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Visium FFPE Spatial Transcriptome

Break through the barriers that once hindered spatial analysis of gene expression in formalin-fixed paraffin-embedded (FFPE) tissue sections and unveil the hidden insights within your samples with Visium Spatial Gene Expression for FFPE. By merging the advantages of histological techniques with the vast throughput and investigative capabilities of RNA sequencing in FFPE tissue samples, Visium Spatial Gene Expression for FFPE enhances traditional pathologist-led analysis. Profile RNA expression across entire tissue sections with high resolution, encompassing more than 18,000 genes in human and mouse FFPE samples, to gain spatial insight into gene expression patterns.

Visium HD empowers a new era of spatial discovery, enhancing proven whole transcriptome spatial analysis with single cell–scale resolution, enabling continuous tissue coverage, and delivering best-in-class data with innovative probe-based chemistry and a Visium CytAssist-enabled workflow.

Sample Requirements

Sample Type Sample Amount Preservation Sample QC Shipping
FFPE block
(recommended)
1 block, FFPE block must be
contained on plastic dehydrating box,
otherwise could not be installed on the
sectioning device
After embedding
store at 4°C,
protected from light
DV200% ≥ 30% 4°C or Room
Temperature
FFPE slide 5-10 FFPE (10μm thickness) scrolls in
tube for sample QC;

2-4 FFPE (5μm thickness) tissue
sections on glass slides, for library prep*
Dry and sealed,
storage time at 4°C
< 14 days
DV200% ≥ 30% 4°C or Room
Temperature

*FFPE glass slides must be positive charged glass slides.

Specifications: Sequencing and Analysis

  Visium HD
Species Human or mouse
Resolution 2 um (single cell level)
Sequencing platform NovaSeq X plus
Read length PE150
Recommended Data Amount Coverage Area x 275,000,000 read pairs~ 100 Gb/sample
Analysis
  • Data QC
  • Mapping and Quantification
  • Dimensionality Reduction, Clustering, and Differential analysis:
    • Graphclust clustering
    • K-means clustering
  • Seurat Analysis:
    • PCA analysis
  • Spatial variable gene expression analysis
    • Clustering and dimensionality reduction analysis
  • Differential Expression Gene Analysis (DEG)
    • DEGs clustering heatmap
    • Spatial distribution of DEGs
    • Violine plot
    • t-SNE and UMAP dimensional reduction visualization
  • Functional Enrichment Analysis:
    • GO enrichment
    • KEGG enrichment
    • Reactome enrichment

Project Workflow

Demo Results of Bioinformatics Analysis

Contact us for a demo report.

Figure 1. Spatial quantification of gene expression in bins

Space Ranger is used to count the UMI for each gene on every bin, obtaining a bin-gene expression matrix for the sample. After performing data QC, the figure below shows the distribution of UMI per gene across the spatial regions of the tissue.

Figure 2. Clustering analysis presentation: spatial distribution, tSNE and UMAP

Cluster_Spatial: spatial distribution map of clusters; Cluster_tSNE: tSNE two-dimensional spatial representation map of clusters; Cluster_UMAP: UMAP two-dimensional spatial representation map of clusters

Figure 3. t-SNE and UMAP dimensionality reduction of differentially expressed genes

The top 4 differentially expressed genes of each cluster are displayed in several ways. The figure presents the t-SNE and UMAP dimensional reduction visualization of differentially expressed genes.