Detachment of cells refers to the process of releasing adherent cells from the surface of a culture vessel, such as a flask or dish, in order to subculture, harvest, or perform downstream experimental procedures. Detachment can be achieved through various methods, including enzymatic, mechanical, or chemical approaches, depending on the cell type, culture conditions, and experimental requirements. Here are some common methods for detaching cells:

1. Enzymatic methods: These methods involve using enzymes, such as trypsin, collagenase, dispase, or papain, to cleave proteins responsible for cell adhesion, like integrins and cadherins. One widely used enzyme is trypsin, often combined with EDTA, which chelates divalent metal ions, further weakening cell-substrate interactions. Enzymatic detachment requires careful monitoring of the enzyme concentration and incubation time to avoid damage to the cells.
2. Mechanical methods: These methods involve physically disrupting the cell-substrate interactions using tools such as cell scrapers, rubber policemen, or pipettes. Mechanical detachment can be less harmful to the cell surface compared to enzymatic methods, but it may not provide a uniform single-cell suspension and can be more labor-intensive.
3. Chemical methods: These involve using chemicals like ethylenediaminetetraacetic acid (EDTA) or ethylene glycol tetraacetic acid (EGTA) to chelate divalent cations, such as calcium and magnesium, which are essential for maintaining cell-substrate interactions. These chelating agents can be used alone or in combination with other detachment methods. Chemical detachment is generally more gentle on cells than enzymatic methods but may not be as effective for strongly adherent cells.
4. Temperature-based methods: Some cells can be detached by simply lowering the temperature of the culture, for example, by incubating the cells on ice. This method is less commonly used but can be suitable for some cell types, particularly those that are sensitive to enzymatic treatments.

The choice of detachment method depends on the specific cell type, culture conditions, and experimental requirements. It is important to select an appropriate method that minimizes damage to the cells and maintains their viability for downstream applications.