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Concept
Cell isolation
Résumé
Cell isolation is the process of separating individual living cells from a solid block of tissue or cell suspension. While some types of cell naturally exist in a separated form (for example blood cells), other cell types that are found in solid tissue require specific techniques to separate them into individual cells. This may be performed by using enzymes to digest the proteins that binds these cells together within the extracellular matrix. After the matrix proteins have been digested, cells remain loosely bound together but can be gently separated mechanically. Following isolation, experiments can be performed on these single isolated cells including patch clamp electrophysiology, calcium fluorescence imaging, and immunocytochemistry.
The techniques required to obtain isolated cells vary depending on the cell type required. Circulating cells such as blood cells or some tumour cells can be isolated by taking a blood sample. As blood samples contain a mixture of many different cell types, a method of separating out cells into different types must be used. The most commonly used method for this is flow cytometry, during which an automated analyser inspects a narrow stream of cells. In one version of this technique, a light is shone on the stream of cells, and the analyser detects the reflected light or fluorescence before using this information to rapidly manoeuvre the cells of interest into a collection chamber.
When dealing with solid tissues, obtaining tissue for cell isolation may be more challenging. Surplus human tissue can sometimes be obtained at the time of planned surgery, for example specimens of right atrial appendage are often excised and discarded during open heart surgery such as coronary artery bypass surgery. Other tissues such as samples of pancreas or bladder may be taken as a biopsy. Alternatively, tissue from animals is frequently obtained by sacrificing the animal.
After a tissue specimen has been obtained, it must be surrounded or perfused by a solution at an appropriate temperature containing the salts and nutrients required to keep the cells alive.
Source officielle
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