High-speed confocal fluorescence imaging with a novel line scanning microscope.

Research in the life sciences increasingly involves the investigation of fast dynamic processes at the cellular and subcellular level. It requires tools to image complex systems with high temporal resolution in three-dimensional space. For this task, we introduce the concept of a fast fluorescence line scanner providing image acquisition speeds in excess of 100 frames per second at 512 x 512 pixels.

Rapid three-dimensional imaging and analysis of the beating embryonic heart reveals functional changes during development.

We report an accurate method for studying the functional dynamics of the beating embryonic zebrafish heart. The fast cardiac contraction rate and the high velocity of blood cells have made it difficult to study cellular and subcellular events relating to heart function in vivo. We have devised a dynamic three-dimensional acquisition, reconstruction, and analysis procedure by combining (1) a newly developed confocal slit-scanning microscope, (2) novel strategies for collecting and synchronizing cyclic image sequences to build volumes with high temporal and spatial resolution over the entire depth of the beating heart, and (3) data analysis and reduction protocols for the systematic extraction of quantitative information to describe phenotype and function.