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The migration of cultured cells attached to a surface or in 3D is commonly studied using microscopy. [7] [8] [5] As cell movement is very slow, a few μm/minute, time-lapse microscopy videos are recorded of the migrating cells to speed up the movement.
In general, the most important requisite is to calibrate the incubation time of the assay both to the model cell and the ligand to be evaluated. Too short incubation time results in no cells in the sample, while too long time perturbs the concentration gradients and measures more chemokinetic than chemotactic responses.
Migration of cells is free toward and away on the axis of the concentration gradient; Detected responses are the results of active migration of cells; Despite the fact that an ideal chemotaxis assay is still not available, there are several protocols and pieces of equipment that offer good correspondence with the conditions described above. The ...
Consequently, in cells containing the plasmid with an insert, no functional β-galactosidase may be formed. The presence of an active β-galactosidase can be detected by X-gal , a colourless analog of lactose that may be cleaved by β-galactosidase to form 5-bromo-4-chloro-indoxyl, which then spontaneously dimerizes and oxidizes to form a ...
The scratch assay is a great tool to study cell migration since this mechanism is involved in many different physiological aspects. [7] Cell migration plays a huge role in re-epithelialization of the skin and so the study of cell migration can provide advancements in understanding non-healing wounds. [7]
The most common technique to add inserts to desired sequences is the use of homologous recombination. [5] This technique has a specific requirement where the insert can only be added after it has been introduced to the nucleus of the cell, which can be added to the genome mostly during the G2 and S phases in the cell cycle. [6]
The PiggyBac (PB) transposon system employs a genetically engineered transposase enzyme to insert a gene into a cell's genome. It is built upon the natural PiggyBac (PB) transposable element (transposon), enabling the back and forth movement of genes between chromosomes and genetic vectors such as plasmids through a "cut and paste" mechanism.
Schematic of cell adhesion. Cell adhesion is the process by which cells interact and attach to neighbouring cells through specialised molecules of the cell surface. This process can occur either through direct contact between cell surfaces such as cell junctions or indirect interaction, where cells attach to surrounding extracellular matrix, a gel-like structure containing molecules released ...
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