FIGURE 1.

Imaging of intimal Ca²⁺ signals in open human artery segments (A) Opened artery segments mounted on silicone inserts were loaded with Ca²⁺ indicator (Cal-520 AM) and imaged using a spinning disk confocal microscope (B) Representative images (top) show maximal intensity projections (cumulative fluorescence over 60s) of an original recording and the Ca²⁺ signals detected along the vessel intima (monochromatic mask). A continuous time-lapse tracing was generated by tracking event centers over the sampling period; individual image panels show single-frame snapshots of Ca²⁺ signals at the time points indicated in the tracing (dotted lines) (C) A zoomed pseudo-color image in the sampled intimal field shows two distinct Ca²⁺ events (dotted boxes). The boxed regions (below) are compressed along the y-axis and extrapolated over time (D) Nuclear staining was performed after Ca²⁺ imaging to assess confluency of endothelial cells (vertically oriented nuclei) along the intima; image to the right shows boxed region. Bar is 50 μm; EC, endothelial cell; VSMC, vascular smooth muscle cell.