AccuWound 96

Cell migration/wound healing kinetics in a highly reproducible 96-well format.

Is Your Wound Healing Assay Due for an Upgrade?

Cell migration and invasion are critical to many facets of biology, including such diverse processes as wound healing, angiogenesis, embryonic development, and the intravasation and extravasation of cancer cells during metastasis. The traditional scratch assay/wound healing assay, a staple for cell migration studies, suffers from tedious and low throughput setup, requires repetitive visual evaluation of cells, and provides subjective data with poor reproducibility. Combining the xCELLigence® Real-Time Cell Analysis instruments with the new AccuWound® 96 scratch tool overcomes these deficiencies and has completely revolutionized the wound healing assay:

  • Generate 96 Identical Scratch Wounds in Seconds: With a simple turn of a knob, pins on the AccuWound 96 tool produce uniform scratch wounds in every well of a microtiter plate.
  • Astonishingly Simple Data Acquisition and Kinetic Analysis: Once scratch wounds have been generated, the xCELLigence instrument monitors cell migration/wound healing in real-time:
    • Read an entire 96-well plate in 15 seconds
    • Run up to 6 plates simultaneously yet independently
    • Track cell migration/wound healing continuously, providing a true kinetic analysis and eliminating the guesswork that is associated with endpoint data
  • Exceptional Reproducibility: The precision of the AccuWound 96 scratching tool, combined with the objective and continuous monitoring of the xCELLigence instrument, yield data of unparalleled quality. Coefficients of variation are typically 4% (vs. 20% for traditional scratch assays).
Example Data

Though not necessary for the xCELLigence wound healing assay, cells were stained and visualized (above photos A-E) in order to demonstrate the correlation between biosensor coverage and the magnitude of the Cell Index signal (data plot to the right). Key features of the Cell Index plot, including the specific time points corresponding to the above photos A-D, are denoted.

 

 

 

 

 

The xCELLigence® Real-Time Cell Analysis (RTCA) instruments utilize patented microtiter plates (E-Plates®) which contain gold biosensors embedded in the bottom of each well. These biosensors continuously and non-invasively monitor the total surface area that is covered by cells, enabling the kinetics of wound closure to be determined. The simple workflow is as follows:

Step 1 – Cell Seeding and Growth: Adherent cells are first seeded in E-Plate wells, and the E-Plate is placed inside the xCELLigence instrument. The xCELLigence biosensor signal, also known as “Cell Index”, increases as cells attach and proliferate, and then plateaus as cells approach 100% confluence (above photo A and data plot).

Step 2 – Scratch Wound Production: The E-Plate is removed from the xCELLigence instrument and placed inside the AccuWound 96 scratch tool (shown on top of this page). Turning a knob slides a spring-loaded Teflon™ pin across the bottom of each well in the E-Plate. This removes cells from the biosensors (above photo B), which xCELLigence subsequently detects as a drop in the Cell Index signal (above data plot).

Step 3 – Monitoring Cell Migration/Wound Healing: As cells migrate to close the wound, they progressively re-coat the gold biosensors (above photos C-E), causing the Cell Index signal to increase (above data plot). The kinetics of this process are easily compared for different conditions (± drug, gene knockdown, etc.)

 

xCELLigence RTCA systems compatible with the AccuWound 96 Scratch Tool:

Single PlateMulti Plate
RTCA SPRTCA MP
1×96 wells6×96 wells

Real Time Chemotactic Cell Migration Assays Can Also be Run with ACEA’s Electronic Boyden Chamber. Learn more here.