Q: What cell lines are compatible with this assay?
A: Our Wound Healing Assay is compatible with any adherent cell line.
Q: Can the inserts be used with a different plate?
A: We don’t recommend using a different plate with the inserts. The wound inserts are custom made with a plastic precision injection mold to create the 0.9mm wound gap specifically for the kit plate. It is critical that the insert fits properly and stays in contact with the plate at all times during the formation of the monolayer to get a good wound gap. We have screened other plates and found that most do not provide a clean wound gap.
Q: Are the inserts reusable?
A: The inserts are not reusable and autoclaving them can change their dimensions, which can also affect the quality of the wound gap.
Q: How can I get a clean gap?
A: To create a clean wound gap, the insert and the 24-well plate must stay in contact during the monolayer formation. There are a few critical steps to using this assay:
1. It is important not to seed too many cells. Before performing the wound healing assay, a cell dose curve can be performed in a regular 24-well plate to find out how many cells are needed to form a monolayer within a certain time (such as overnight incubation).
2. After adding cells, close the plate lid and gently press the lid once. After that, you don’t want to touch it or do anything until the monolayer formation is complete. If possible, do not move the plate until the inserts are ready to be removed.
3. Any media change should be done after the inserts have been removed.
Q: Is an inverted microscope necessary for this assay?
A: Yes, this assay requires the use of an inverted microscope, either a light microscope or a fluorescence microscope with a DAPI filter.
Q: Can the results from this kit be quantified?
A: The percent closure or migration rate can be determined using either a light or fluorescent microscope. There are detailed instructions in the product manual on how to measure percent closure (pg 6-7). Briefly, you will measure a defined area of the wound and monitor wound closure with either a light microscope or imaging software. You have the option to visualize cells with a phase contrast microscope, DAPI fluorescent labeling, or cell staining.
Q: What image analysis software do you recommend?
A: We recommend NIH Image, Image Pro Plus, or CellProfiler.