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    Classification:

    Long-read Spatial Transcriptomics


    Introduction


    Spatial transcriptomics involves the use of Nanopore long-read sequencing technology to directly obtain data from Stereo-seq or 10X Visium HD 3', which provides spatially resolved information about full-length cDNA. By analyzing each spot, one can determine the results of isoform quantification and variable splicing events. It not only enables the identification of all RNA molecules but also retains the original positioning information within the tissue section. This is crucial for understanding the spatial distribution of gene expression, as the relationships within and between cells are significant.

    Applications


    Tumor research

    Brain science research

    Embryonic development

    Immunology

    Highlights


    Spatially specific isoforms

    Differential isoform variation

    Isoform exchange in different regions

    Workflow


    Sample Preparation

    Sample Preparation

    Refer to the Sample
    Submission Requirements

    Sectioning and imaging

    Sectioning and imaging

    Section thickness: OCT 10 μm
    Nucleic acid extraction and quality control from 20 sections

    Sample QC

    Sectioning and imaging
    Spatial Library Preparation

    Spatial Library Preparation

    · Tissue permeabilization, mRNA release and barcoding
    · cDNA synthesis & amplification
    · Long-read library construction

    Library QC

    Spatial Library Preparation
    Sequencing

    Sequencing

    Platform: Nanopore
    Strategy: 2-4 flow cells per sample
    Data volume: 100 G per flow cell

    Data QC

    Sequencing
    Bioinformatics Analysis

    Bioinformatics Analysis

    Refer to the demo results

    Click >>

    Analysis Workflow


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    Data Delivery File Formats


    Fastq Data Files

    Sequencing Yield Display


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    Sample Requirements


    Sample Type

    Platform

    Sample Collection

    Transportation

    OTC

    Visium HD

    Ensure that fresh tissues are embedded in OCT within 30 minutes after ex vivo isolation.

    Length × Width: ≤ 6.5 mm × 6.5 mm; Height: ≥ 5 mm

    Mark the cut surface with an asterisk (*) or the label "Cut Surface".

    Stored at -80℃.

    Pack the sorted samples and embed them in dry ice and send it out as soon as possible.

    Ensure sufficient dry ice is used.

    Stereo-seq

    Ensure that fresh tissues are embedded in OCT within 30 minutes after ex vivo isolation.

    The maximum size of the tissue cut surface shall not exceed 0.9 times the chip size. The thickness of the embedded block shall be controlled within 2 cm.

    Mark the cut surface with an asterisk (*) or the label "Cut Surface".

    Stored at -80℃.

    FFPE

    Visium HD

    The recommended thickness of paraffin blocks is 2–5 mm.

    Length × Width: ≤ 6.5 mm × 6.5 mm; Height: ≥ 5 mm

    Mark the cut surface with an asterisk (*) or the label "Cut Surface".

    Store at room temperature or 4℃.

    Fix the samples in sample boxes to avoid squeezing the surfaces of paraffin blocks or glass slides, and transport them at 4℃ or room temperature.

    Stereo-seq

    The maximum size of the tissue cut surface shall not exceed 0.9 times the chip size. The thickness of the embedded block shall be controlled within 2 cm.

    Mark the cut surface with an asterisk (*) or the label "Cut Surface".

    Store at room temperature or 4℃.

    Publications


    Year Journal

    Impact Factor

    Title
    2025 Nature Communications 15.7  A spatial long-read approach at near-singlecell resolution reveals developmental regulation of splicing and polyadenylation sites in distinct cortical layers and cell types
    2025 bioRxiv / Spatial isoform sequencing at sub-micrometer single-cell resolution reveals novel patterns of spatial isoform variability in brain cell types.
    2023 Nucleic Acids Research 13.1  The spatial landscape of gene expression isoforms in tissue sections.
    2023 Nature Communications 15.7  Spatial transcriptomics reveals niche-specific enrichment and vulnerabilities of radial glial stem-like cells in malignant gliomas.
    2021 Nature Communications 15.7  A spatially resolved brain region- and cell typespecific isoform atlas of the postnatal mouse brain.

    What's the best choice?


    Year

    Long-read spatial transcriptome sequencin

    Spatial transcriptome sequencing

    Sequencing Technology

    Nanopore PromethION

    Illumina / BGI

    Reads Length

    Mean read length ~600 bp

    150bp

    Delivery File Format

    Fastq

    Fastq

    Analyze Content

    1.Spatial gene expression, spatial clustering, spatial annotation

    2.Identification of isoform diversity

    3.Isoform quantification

    4.Differential transcript analysis for spatial clustering

    5.Comparison of gene clustering results with isoform clustering results

    6.Identification of spatial subpopulations through differential alternative splicing

    7.Isoform switching across spatial locations

    8.Identification of fusion genes in spatial regions

    9.Identification of lncRNAs in spatial regions

    1.Spatial gene expression, spatial clustering, spatial annotation

     


    Keyword:

    Long-read Spatial Transcriptomics


    Contact Us

    If you are interested in our long-read sequencing services or potential collaboration, please contact us. Our team is ready to support your research with tailored solutions. We also welcome feedback from users to help us improve our services.

    Contact Us
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    Email:service@sailgene.com

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