Adaptive optical methods for in vivo imaging in developing Zebra fish

The humble Zebra fish is rapidly establishing itself as the model of choice for a wide range of biological investigations, in particular at the developing embryo stage. Single Plane Illumination Microscopy (SPIM) has already been shown to be a very powerful method with which to image such samples over extended periods of time. However, the sample is only around 2mm long and its physical structure heavily influenced by the beating heart with the circulation of blood causing movement throughout the fish on the scale of several microns. The paper reports on the development of an active synchronization method, which optically “freezes” motion and significantly reduces light induced toxicity and bleaching within the sample. This has been combined with adaptive optics to remove both instrument and sample induced aberrations enabling live high resolution imaging within the beating heart with micron resolution over extended time periods.