3D time lapse imaging and quantitative analysis of multilayered Endothelial cells migration

Yokogawa CSU (confocal scanner unit) is a confocal unit using a microlens-enhanced dual Nipkow disk confocal optical system, which has been shown to be less harmful to living cells compared to conventional single beam scanning devices. The CQ1 is an all-in-one confocal quantitative imaging cytometer based on the CSU. Here we report the 3D time lapse live cell imaging in a multilayered cell sheet using CQ1.

Methods:

• Five-layered myoblast cell sheets were constructed from human skeletal muscle myoblasts (HSMM) and human skeletal muscle fibroblasts (HSMF)
• HSMMs and HSMFs were labeled with CellTrackerTM Orange
• Human umbilical vein endothelial cells (HUVEC) expressing GFP (GFP-HUVEC) were overlaid by the cell sheet and co-cultured.
• Time lapse imaging (67 hours, 30 min interval, 40x objective lens , 49 fields) was performed by CQ1 equipped with an internal incubation chamber to regulate culture environment.

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