Page 49 - Live-cellanalysis handbook

Kinetic Scratch Wound Assays

Real-time assays to assess migration and invasion potential

Introduction IncuCyte® Scratch Wound Assay at a Glance

Cell migration is a multistep process that is a fundamental The IncuCyte® scratch wound migration and invasion assay can
component of many biological and pathological processes such be used to quantitatively assess both cellular migration and cell
as embryonic development, tissue re-organization, angiogenesis, invasion potential in the presence of experimental agents. The
immune cell trafficking, chronic inflammation and tumor 96-well IncuCyte® WoundMaker creates cell-free zones in cell
metastasis. Cell migration is initiated by a stimulus that activates monolayers cultured on IncuCyte® ImageLock 96-well plates
a set of signaling pathways leading to cellular polarization and with the use of precision engineered pins, without disrupting
a rapid reorganization of actin filaments and microtubules. Cells the underlying biomatrix or damaging the tissue culture treated
advance by protruding their membrane at their leading cell border, plastic. After wounding, the scratch wound assay is initiated by
which is followed by dynamic substrate adhesion via integrin simply adding media to the cells for the migration assay or by
adherence to the substrate. Membrane retraction at the lagging overlaying the cells with an optimized concentration of a biomatrix
cell edge finishes the cycle, which is then repeated in rapid material for the invasion assay. IncuCyte® scratch wound analysis
succession. The summation of this process results in cell migration. software enables real time, automated measurement of label-free
or dual fluorescence of cell migration and invasion.
In this chapter, we will review real-time visualization and analysis
of cell movement that is driven by changes in cell density. Unlike • Visualize cell migration and invasion, and assess morphological
traditional assays, which require repeatedly removing cells from changes in real time with the scratch wound method.
an incubator to perform manual image acquisition and have • Assays are flexible, quantitative and easily reproducible with the
variability in scratch width and uniformity, this live-cell assay IncuCyte WoundMaker tool.
approach provides continuous analysis and eliminates assay
variability with the use of the WoundMaker tool to create precise • Monitor and quantify cell movements across a substrate
scratches in the cell monolayer. (migration) or through a gel matrix (invasion) in the same plate,
up to six 96-well plates at once.
• Validated with a wide range of adherent primary, immortalized
and tumor cell types.

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