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In vitro Cell Migration and Invasion Assays
Published on: June 1, 2014
Modifications to the Transwell Migration/Invasion Assay Method That Eases Assay Performance and Improves the Accuracy
Heidi Marie Stoellinger1, Arshak R Alexanian1
1Cell Reprogramming & Therapeutics LLC, Wauwatosa, Wisconsin, USA.
Assay and Drug Development Technologies
|February 23, 2022
Summary
Improving cell migration assays is crucial for understanding cancer metastasis and immune responses. This study refines the transwell assay, enhancing accuracy by counting cells on top of the membrane using DAPI staining and image overlays.
Area of Science:
- Cell Biology
- Biomedical Research
Background:
- Cell migration is vital for development, immunity, and diseases like cancer metastasis.
- In vitro assays are essential for studying live cell behavior and extrapolating to in vivo conditions.
Purpose of the Study:
- To discuss existing transwell migration/invasion assay methods.
- To identify problems and propose modifications for improved simplicity and accuracy.
Main Methods:
- Modification of the standard transwell migration assay.
- Utilizing transparent transwell inserts.
- Employing 4',6-diamidino-2-phenylindole (DAPI) staining for cell visualization.
- Overlaying bright light and DAPI images for enhanced analysis.
Main Results:
- Accurate evaluation of migrated/invaded cells requires counting cells on the membrane's top surface.
- Transparent inserts with DAPI-stained cells simplify analysis of cells on membranes.
- Overlaying bright light and DAPI images further improves cell count accuracy.
Conclusions:
- The proposed modifications simplify the transwell assay procedure.
- These enhancements significantly improve the accuracy of the transwell migration assay method.
- The refined method aids in the study of cell migration in various biomedical research fields.
