Page 54 - Live-cellanalysis handbook

Live-Cell Analysis Handbook — Third Edition

Kinetic Chemotaxis Assays

Powerful assays for quantification and visualization
of chemotactic cell migration and invasion

®
Introduction IncuCyte Chemotaxis Assay at a Glance
Chemotactic migration and invasion play an essential role in many The IncuCyte® ClearView chemotaxis cell migration assay is
normal and pathological processes including immune response, cell conducted in a 96-well migration plate consisting of an optically
differentiation and tumor metastasis. Methods to study factors clear membrane insert and reservoir that allows for direct
that regulate the directional migration of cells typically employ visualization of cell migration and morphological changes using
modifications of Boyden chamber assays. Boyden chamber assays phase contrast and/or fluorescent imaging. The plate is placed
consist of an upper and lower chamber separated by a porous in the IncuCyte® instrument, a fully automated live-cell analysis
filter, allowing for the diffusion of a chemotactic agent. Although system with integrated image analysis tools that is placed inside
these assays offer insight into the chemotactic response of cells, a standard tissue culture incubator. Integrated metrics precisely
they have inherent drawbacks including the inability to visualize quantify the chemotactic response using 1,000 to 5,000 cells
active cell migration and the requirement for a large number of per well, a significant advantage when using rare primary
cells, typically 50,000 to 100,000 cells per well. Most importantly, hematopoietic cells from the blood. This assay method is sensitive
perhaps, Boyden chamber assays “only afford the measurement to surface-integrin signaling, and sustains a linear gradient over
of cell migration at a single time point, and as the kinetics of cell several days. The result is 96 wells of reproducible kinetic data
migration vary markedly depending on the cell type studied and the supported by images and movies, requiring very few cells and
chemoattractant used, this limits the utility of this type of assay.” 1 minimal hands-on time.

In this chapter, we will review live-cell chemotatic cell migration
and invasion assays, which accurately measure and visualize cell
movement in response to a chemical gradient. This approach
allows for the assay to be completed in a physiologically relevant
environment, supports a stable gradient, and automatically
analyzes images, alleviating technically challenging and
cumbersome quantification steps in traditional approaches.

52