What Is Single Cell Analysis?
Recent advances in genome analysis technology have revealed that even seemingly homogeneous cell populations are actually heterogeneous when individual cells are isolated and analyzed. Single cell analysis provides insights into the unique characteristics of individual cells, leading to significant applications in fields like cancer research, immunology, and drug discovery. This approach allows for the exploration of cellular responses at an individual level, moving beyond the average values obtained from traditional population-based genetic analysis.
Method to Separate Cells for Single Cell Analysis
Single-cell separation can be done manually, but this method has a low throughput. For high-throughput processing, microfluidics is used to trap cells in chambers that can hold only one cell. However, the number of cells that can be trapped is limited by the plate area used to create the trap, usually capping at a few hundred cells. To process a larger number of cells, droplet-based cell separation systems have recently become the standard.
In droplet-based systems, fluorinated oil is used as the continuous phase, with hydrogel or lysis buffer as the dispersed phase. This setup enables the encapsulation of a single cell within each droplet, creating thousands of droplets per second. Hydrogel encapsulation is particularly useful for 3D culture, cellular response studies, and FACS, while encapsulation with specialized barcode beads is employed for mRNA capture and gene expression analysis.
Droplet-Based Principle for Single Cell Separation
The droplet-based method involves creating droplets at a junction point in a microfluidic channel. This is achieved by pumping a cell suspension, cell lysate, and fluorinated oil. The cell suspension and lysate are mixed just before encapsulation, ensuring that cells are lysed within the droplet. This method allows for precise and efficient single-cell encapsulation, facilitating detailed cellular analysis.