High-speed imaging in fluids

High-speed imaging is in popular demand for a broad range of experiments in fluids. It allows for a detailed visualization of the event under study by acquiring a series of image frames captured at high temporal and spatial resolution. This review covers high-speed imaging basics, by defining criteria for high-speed imaging experiments in fluids and to give rule-of-thumbs for a series of cases. It also considers stroboscopic imaging, triggering and illumination, and scaling issues. It provides guidelines for testing and calibration. Ultra high-speed imaging at frame rates exceeding 1 million frames per second is reviewed, and the combination of conventional experiments in fluids techniques with high-speed imaging techniques are discussed. The review is concluded with a high-speed imaging chart, which summarizes criteria for temporal scale and spatial scale and which facilitates the selection of a high-speed imaging system for the applicatio

High-resolution 3D optical microscopy inside the beating zebrafish heart using prospective optical gating

3D fluorescence imaging is a fundamental tool in the study of functional and developmental biology, but effective imaging is particularly difficult in moving structures such as the beating heart. We have developed a non-invasive real-time optical gating system that is able to exploit the periodic nature of the motion to acquire high resolution 3D images of the normally-beating zebrafish heart without any unnecessary exposure of the sample to harmful excitation light. In order for the image stack to be artefact-free, it is essential to use a synchronization source that is invariant as the sample is scanned in 3D. We therefore describe a scheme whereby fluorescence image slices are scanned through the sample while a brightfield camera sharing the same objective lens is maintained at a fixed focus, with correction of sample drift also included. This enables us to maintain, throughout an extended 3D volume, the same standard of synchronization we have previously demonstrated in and near a single 2D plane. Thus we are able image the complete beating zebrafish heart exactly as if the heart had been artificially stopped, but sidestepping this undesirable interference with the heart and instead allowing the heart to beat as normal