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What is Cell Migration / 2D scratch assay? And What you need to know about it?

  • Writer: CLYTE research team
    CLYTE research team
  • Mar 5
  • 3 min read

Updated: May 25

Different types of Scratch Assays
Image By: Senior, J.J. (2024). Cell Migration in Wound Healing. In: Brüning-Richardson, A., Knipp, S. (eds) Cell Migration in Development, Health and Disease. Learning Materials in Biosciences. Springer, Cham. https://doi.org/10.1007/978-3-031-64532-7_3

Cell migration is a fundamental process in various physiological and pathological contexts, including wound healing, immune responses, and cancer metastasis. To study this phenomenon in vitro, researchers frequently use the scratch assay, also known as the wound healing assay. This method provides valuable insights into cell movement mechanisms and has applications across multiple scientific disciplines, including cancer research, pharmacology, tissue engineering, and developmental biology.


What is a Scratch Assay?

A scratch assay is a widely used technique to assess cell migration in laboratory conditions. It involves creating a "scratch" or cell-free gap in a confluent monolayer of cells grown on a substrate. This artificial wound mimics tissue damage, prompting cells at the edges to migrate into the gap. Researchers can monitor and quantify this migration under different conditions to analyze cellular behavior. The scratch assay is particularly useful for studying collective cell migration, where groups of cells move in coordination.


Applications of Scratch Assays in Research and Industry

Scratch assays are widely utilized in multiple scientific fields, including:

1. Cancer Research

  • Evaluating the invasive potential of cancer cells.

  • Testing the efficacy of anti-metastatic drugs.

2. Pharmacology

  • Screening compounds that influence cell motility.

  • Supporting drug development for tissue repair and cancer treatment.

3. Tissue Engineering

  • Studying cell-material interactions.

  • Optimizing biocompatible scaffold designs for tissue regeneration.

4. Developmental Biology

  • Investigating cell migration during embryonic development.

  • Understanding genetic and molecular influences on cell movement.


Best Practices for Conducting a Scratch Assay

To ensure accurate and reproducible results in scratch assays, follow these best practices:

✅ Achieve Full Confluency

  • Ensure 100% confluence before creating the scratch for consistent results.

✅ Use Standardized Scratching Tools

  • Manual scratching with pipette tips can cause inconsistencies. Use automated or standardized scratch devices for uniform width and edges.

✅ Pre-Wash Cells

  • Rinse cells with phosphate-buffered saline (PBS) before scratching to remove debris and prevent irregular migration patterns.

✅ Maintain Optimal Culture Conditions

  • Perform assays under controlled temperature, humidity, and CO₂ levels to ensure cell health.

✅ Use Serum-Free Media

  • Minimize cell proliferation interference by using serum-free media when studying pure migration effects.


Common Mistakes and How to Avoid Them

Even experienced researchers face challenges with scratch assays. Avoid these common pitfalls:

❌ Inconsistent Scratch Creation – Manual scratching leads to variability in wound size and shape. Use precise tools like CellCut for standardization.

❌ Damaging Adjacent Cells – Applying too much pressure can injure surrounding cells, altering their behavior.

❌ Uneven Cell Density – Inconsistent cell seeding leads to unreliable migration data.

❌ Lack of Controls – Always include untreated wells or known migration inhibitors for accurate comparisons.


Scratch Assay tool demo
Researcher demonstrating the use of Project CellCut v3 prototype, for scratch assays.

Revolutionizing Scratch Assays with CellCut

What is CellCut?

To address the challenges of manual scratch assays, we developed CellCut, an innovative 3D-printed device designed to standardize scratch assays.

By positioning CellCut over a multi-well plate and performing a single motion, researchers can create consistent scratches across all wells—eliminating variability and improving experimental reliability.


Key Benefits of CellCut:

✔ Enhanced Reproducibility – Standardized scratches reduce data variability.✔ Increased Throughput – Scratch multiple wells simultaneously, saving time.✔ Improved Accuracy – Uniform wound areas enable precise migration assessments.✔ Reduced Workload – Simplifies the scratch assay process, making it more efficient and reliable.


Try CellCut Today!

Interested in testing CellCut prototypes? Sign up for early access by visiting our CLYTE Partnership page or clicking the Get Started button at the top of this page.


🔬 Standardize your scratch assays with CellCut – the future of cell migration research!


References



CytCut 3.0

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