In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro
Abstract
The in vitro scratch assay is an easy, low-cost and well-developed method to measure cell migration in vitro. The basic steps involve creating a "scratch" in a cell monolayer, capturing the images at the beginning and at regular intervals during cell migration to close the scratch, and comparing the images to quantify the migration rate of the cells. Compared to other methods, the in vitro scratch assay is particularly suitable for studies on the effects of cell-matrix and cell-cell interactions on cell migration, mimic cell migration during wound healing in vivo and are compatible with imaging of live cells during migration to monitor intracellular events if desired. Besides monitoring migration of homogenous cell populations, this method has also been adopted to measure migration of individual cells in the leading edge of the scratch. Not taking into account the time for transfection of cells, in vitro scratch assay per se usually takes from several hours to overnight.
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The wound healing assay is one of the most frequently used methods for studying cell migration.
“The wound healing assay remains one of the most frequently used methods for studying cell migration [37].”
From Optimized Wound Healing Assay to Study Extracellular Vesicle-Driven Glioblastoma Cell Migration (2026) · cites this paper as [37]
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- Authors
- Liang, C.-C., Park, A. Y., & Guan, J.-L.
- Affiliations
- University of Michigan (US)
- Corresponding author
- Guan, Jun-Lin
- Venue
- Nature Protocols
- Published
- 2007-02
- Volume
- 2
- Issue
- 2
- Pages
- 329-333
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W2100592123
- Cited by
- 4,696
- References
- 5
- Citations by year
- 2026: 294 · 2025: 308 · 2024: 311 · 2023: 296 · 2022: 327 · 2021: 348 · 2020: 364 · 2019: 346 · 2018: 344 · 2017: 388 · 2016: 320 · 2015: 266 · 2014: 200 · 2013: 193 · 2012: 150
- FWCI
- 18.62
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Heart, Lung, and Blood Institute (HL 73394, R01HL073394)
- National Institute of General Medical Sciences (R01GM048050, GM-48050)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
- Health Sciences › Medicine › Rehabilitation › Wound Healing and Treatments
- Keywords
- cell migration, cell-matrix interactions, cell-cell interactions, wound healing, live-cell imaging, single-cell migration, transfection
- MeSH terms
- Cell Movement, Cytological Techniques, Image Interpretation, Computer-Assisted, Microscopy, Fluorescence, Microscopy, Phase-Contrast, Staining and Labeling, In Vitro Techniques
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