Boyden chamber assay

Pelz, G., Schettler, A., and Tschesche, H., Granulocyte chemotaxis measured in a Boyden chamber assay by quantification of neutrophil elastase. Eur. J. Clin. Chem. Clin. Biochem. J. 1993, 31 651 56. 

More functional studies that address the process of migration use the so-called Boyden chamber [48]. In this assay the migratory capacity of endothelial cells after activation with chemoattractants or pro-angiogenic stimuli is studied. Velocity of migration and the percentage of cells that are capable of migrating through an ECM-coated membrane into another compartment can be determined. 

Purification of chemokines from natural sources requires special strategies. First of all, the detection system should be useful to detect the required chemokine at low concentrations, i.e., it is important to choose an assay system that allows the detection of low amounts of the chemokine in a screening system. Screening systems often used are Boyden chamber chemotaxis assay... 

Multi-well modified Boyden chambers can be obtained from Neuro Probe Inc. (Gaithersburg, MD). Model AP48 has been widely used for endothelial cell chemotaxis assays. The apparatus consists of top and bottom acrylic plates, a silicon gasket and assembly screws. The bottom plate has 48 wells, each with 25 pL final volume. These correspond to holes on the top plate, and form the upper wells when the chamber is assembled. The filter (polycarbonate, 25 x 80 mm) is placed between the top and bottom plates, and a gasket is placed over the filter to create the seal. The apparatus can be purchased with a selection of accessories, such as curved forceps, filter clamps, and wipers. These are required to process the filters after use. Filters can also be obtained from Nucleopore Inc. (Pleasanton, CA) and Costar (Cambridge, MA). 

B42. Bignold, L. P., Ferrante, A., and Haynes, D. R. Studies of chemotactic, chemotactic movement-inhibiting and random movement-inhibiting effects of interleukin-1 alpha and beta, tumour necrosis factors alpha and beta and interferon gamma on human neutrophils in assays using sparse-pore polycarbonate (Nuclepore) membranes in the Boyden chamber. Int. Arch. Allergy Appl. Immunol. 91, 1-7 (1990). 

To determine the optimal concentration for collecting the invasive population, it is often necessary to perform dose response experiments and try a range of chemoattractant amounts. We have found that for EGF, the optimal concentration in the in vivo invasion assay is 25 nM while for the in vitro Boyden chamber chemotaxis it is 5 nM. The difference can be explained by different diffusion properties of the gradient emanating from a tip of a needle into live tissue vs. EGF in a buffer being directly accessible to migrating cells across pores in a filter. By the same token, the optimal... 

Chemokines promote the directional migration of cells expressing CKRs, which can be measured in chemotaxis assays using a Boyden chamber. Cells placed in the upper chamber, which is separated with a porous... 

Although the Boyden chamber and scratch assays have provided new insights into the mechanisms of cell migration, they are unable to achieve and maintain a linear gradient of signals. The visualization of cell migration cannot be achieved with Boyden chambers either. 

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