Abstract
We introduce a scanless optical method to image neuronal activity in three dimensions simultaneously. Using a spatial light modulator and a custom-designed phase mask, we illuminate and collect light simultaneously from different focal planes and perform calcium imaging of neuronal activity in vitro and in vivo. This method, combining structured illumination with volume projection imaging, could be used as a technological platform for brain activity mapping.
Highlights
Optical imaging of neural activity has several advantages over alternative strategies such as patch electrodes or electrode arrays
From the very first microscope designed by Leeuwenhoek, image acquisition is typically limited to a single plane, while most biological structures are threedimensional, requiring sequential scanning for volumetric imaging
We present an alternative to traditional scanning-based imaging by taking advantage of programmable three-dimensional illumination with spatial light modulators (SLM) to simultaneously excite neurons located at different focal points, together with a wavefront coded imaging approach that enables us to collect light from all focal points simultaneously
Summary
Optical imaging of neural activity has several advantages over alternative strategies such as patch electrodes or electrode arrays. It is minimally invasive and allows for the monitoring of large ensembles of neurons with single-cell resolution (Yuste and Katz, 1991). It is compatible with a large variety of functional sensors (voltage, calcium or metabolic indicators, either exogenous or genetically encoded), and can be used for chronic imaging of defined cell populations in vitro or in vivo (Grynkiewicz et al, 1985; Kralj et al, 2012; Chen et al, 2013). Our technique supersedes scanning by illuminating and collects light from the neurons of interest in parallel
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