IncuCyte® Chemotaxis Cell Migration Assay Frequently Asked Questions

Assay compatibility

Q: What is the difference between the IncuCyte® Scratch Wound and Chemotaxis Cell Migration Assays?

A: The IncuCyte® Scratch Wound Assay measures the movement of cells into a cell-free zone in the absence of a chemotactic gradient. You can use the WoundMaker® to create a precise wound in each well and monitor wound closure as cells move either across a substrate (migration) or through a 3D gel matrix (invasion).

The IncuCyte® Chemotaxis Assay measures directed cell migration in response to a chemotactic gradient. You can use the IncuCyte® Live-Cell Analysis System to investigate treatment effects on chemotactic profile. The system is suitable for adherent and non-adherent cells.

Q: Can I use my own Boyden chamber plate with the IncuCyte® Live-Cell Analysis System?

A: No. The optical quality of the membrane surfaces in existing Boyden chambers is not amenable to IncuCyte® imaging or image processing algorithms.

Q: Are the ClearView inserts compatible with any plate?

A: No. The ClearView cell migration insert has been specifically designed to work with the ClearView reservoir plate.

Q: What IncuCyte® software version must I use?

A: Version 2015A Rev 1 or later. The IncuCyte® Chemotaxis Analysis Software Module is not compatible with prior versions of software. If you wish to receive a software upgrade, please contact your point-of-sale contact.

Q: Can I perform the IncuCyte® Chemotaxis Cell Migration Assay using an IncuCyte® FLR?

A: No. The IncuCyte® FLR instrument does not have the ability to acquire or process the images quickly enough.

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Experimental setup

Q: Can I measure migration of non-adherent cell types using the ClearView cell migration plate?

A: Yes. The ClearView Cell Migration plate has been extensively validated with non-adherent cell types. To ensure uniform cell settling within each well after seeding. We recomend allowing the plate to sit at room temperature for 45-60 minutes after plating. The Chemotaxis (Top Only) “Analysis Job Type” is recommended for quantifying non-adherent cell migration.

Q: How do I optimize the IncuCyte® Chemotaxis Assay for my cell type of interest?

A: Key variables to optimize include membrane coatings, assay medium and cell density. Recommended experimental conditions for a range of cell types, including cancer cells, vascular cells, and leukocytes, can be found in the assay protocol.

Q: Am I able to coat both sides of the membrane?

A: Yes. Both the top and the bottom membrane surfaces can be coated independently or together. Recommended volumes are 20 μL for the top side of the membrane and 150 μL for the bottom side of the membrane.
Learn how to coat

Q: Can I measure chemotactic cell migration using fluorescently labelled cells?

A: Yes. We recommend using a nuclear restricted fluorophore (as opposed to cytoplasmic) to ensure neighboring cells can be differentiated and counted as individual objects following image analysis. Fluorophore excitation and emission spectra must be compatible with IncuCyte® fluorescent channels.

Q: Could I use the IncuCyte® Chemotaxis Assay to study chemotaxis in co-cultures?

A: Yes. By labelling different cell types with fluorescent probes and using the phase, green and red data acquisition channels of IncuCyte® system it is possible to quantify chemotaxis of more than one cell type in co-culture.

Q: Is it possible to remove medium from the wells to perform biochemical analysis?

A: Yes. The IncuCyte® Live-Cell Analysis platform does not disturb your cells during the assay. Once the experiment is completed, simply remove the medium from the wells to perform additional analyses.

Q: Can I make parallel measurements of cell health using a fluorescent probe?

A: Yes. Multiplexed measurements of cell health can be made using any unused imaging channel. The IncuCyte® Live-Cell Analysis System supports HD phase-contrast, green fluorescence and red fluorescence automated imaging modes.

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