Movies 1 and 2 demonstrate protein redistribution in Jurkat T cells plated on antibody-coated stimulatory chambers.

Movie 1. ZAP-70-EGFP equilibrates between apparently static signaling assemblies. Jurkat T cells expressing ZAP-70-EGFP were plated on an antibody-coated stimulatory chamber. Beginning 2 min after plating, one image was collected every 2.5 s at the plane of the coverslip using the Zeiss LSM 510. These images were compiled into a movie that spans 2 min and plays at 20× normal speed. The elapsed time is displayed in seconds. Twenty seconds into the run, the region enclosed in the red box was photobleached. Recovery of ZAP-70-EGFP fluorescence in the bleached region is complete within 10 to 15 s.

Movie 2. The dynamic redistribution of SLP-76-EYFP upon T cell activation. SLP-76-deficient Jurkat T cells reconstituted with a SLP-76-EYFP chimera were plated on an antibody-coated stimulatory chamber. Every 4 s five images were collected along the z axis at 0.5-mm intervals using the Perkin-Elmer Ultraview spinning wheel system. At each time point, the well-focused images were projected into a single image. These images were compiled into a movie, which spans 6 min and plays at 40× normal speed.

Citation:S. C. Bunnell, V. A. Barr, C. L. Fuller, L. E. Samelson, High-resolution multicolor imaging of dynamic signaling complexes in T cells stimulated by planar substrates. Sci. STKE 2003, pl8 (2003).

These movies are viewable using Quicktime, which can be downloaded from Apple's Quicktime site.