Featured Technical Note
|Image analysis of the u-Migration Assay
A detailed protocol for data collection and analysis of the real-time migration of single cells.
The Millicell® µ-migration assay kit is a powerful, slide-based platform that measures the effects of chemoattractants on the migration of adherent single cells through real-time imaging. A stable, linear concentration gradient is established for more than 48 hours and parameters such as single cell directionality and velocity can now be measured. This innovative platform results in high content data and analysis not achievable with Boyden chambers or wound healing assays.
Traditional multiwell (Boyden chamber) migration assays can have limitations, such as: (1) gradients are not linear, well-established, or controlled, (2) quantification of migratory cells is based on an endpoint measurement rather than a dynamic determination, and (3) imaging of the cells while they are migrating is currently not possible.
The Millicell® µ-Migration Assay Kit overcomes these limitations. Its microfluidic, low-volume technology promotes a stable, diffusion-generated concentration gradient that is consistently linear and lasts for more than 48 hours. Designed for video microscopy assays, the Millicell® µ-Migration Slide is made from a plastic with high optical qualities similar to those of glass. At specific time intervals, images of the observation area can be acquired, allowing real-time monitoring and quantitative measurements of cell migration.
Analyze three chemoattractants in parallel for increased throughput and flexibility. There's no need to spend time or money on optimization as the kit comes ready-to-use, with no assembly needed.
Enhance your studies of metastasis, development, angiogenesis, and more with the Millicell® μ-Migration Assay Kit by distinguishing chemotaxis from random movement. Enhanced optical imaging for slow and fast-migrating cells and multiparametric analysis results in greater mechanistic insight. HT-1080, HUVEC, MDA-MB-231, and NIH-3T3 cell lines have been shown to work effectively with the Millicell® µ-migration kit.
Move your research and your cells forward A stable, linear concentration gradient
The concentration profile remains linear over the 1 mm observation window after 1 hour and is linear and stable for over 48 hours. Image examples of flourescence measurements Relative intensity across a 1 mm slit (raw and smoothed data)
**The calculated maximum concentration is only 70% of the applied concentration. The final maximum concentration is only 33% of the applied concentration. Compare this with a Boyden chamber where very steep gradients may form along a single axis perpendicular to the surface of the membrane.