Wound Healing With Single-cell Tracking

The HoloMonitor® Wound Healing Scratch Assay analyze in vitro cell migration. Cell migration speed is automatically determined in a straightforward way, directly in your incubator.

The in vitro wound healing assay, also known as scratch assay, is widely used to model in vivo cell migration. The method is based on quantifying the rate at which cells repopulate an artificial gap or scratch created in a confluent monolayer of cells.

Single-cell Tracking

The HoloMonitor incubator microscope provides an automated label-free wound healing assay, measuring scratch closure rate and cell front velocity. Additionally, cell tracking may be applied on selected cell front cells for detailed individual cell movement and morphology analysis.

After recording the wound healing time-lapse image sequence, individual cells at the cell front may be tracked to determine their migration direction with the HoloMonitor Single-cell Tracking Assay. Detailed, non-biased data on migration into the wound area is then easily achieved. When preferred, further analyses can be undertaken for details on cell morphology changes, cell proliferation and cell division using the HoloMonitor Live Cell Assays.

Wound Healing Assay Image and Graph

The HoloMonitor Wound Healing Assay provides kinetic gap closure analysis showing how gap width and cell covered area change over time.

Videos showing gap closure over time as the cells fill the gap. Untreated cells are used and the gap is created using the ibidi® Culture-Insert 2 Well.

HoloMonitor Wound Healing Assay with single-cell tracking

Tracked cell front cells, overlayed tracks (left), migration plot (right).

HoloMonitor® Wound Healing Assay — Get the Whole Picture

The assay is based on a reproducible method to create the gap, coupled with graphics and automated data analysis to provide gap closure rate. As with all HoloMonitor applications, it is cell-friendly and based on label-free live cell imaging.

  • Cell front velocity
  • Time-lapse videos showing how cells migrate into the gap
  • Automatic and kinetic calculation of gap closure over time
  • Single-cell tracking

Key References

  • Moving into a New Dimension: Tracking Migrating Cells with Digital Holographic Cytometry in 3D
    A. Görloff Wingren
    Cytometry Part A (2018)

    Commentary article discussing the fundamental role of cell movement studies in cancer research. Advantages of the HoloMonitor Cell Tracking and Wound Healing Applications over the transwell migration and invasion assays are highlighted, and includes the possibility to use the cells in experiments for other purposes after completing the imaging. In addition to single cell tracking, the HoloMonitor technology also benefits from the fact that morphology analysis can be performed of each cell. The author concludes that this indeed opens up for almost unlimited possibilities to perform cell morphology analysis using this methodology, since each image is very rich in cellular information.

    Read the article
  • Evaluation of Holographic Imaging Cytometer HoloMonitor M4 Motility Applications
    Y. Zhang and R. L. Judson
    Cytometry Part A (2018)

    The HoloMonitor software modules for cell tracking and wound healing analysis were evaluated and compared to the more conventional methods transwell migration and transwell invasion. Both HoloMonitor modules were found to be well-correlated with established standards, yielded reproducible results, and at the same time offered distinct advantages. The wound healing assay was the most tractable and automated method with good reproducibility, while the cell tracking module enabled identification of hypermobile subpopulations.

    Read the article