Fast volume-scanning light sheet microscopy reveals transient neuronal events
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Light sheet fluorescence microscopy offers considerable potential to the cellular neuroscience community as it makes it possible to image extensive areas of neuronal structures, such as axons or dendrites, with a low light budget, thereby minimizing phototoxicity. However, the shallow depth of a light sheet, which is critical for achieving high contrast, well resolved images, adds a significant challenge if fast functional imaging is also required, as multiple images need to be collected across several image planes. Consequently, fast functional imaging of neurons is typically restricted to a small tissue volume where part of the neuronal structure lies within the plane of a single image. Here we describe a method by which fast functional imaging can be achieved across a much larger tissue volume; a custom-built light sheet microscope is presented that includes a synchronized galvo mirror and electrically tunable lens, enabling high speed acquisition of images across a configurable depth. We assess the utility of this technique by acquiring fast functional Ca2+ imaging data across a neuron’s dendritic arbour in mammalian brain tissue.
Haslehurst , P , Yang , Z , Dholakia , K & Emptage , N 2018 , ' Fast volume-scanning light sheet microscopy reveals transient neuronal events ' Biomedical Optics Express , vol 9 , no. 5 , pp. 2154-2167 . DOI: 10.1364/BOE.9.002154
Biomedical Optics Express
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DescriptionFunding: UK EPSRC EP/P030017/1.
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