To quantify MC cluster formation in Ubow mice, ear skin whole mounts were imaged using a LSM780 (Zeiss) confocal microscope equipped with a Plan-Apochromat ×20 M27 objective (Zeiss). Z stacks of 25 to 42 µm with a 2-µm step size and multiple tiles were acquired using ZEN black software, and 2 × 2 tiled images (1.2 × 1.2 mm; covering a minimum of 600 MCs) were stitched using ZEN blue software. Statistical analysis and quantification of spatial MC distribution were performed using ClusterQuant2D software21,60. Detailed information of the analysis is provided in Supplementary Note 5.
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.