Prospective gating for 3D imaging of the beating zebrafish heart in embryonic development studies

We demonstrate the use of prospective gating from continuously acquired brightfield images of zebrafish embryos to trigger the acquisition of fluorescence images with the heart at a precisely selected position in its cycle. The laser exposure of the sample is reduced by an order of magnitude compared to alternative techniques which acquire many separate fluorescence images for each section before selecting the most appropriate ones to build up a consistent 3D image stack. We present results obtained using our SPIM system including 3D reconstructions of the living, beating heart, acquired using optical gating without the need for any pharmacological or electrophysiological intervention, and discuss possible wider applications of our technique