AffiEXTRACT® Single Nuclei Isolation Kit for Neuronal Tissues & Cells

The "AffiEXTRACT® Single Nuclei Isolation Kit for Neuronal Tissues & Cells" is designed to extract individual nuclei from a brain tumor sample (or any neuronal tissue) for downstream analysis, like single-nucleus RNA sequencing (snRNA-seq). 

Step-by-Step Process:

1. Sample Preparation:

A small piece of the brain tumor tissue is collected. Since brain tissue, especially in tumors, can be dense and hard to break down into individual cells, isolating nuclei is a better approach.

2. Tissue Dissociation:

The tissue is gently broken into smaller pieces. This is important for freeing the nuclei without causing damage. Unlike whole-cell isolation, this process focuses on getting intact nuclei, not entire cells.

3. Nuclei Isolation:

The kit uses special reagents to release the nuclei from the cells. In tough neuronal tissues, these reagents break the cell membranes while leaving the nuclear membranes intact, ensuring the nuclei are ready for analysis.

4. Nuclei Purification:

The isolated nuclei are purified to remove unwanted cell debris, cytoplasmic material, and other impurities. Depending on the kit's protocol, this might involve centrifugation or filtration steps to ensure high-quality nuclei.

5. Nuclei Counting & Quality Check:

The next step is to check the number and quality of the nuclei. This can be done using a microscope or an automated counting device to ensure they are intact and ready for sequencing.

6. Downstream Application (Single-Nucleus Sequencing):

Once isolated, these nuclei are ideal for single-nucleus RNA sequencing (snRNA-seq). This is particularly useful when whole-cell isolation is difficult, like in dense brain tumor tissues.

Using snRNA-seq, researchers can profile gene expression in individual nuclei, helping identify specific cell types, subpopulations, or gene expression changes that may drive tumor growth.

Why Use This Kit for Brain Tumor Samples?

• Works with Dense Tissue: Brain tumors are difficult to break into whole cells, making nuclei isolation a more effective approach.
• Preserves Nuclear Integrity: The kit ensures that nuclei stay intact during isolation, providing high-quality samples for sequencing.
• Helps Study Tumor Heterogeneity: Tumors have many different cell types. By isolating individual nuclei, researchers can explore the diversity of cells in the tumor, which might reveal insights into tumor growth, resistance, or metastasis.

Applications in Brain Tumor Research:

Using isolated nuclei, researchers can:

• Analyze how different cells in the tumor express genes.
• Identify mutations or pathways that drive tumor growth.
• Study the tumor environment by examining the surrounding neuronal cells.

In summary, the kit provides an efficient way to isolate high-quality nuclei from brain tumor tissue, allowing for detailed genetic analysis at the single-cell level. This can give researchers a deeper understanding of tumor biology and potential therapeutic targets.