Several methods are employed for cell isolation, each suited for different types of cells and tissues. Some common techniques include:
Mechanical Disruption This method involves physically breaking down the tissue to release individual cells. Techniques such as mincing with scalpels, or using blenders and sieves, are commonly used. Mechanical disruption is simple but may cause damage to delicate cells.
Enzymatic Digestion Enzymes such as collagenase, trypsin, and dispase are used to degrade the extracellular matrix, facilitating the release of individual cells. This method is gentler on cells compared to mechanical disruption but requires optimization to avoid over-digestion.
Density Gradient Centrifugation This technique separates cells based on their density. A sample is layered over a density gradient medium and centrifuged. Cells migrate to the region of the gradient that matches their density. Common media include Ficoll and Percoll.
Fluorescence-Activated Cell Sorting (FACS) FACS uses fluorescently labeled antibodies to target specific cell surface markers. Cells pass through a laser beam, and those with the target fluorescence are separated. FACS provides high purity and specificity but requires specialized equipment.
Magnetic-Activated Cell Sorting (MACS) MACS utilizes magnetic beads conjugated with antibodies to target specific cell types. The cell-bead complexes are separated using a magnetic field. This method is relatively quick and simple but may have lower purity compared to FACS.
