Home Failure Case Library Scratch is too wide or uneven
Wound Healing Assay moderate

Scratch is too wide or uneven

Symptom
Scratch edges are rough and the width varies along the line, making it impossible to accurately compare migration distances later.
Common Causes
  1. 1 Pressure on the pipette tip is uneven
  2. 2 Different scratch tools used across wells
  3. 3 Monolayer confluence is not uniform
Solutions
  1. 1 Use the same model of pipette tip — or better, a dedicated scratch tool
  2. 2 Move in a single straight line at constant speed
  3. 3 Plate evenly and confirm consistent monolayer before scratching
Related Video (2)
YouTube (Curated Tutorials) ★ 85
Scratch Assay Tutorial
"Direct scratch assay tutorial addressing the core technique and likely covering proper pipette tip pressure control for consistent scratch creation"
YouTube (Curated Tutorials) ★ 80
Wound healing
"Wound healing assay setup and analysis video demonstrating correct execution of the scratch step to avoid the uneven width problem"
Source: xiaohongshu.com ↗
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