2024.110018

Figure Lengend Snippet: Spatial Transcriptomics reveals distinct myogenic processes in the muscle center and periphery in E14.5 mouse diaphragm (A) SpatialFeaturePlots demonstrating expression levels and distributions of myogenic differentiation markers. (B) VlnPlots of expression levels of genes in clusters identified as muscle center, default muscle, and muscle periphery show an overall increase or decrease in expression of genes involved in muscle development from the center to the periphery. Y axis indicates expression levels. (C) Cnetplots showing GO terms in biological processes (red nodes) and their associated genes (blue nodes) for the upregulated genes in clusters annotated as muscle center compared to muscle periphery (top) or muscle periphery compared to muscle center (bottom). (D) Dotplots of expression of genes involved in myogenesis displaying a declining (right) or increasing (left) gradient in clusters from the muscle center, over default muscle (muscle middle), to the muscle periphery.

Techniques: Expressing

Spatial Transcriptomics in horizontally sectioned E18.5 mouse diaphragm identifies specific functional muscle domains and fiber types (A) SpatialFeaturePlots demonstrating expression levels and distributions of NMJ genes ( Chrna1 , Musk , Etv5 , Ache , Chrng, Chrne ), neonatal ( Myh8 ) and embryonic ( Myh3 ) myosin heavy chains, developmental troponin Tnnt2 , and ventral diaphragm markers ( Myog , Flnc , Csrp3 ). (B) Spatial distribution of Seurat clusters of distinct muscle and tendon domains. (C) UMAP representation of identified clusters of spatial RNA sequencing indicates spatially and functionally divergent differentiation of diaphragm muscle. (D) FeaturePlots demonstrating expression of muscle ( Ttn , Myh3 , Myh8 ), type I muscle ( Myh7, Myl2, Myl3 ), type IIb muscle ( Myh4 , Pvalb , Mybpc2 ), NMJ ( Chrna1 , Musk , Etv5 ), MTJ ( Col22a1 , Ankrd1 , Uchl1 ) and tendon ( Tnmd , Col11a1 , Scx ) markers displayed by UMAP.

Journal: iScience

Article Title: Spatial transcriptomics in embryonic mouse diaphragm muscle reveals regional gradients and subdomains of developmental gene expression

doi: 10.1016/j.isci.2024.110018

Figure Lengend Snippet: Spatial Transcriptomics in horizontally sectioned E18.5 mouse diaphragm identifies specific functional muscle domains and fiber types (A) SpatialFeaturePlots demonstrating expression levels and distributions of NMJ genes ( Chrna1 , Musk , Etv5 , Ache , Chrng, Chrne ), neonatal ( Myh8 ) and embryonic ( Myh3 ) myosin heavy chains, developmental troponin Tnnt2 , and ventral diaphragm markers ( Myog , Flnc , Csrp3 ). (B) Spatial distribution of Seurat clusters of distinct muscle and tendon domains. (C) UMAP representation of identified clusters of spatial RNA sequencing indicates spatially and functionally divergent differentiation of diaphragm muscle. (D) FeaturePlots demonstrating expression of muscle ( Ttn , Myh3 , Myh8 ), type I muscle ( Myh7, Myl2, Myl3 ), type IIb muscle ( Myh4 , Pvalb , Mybpc2 ), NMJ ( Chrna1 , Musk , Etv5 ), MTJ ( Col22a1 , Ankrd1 , Uchl1 ) and tendon ( Tnmd , Col11a1 , Scx ) markers displayed by UMAP.

Techniques: Functional Assay, Expressing, RNA Sequencing

Spatial transcriptomics reveals aberrant regulation of myogenic genes in Ca V 1.1 −/− mice (A) FeaturePlots showing expression of Ttn (muscle), Chrna1 and Musk (NMJ) displayed by UMAP in control and Ca V 1.1 −/− integrated dataset at E14.5 (left) and E18.5 (right). (B) Violin plots showing expression of representative genes differentially expressed in control and Ca V 1.1 −/− samples at E14.5 and E18.5. Y axis indicates expression level. (C) FeaturePlots of module scores of muscle differentiation markers displayed by UMAP in E18.5 control and Ca V 1.1 −/− integrated dataset show increased expression of early markers (top) and a decreased expression of late markers (bottom) in Ca V 1.1 −/− muscles. (D) Venn diagrams of top 200 DEGs genes and GO terms for these genes between E14.5 control and E18.5 control and between E18.5 Ca V 1.1 −/− and E18.5 control muscles indicate more shared genes and GO terms for upregulated genes in E18.5 Ca V 1.1 −/− with E14.5 control muscle and for downregulated genes in E18.5 Ca V 1.1 −/− with E18.5 control muscle. (E) FeaturePlots showing expression of Klf5 and Tead4 displayed by UMAP in control and Ca V 1.1 −/− integrated dataset at E14.5 (left) and E18.5 (right). (F) Violin plots of Klf5 and Tead4 expression in muscle clusters of E14.5 and E18.5 control and Ca V 1.1 −/− spatial datasets. Y axis indicates expression level.

Journal: iScience

Article Title: Spatial transcriptomics in embryonic mouse diaphragm muscle reveals regional gradients and subdomains of developmental gene expression

doi: 10.1016/j.isci.2024.110018

Figure Lengend Snippet: Spatial transcriptomics reveals aberrant regulation of myogenic genes in Ca V 1.1 −/− mice (A) FeaturePlots showing expression of Ttn (muscle), Chrna1 and Musk (NMJ) displayed by UMAP in control and Ca V 1.1 −/− integrated dataset at E14.5 (left) and E18.5 (right). (B) Violin plots showing expression of representative genes differentially expressed in control and Ca V 1.1 −/− samples at E14.5 and E18.5. Y axis indicates expression level. (C) FeaturePlots of module scores of muscle differentiation markers displayed by UMAP in E18.5 control and Ca V 1.1 −/− integrated dataset show increased expression of early markers (top) and a decreased expression of late markers (bottom) in Ca V 1.1 −/− muscles. (D) Venn diagrams of top 200 DEGs genes and GO terms for these genes between E14.5 control and E18.5 control and between E18.5 Ca V 1.1 −/− and E18.5 control muscles indicate more shared genes and GO terms for upregulated genes in E18.5 Ca V 1.1 −/− with E14.5 control muscle and for downregulated genes in E18.5 Ca V 1.1 −/− with E18.5 control muscle. (E) FeaturePlots showing expression of Klf5 and Tead4 displayed by UMAP in control and Ca V 1.1 −/− integrated dataset at E14.5 (left) and E18.5 (right). (F) Violin plots of Klf5 and Tead4 expression in muscle clusters of E14.5 and E18.5 control and Ca V 1.1 −/− spatial datasets. Y axis indicates expression level.

Techniques: Expressing, Control, Muscles

Review

visium spatial transcriptomics sequencing (Spatial Transcriptomics Inc)

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1) Product Images from "Single Cell and Spatial Transcriptomics Define a Proinflammatory and Profibrotic Niche After Kidney Injury"

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