Automated wound healing and migration assay monitoring — 24 wells in parallel, continuous kinetic data, real-time gap closure analysis. No manual imaging. No missed time points.
The wound healing, migration assay or scratch assay is a standard method to analyze cell migration in vitro. The dynamics of cell migration into a cell-free area are monitored and quantified to analyze migration characteristics and calculate gap closure time.
With conventional manual microscopy it is not possible to capture every dynamic change at the cell level. zenCELL owl records images every 5 minutes — capturing every cell movement in detail, even during long-term analyses over days.
Observe and control every time point of the wound healing process retrospectively. Calculate gap closure speed by analysing the increase of confluence in the gap area. Determine t½ gap closure time automatically.
Simultaneous analysis of cell morphology and confluence in up to 24 wells under identical incubator conditions. Optimises comparability of results and reproducibility of data across conditions, drugs and cell lines.
A migration assay of L929 fibroblasts was performed over 24 hours. The gap was inserted in a confluent monolayer at time zero. Within 24 hours, continuous cell migration results in complete closure of the gap — captured automatically at every time point.
L929 mouse fibroblasts — scratch assay timelapse over 24 hours. Gap inserted at 0h by pipette tip. Complete closure visible at 24h. Digital phase-contrast imaging. Scalebar: 200 µm.
Relative gap area as a function of time — t½ gap closure determined automatically from confluence data
Cell coverage data from zenCELL owl is used to calculate the Relative Gap Area at every time point. Plotted as a function of time, the half gap closure time (t½ Gap Closure) is determined directly from the confluency curve.
Application Note — Scratch Assay
Full protocol, gap closure analysis method and data for scratch assays with zenCELL owl.
From cell seeding to final data export — the complete automated scratch assay workflow with zenCELL owl. Every step documented, every time point captured.
Seed cells in 24-well plate and grow to full confluency. Use coated plates for optimal adhesion and consistent monolayer formation.
Use a pipette tip for standard assays — or the zenCELL Scratch Maker (light-based, ECM-preserved) for physiological migration studies. Full 24-well plate in 60 seconds.
Wash away detached cells. Add drug treatment, inhibitor, or growth factor at defined concentrations across 24 wells.
Transfer plate to zenCELL owl inside the incubator. Define imaging interval (5–30 min). Continuous monitoring starts automatically — no further manual steps needed.
Software calculates Relative Gap Area at every time point automatically. Growth curves generated per well. t½ gap closure determined from confluence data — no manual measurement.
Export CSV data and timelapse videos. Compare gap closure speeds across all 24 conditions. Full retrospective analysis of any time point — publication-ready data.
Not all scratches are equal. The method used to create the gap fundamentally affects the biology of the migration assay.
Full method comparison: Migration Assay Methods compared →
The example above uses a manual pipette tip scratch. For publication-quality, physiological migration data, the zenCELL owl Scratch Maker uses light-based technology — preserving cells and ECM completely.
A manual pipette tip scratch destroys the cell layer and ECM. The Scratch Maker uses photonic technology to create a precise, reproducible gap without any mechanical contact. 24 identical scratches in 60 seconds.
Watch zenCELL owl track gap closure in real cells inside a real incubator. On request via MS Teams. No obligation.
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