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Boyden Chamber Technique

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Boyden chamber technique protocol.
Cell migration may be evaluated through several different methods including scratch assays, cell-exclusion zone assays, microfluidic based assays, and Boyden Chamber assays. The most widely accepted cell migration technique is the Boyden Chamber assay. The classic Boyden Chamber system uses a hollow plastic chamber, sealed at one end with a porous membrane. This chamber is suspended over a larger well which may contain medium and/or chemoattractants. Cells are placed inside the Chamber and allowed to migrate through the pores, to the other side of the membrane. Migratory cells are then stained and counted.

Cell invasion into a pore.
In a standard Boyden assay, the pore diameter of the membrane is typically 3 to 12 μm, and is selected to suit the subject cells. Smaller pore size results in a greater challenge for the migrating cell. Most cells range in size from 30-50 μm and can migrate efficiently through 3-12 μm pores, whereas, lymphocytes (10 μm) can migrate through pores as small as 0.3 μm.

Pore Size Matters!
How to select the appropriate pore size for your cells:
  • 3 μm pore size is appropriate for leukocyte or lymphocyte migration.
  • 5 µm pore size is appropriate for a subset of fibroblast cells or cancer cells such as NIH-3T3 and MDA-MAB 231 cells. Also suitable for monocytes and macrophages.
  • 8 μm pore size is appropriate for most cell types. This pore size supports optimal migration for most epithelial and fibroblast cells. Note - the 8 μm pore size is not appropriate for lymphocyte migration experiments.
Click here to match your cell type to its suitable assay pore size.