Description

Single-Cell Nucleus Transcriptome Sequencing (Single Nuclei RNA Sequencing, snRNA-seq) involves extracting nuclei from samples and using the 10x Genomics platform to isolate and label these nuclei. This technology allows for the study of nuclear gene expression at the single-cell level. It provides a platform for single-cell research in complex tissues such as brain, heart, and kidney, as well as in rare frozen samples. This approach enables the exploration of potential pathogenic cell types and facilitates the investigation of tumor cell heterogeneity and disease mechanisms.

Principles

Single-cell suspension or tissue-derived single-cell nuclei: Nuclei are extracted, cleaned, and resuspended using our optimized nuclear isolation kit. The 10x Genomics single-cell sequencing platform then employs techniques such as microfluidics, droplet encapsulation, and barcode tagging to achieve high-throughput single-cell nucleus capture.

A single-cell nucleus transcriptome library is constructed, and sequencing is performed using the Illumina sequencing platform. This process allows for the simultaneous acquisition of gene expression data from a large number of single-cell nuclei, enabling gene expression sequencing at the single-cell level in complex tissues or precious frozen samples.

Advantages

• Simple Operation: Utilizes a direct nucleus extraction method, which is easier to perform as the nuclear membrane is more stable than the cell membrane.

• Wide Applicability: Suitable for fresh or frozen samples, difficult-to-digest tissues, and complex tissue types, with no limitations related to cell size or activity.

• Accurate Results: The process avoids introducing "transcriptional bias" during dissociation, ensuring reliable and authentic results.

• Comprehensive Information: Uniform tissue digestion yields accurate cell type composition and proportions, allowing for more complete and thorough cell type identification.

• High-Quality Nucleus Extraction: Optimized methods for preparing single-cell nucleus suspensions using the single-cell nucleus isolation kit for various tissues.

• Extensive Experience: Over 100 tissue types and 3,000+ single-cell samples prepared, with extensive experience in high-quality nucleus extraction.

• Powerful Bioinformatics Analysis: Enables visualization of single-nucleus data, uncovering rare cell types, and providing personalized analysis.

Applications

• Cell Atlas Construction: Mapping the diversity and characteristics of cell types within tissues.
• Growth and Development: Studying cellular processes and changes during growth and developmental stages.
• Neuroscience Research: Investigating neural cell types, brain function, and neurological disorders.
• Oncology Research: Exploring tumor heterogeneity, cancer cell types, and tumor microenvironment.
• Stem Cell Differentiation: Analyzing the differentiation processes of stem cells into various cell types.
• Discovery of Undetected Cell Types: Identifying previously unrecognized cell types within complex tissues.

Q&A

Question: Differences Between Single-Cell Nucleus and Single-Cell Transcriptome Sequencing

• Sample Preparation:

  • Single-Cell Nucleus RNA Sequencing: Requires the extraction and preparation of single-cell nucleus suspensions.
  • Single-Cell Transcriptome Sequencing: Only requires the preparation of single-cell suspensions. The genes analyzed are different, but both methods enable cell clustering based on gene expression.

• Sample Compatibility:

  • Single-Cell Nucleus RNA Sequencing: Can use frozen samples and is not limited by cell size, making it applicable to a broader range of samples.

• Data Analysis:

  • The data analysis for both methods is similar, focusing on:
    • Cell Clustering: Grouping cells based on gene expression profiles.
    • Cell Subpopulation Identification: Identifying specific subpopulations within the cell groups.
    • Marker Gene Identification: Determining genes that are indicative of specific cell types or states.
    • Gene Expression Analysis of Cell Clusters: Analyzing the gene expression patterns within different clusters.
    • Cell Trajectory Analysis: Studying the developmental trajectories and transitions between cell states.

• Suitable for Frozen Tissue Samples: Overcomes the limitation of needing fresh and timely samples required for single-cell transcriptome sequencing.

• Applicable to Tissues Difficult to Prepare as Single-Cell Suspensions: Effective for complex tissues such as brain, heart, and kidney.

• Ideal for Enzyme-Digested Samples with Bias: Provides a more accurate representation of cell type composition and proportions in samples where enzymatic digestion introduces bias.

• Appropriate for Samples with Large, Irregular Cells, or Low Cell Viability: Useful for samples with large cell sizes, irregular shapes, or low cell activity, such as neurons, cardiomyocytes, and skeletal muscle fibers.

• Suitable for Samples Where High-Quality Protoplasts Cannot Be Obtained: Enables single-cell level research by extracting nuclei when high-quality protoplasts are not achievable.