Journal: bioRxiv
Article Title: Single-Cell and Spatial Methods for Multimodal Functional Glycan Profiling in Tissues
doi: 10.64898/2026.02.23.707224
Figure Lengend Snippet: (A) Schematic workflow of GlycoScope, illustrating the sequential staining of antibody and recombinant lectin panels. (B) Representative multiplexed images showing nuclei (DAPI), Siglec-9L, Siglec-15L, Galectin-8L, T cells (CD3), B cells (CD20), and follicular dendritic cells (CD21) across GlycoScope, Antibody-only, and Lectin-only conditions of tonsil tissues (scale bar: 50µm, 10µm (inset)). All individual images across these three conditions are in . (C) Heatmap showing the 0,1 normalization of mean expression (left) and z-score of the coefficient of variation (right) of all glycan and protein markers obtained from GlycoScope, Antibody-only, and Lectin-only conditions. (D) Schematic overview of the GlycoScope study cohorts. Cohort 1 consists of an FFPE tissue microarray (TMA) containing four tonsil, four follicular hyperplasia (FH), and eight follicular lymphoma (FL) cores, profiled using a 32-plex protein panel and six oligo-conjugated lectins (Siglec-7, Siglec-9, Siglec-15, MBL, DC-SIGN, and Galectin-8). Cohort 2 consists of an FFPE TMA containing 11 FL and 5 FH samples, profiled using a 42-plex protein panel and six oligo-conjugated lectins (Siglec-7, Siglec-9, Siglec-15, DC-SIGN, Galectin-8, and Galectin-9). (E) Representative GlycoScope multiplexed images (top) showing nuclei (DAPI), B/tumor cells (Pax5), macro-phages (CD68), T cells (CD3), endothelial cells (CD31), FDC-enriched region (CD21) and lymphatic (Podoplanin), as well as the corresponding phenotype maps (bottom) of FL, FH and tonsil tissues (scale bar: 100µm, 20µm (inset)). All individual phenotype maps for each tissue sample core are in . (F) Top: Representative GlycoScope multiplexed images of FL and FH showing Siglec-7L (magenta), CD31 (yellow), and PAX5 (cyan). Bottom: Representative GlycoScope multiplexed images of FL and FH showing Siglec-9L (cyan), CD31 (magenta), and PAX5 (white). Scale bar, 100 µm. (G) Representative GlycoScope multiplexed images of FL and FH showing DC-SIGNL (cyan), CD21 (magenta) and CD21 (yellow). scale bar: 100µm. (H) Violin plots showing the expression of DC-SIGNL acquired from GlycoScope in tonsil, FH and FL tissues (Krus-kal-Wallis rank sum test followed by pairwise Wilcoxon test with Benjamini-Hochberg adjustments). (I) Histogram showing the expression distribution of DC-SIGNL in B cell. (J) Violin plots showing the expression of BCL2 protein in DC-SIGNL High and DC-SIGNL Low B cells (Pairwise Wilcoxon test). (K) Line plot showing the mean expression of DC-SIGNL in follicle B cell at 0-150 µm from the anchored FDC. ***P<0.001; ****P<0.0001
Article Snippet: We defined the follicular center as the region located 80-150 μm inward from the Galectin-8L High mantle zone rim, the mantle zone as spanning 0-80 μm, and the marginal zone as extending outward of the rim.
Techniques: Staining, Recombinant, Expressing, Glycoproteomics, Microarray