Blue cells on black background showing a wound healing scratch, with cells to the left and right.

Wound Healing Assay | Scratch Assay

Your automated and non-invasive wound healing assay

HoloMonitor wound healing assay icon

The label-free HoloMonitor® Wound Healing Scratch Assay enables cell-friendly, non-invasive cell migration analysis. It continuously images your samples, identifies every cell, and automatically analyzes the results. As a result, you receive accurate, kinetic gap closure data in real-time. In addition, combined with Culture-Inserts from ibidi®, you achieve superior reproducibility and consistency between your wound healing experiments.

Time-lapse video of a closing wound healing gap, captured with HoloMonitor.

Benefits

Why choose the HoloMonitor Wound Healing Scratch Assay?

Accurate

Get exact cell segmentation with digital holography to measure what you actually want to study.

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Non-invasive

Avoid known interferences from cellular markers and achieve biologically relevant results.

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User-friendly

Save time with the automated and guided App Suite software, designed by and for cell biologists.

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Reproducible

Combine this assay with the ibidi Culture-Inserts for an easy and consistent experimental setup.

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Assay Output

What results do you get with the HoloMonitor Wound Healing Scratch Assay?

The HoloMonitor Wound Healing Assay automatically determines your gap closure kinetics in real-time. You can follow the process in a time-lapse video and the software will provide you with the relevant data for each time point.

  • Kinetic gap closure data over time
    • Gap width [µm]
    • Cell-covered / cell-free area [µm², %]
    • Cell front velocity [µm/h]
  • Images and time-lapse videos of the gap closure process
  • Combine with the Single Cell Tracking Assay and Cell Morphology Assay for deeper insights
A line chart from a wound healing assay showing a decreasing gap width with time. The slope equals the cell front velocity (23.4 µm/h)..
Wound healing assay gap closure graph with linear regression (cell front velocity).
A line chart from a wound healing assayshowing an increased cell-covered area over time.
Wound healing assay cell-covered area graph.
Blue cells on white background showing a closing cell gap of a wound healing assay
3D image of the wound healing gap. The cells are not stained in any way. In fact, the colors are chosen in the software.

Label-free time-lapse video showing non-treated human breast cancer cells (JIMT-1) closing a wound healing gap.

Image showing black and white cells to the left, and the same cells segmented to the right.

Accurate measurements.

Exact cell segmentation measures what you actually want to study

Compared to traditional phase contrast microscopy, the quantitative phase imaging technology used by HoloMonitor offers superior cell segmentation and identification. As a result, you get a distinct separation between cells and the background. This enables the HoloMonitor software to provide accurate data, in 3D, on every single cell. Therefore, using HoloMonitor, you can be sure to measure what you actually want to study.

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On-demand Webinar

Watch our Wound healing assay webinar together with ibidi!

This webinar showcases how you how to run wound healing assays that get you biologically relevant and reproducible data using the HoloMonitor Wound Healing Assay protocol.

Key Applications

What can you study with the HoloMonitor Wound Healing Assay?

Wound healing (scratch) studies

Transwell studies

Co-culture studies

Cell invasion studies

Cell-friendly wound healing analysis.

Study your cells non-invasively right inside your standard incubator

The non-invasive quantitative phase imaging technology used by HoloMonitor does not affect your cells in any way. In fact, you image your wound healing experiments without any labels or stains. Moreover, the low-energy laser keeps phototoxicity at a minimum. In addition, everything takes place in your standard incubator. As a result, by eliminating elements known to influence cellular behavior, you are sure to get biologically relevant results.

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Wound Healing Assay Protocol

How do you set up this in vitro wound healing assay?

1. Sample preparation

  • Prepare your wound healing cell cultures as you normally would. With ibidi’s Culture-Inserts, you will get a defined gap suitable for HoloMonitor.
  • Use HoloLids for optimal image quality and data acquisition.
  • Click your cell culture vessel onto the HoloMonitor motorized stage and you are ready to go!

2. Assay setup

  • Select the Wound Healing Assay in the software and follow the guided workflow. Next, name your experiment and define treatments.
  • Add the positions you want to image. Also, choose how often and for how long you want to image your cells.
  • Start imaging and walk away!

3. Explore results

  • Receive kinetic gap closure data and time-lapse videos in real-time. Analyze the results with other live cell assays for deeper insights into your cells.
  • Export the raw data to Excel for further analysis and customized data presentation.
  • Re-use the untouched cells for other experiments!

Reproducible setup.

Consistent setup with non-biased data collection and analysis

The HoloMonitor Wound Healing Assay gets you relevant results from controlled wound healing conditions. Firstly, the ibidi Culture-Inserts allow for easy and consistent gap creation. Secondly, HoloMonitor software offers a streamlined software setup. In addition, the whole assay takes place in the stable environment of your standard incubator. Moreover, you can use HoloLids™ to achieve optimal image quality.

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Smiling cell biologist closing an incubator in the lab

Easy-to-use, yet powerful, software.

Save time in your lab with automation

The HoloMonitor App Suite cell imaging software is designed by and for cell biologists. In fact, the user-friendly workflow guides you through the whole wound healing assay, from setup to data presentation. Also, with pre-defined templates for the ibidi Culture-Inserts, you can set up in no time. In addition, the system automatically performs image acquisition, analysis, and data presentation. So you can follow your experiment in real-time.

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Get more data from your samples

A major benefit with HoloMonitor is that you can get more data from your samples compared to traditional methods. Not only does it leave your cells unaffected. But it also allows you to reanalyze your already collected image sequences in new ways.

Pipetting with Hololid in 6-well microplate

Re-use your cells after the imaging is done

Above all, HoloMonitor technology images your cells without affecting them in any way. As a result, they are completely untouched when the experiment is over. This means that you can use the very same cells for subsequent experiments. Therefore, you save both time, money, and cells.

Scatter plot of L929 etoposid treated cells

Re-analyze your data in new ways

Moreover, data sets recorded with the Wound Healing Assay can be used for other App Suite analyses as well. For example, if you do a gap closure experiment and realize that you need proliferation or morphology data, you can simply re-analyze the old imaging experiment with another HoloMonitor Assay. In other words, there is no need to redo the experiment. And you will get data on the exact same cells as your original study.

Deeper insights from your wound healing experiment

Certainly, the HoloMonitor Wound Healing Assay can be combined with other HoloMonitor Assays. Therefore, you can get an even better understanding of your experiments.

Image showing a closing wound healing gap, where cells at the fronts are tracked over time.

With the Single Cell Tracking Assay, you can track every single cell in your image sequence and get detailed data on cell movement and cell division. This lets you detect unique cell behavior on a single-cell level and reveals not only when, but also how, the gap was closed.

Screenshot showing a cell feature plot of optical volume in HoloMonitor App Suite

Moreover, you can study over 30 different cellular features with the Cell Morphology Assay. For example, you can explore cell volume or thickness, down to the single-cell level. In addition, the results can be visualized in a dot plot. And this may reveal sub-populations of cells that behave differently in your gap closure experiment.

The advantage of this compared to traditional gap closure analyses is that it is a high throughput system, uses standard culture conditions with an automated image capture, segmentation and tracking of cells and combines functional aspects of cellular behavior with morphological properties on cell level.

Sofia Mogren et al.
Mast cell tryptase enhances wound healing by promoting migration in human bronchial epithelial cells, Cell Adhesion & Migration (2021)

Browse our publication library for more references and see how our worldwide HoloMonitor users run their wound healing analysis.

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