Search

Genetically encoded voltage indicators (GEVIs) enable optical recording of electrical signals in the brain, providing subthreshold sensitivity and temporal resolution not possible with calcium indicators. However, one- and two-photon voltage imaging over prolonged periods with the same GEVI has not yet been demonstrated. Here, we report engineering of ASAP family GEVIs to enhance photostability by inversion of the fluorescence-voltage relationship. Two of the resulting GEVIs, ASAP4b and ASAP4e, respond to 100-mV depolarizations with ≥180% fluorescence increases, compared with the 50% fluorescence decrease of the parental ASAP3. With standard microscopy equipment, ASAP4e enables single-trial detection of spikes in mice over the course of minutes. Unlike GEVIs previously used for one-photon voltage recordings, ASAP4b and ASAP4e also perform well under two-photon illumination. By imaging voltage and calcium simultaneously, we show that ASAP4b and ASAP4e can identify place cells and detect voltage spikes with better temporal resolution than commonly used calcium indicators. Thus, ASAP4b and ASAP4e extend the capabilities of voltage imaging to standard one- and two-photon microscopes while improving the duration of voltage recordings.

Imaging methods that span both functional measures in living tissue and anatomical measures in fixed tissue have played critical roles in advancing our understanding of the brain. However, making direct comparisons between different imaging modalities, particularly spanning living and fixed tissue, has remained challenging. For example, comparing brain-wide neural dynamics across experiments and aligning such data to anatomical resources, such as gene expression patterns or connectomes, requires precise alignment to a common set of anatomical coordinates. However, reaching this goal is difficult because registering in vivo functional imaging data to ex vivo reference atlases requires accommodating differences in imaging modality, microscope specification, and sample preparation. We overcome these challenges in Drosophila by building an in vivo reference atlas from multiphoton-imaged brains, called the Functional Drosophila Atlas. We then develop a registration pipeline, BrIdge For Registering Over Statistical Templates (BIFROST), for transforming neural imaging data into this common space and for importing ex vivo resources such as connectomes. Using genetically labeled cell types as ground truth, we demonstrate registration with a precision of less than 10 microns. Overall, BIFROST provides a pipeline for registering functional imaging datasets in the fly, both within and across experiments. [ABSTRACT FROM AUTHOR]

Optimal control of networks is to minimize the cost function of a network in a dynamical process with an optimal control strategy. For the time-invariant linear systems, $\dot {x}(t)=A x(t)+B u(t)$ , and the traditional linear quadratic regulator (LQR), which minimizes a quadratic cost function, has been well established given both the adjacency matrix $A$ and the control input matrix $B$. However, this conventional approach is not applicable when we have the freedom to design $B$. In this article, we investigate the situation when the input matrix $B$ is a variable to be designed to reduce the control cost. First, the problem is formulated and we establish an equivalent expression of the quadratic cost function with respect to $B$ , which is difficult to obtain within the traditional theoretical framework as it requires obtaining an explicit solution of a Riccati differential equation (RDE). Next, we derive the gradient of the quadratic cost function with respect to the matrix variable $B$ analytically. Further, we obtain three inequalities of the cost functions, after which several possible design (optimization) problems are discussed, and algorithms based on gradient information are proposed. It is shown that the cost of controlling the LTI systems can be significantly reduced when the input matrix becomes “designable.” We find that the nodes connected to input sources can be sparsely identified and they are distributed as evenly as possible in the LTI networks if one wants to control the networks with the lowest cost. Our findings help us better understand how the LTI systems should be controlled through designing the input matrix. [ABSTRACT FROM AUTHOR]

Optical imaging of neuronal voltage dynamics is invaluable to studying brain functions. However, high-speed imaging at significant depth is challenging due to the limitations of the short pixel dwell time and the maximum permissible excitation power in tissues. We report high-speed, deep voltage imaging by applying adaptive excitation, which illuminates the regions of interest only. We imaged neuronal voltage activities across two planes down to 500-630 μm depth in the awake mouse brain. Our techniques can be straightforwardly applied to typical two-photon microscopes., Competing Interests: Additional Declarations: There is NO Competing Interest.

The heterogeneity of brain imaging methods in neuroscience provides rich data that cannot be captured by a single technique, and our interpretations benefit from approaches that enable easy comparison both within and across different data types. For example, comparing brain-wide neural dynamics across experiments and aligning such data to anatomical resources, such as gene expression patterns or connectomes, requires precise alignment to a common set of anatomical coordinates. However, this is challenging because registering in vivo functional imaging data to ex vivo reference atlases requires accommodating differences in imaging modality, microscope specification, and sample preparation. We overcome these challenges in Drosophila by building an in vivo reference atlas from multiphoton-imaged brains, called the Functional Drosophila Atlas (FDA). We then develop a two-step pipeline, BrIdge For Registering Over Statistical Templates (BIFROST), for transforming neural imaging data into this common space and for importing ex vivo resources such as connectomes. Using genetically labeled cell types as ground truth, we demonstrate registration with a precision of less than 10 microns. Overall, BIFROST provides a pipeline for registering functional imaging datasets in the fly, both within and across experiments., Significance: Large-scale functional imaging experiments in Drosophila have given us new insights into neural activity in various sensory and behavioral contexts. However, precisely registering volumetric images from different studies has proven challenging, limiting quantitative comparisons of data across experiments. Here, we address this limitation by developing BIFROST, a registration pipeline robust to differences across experimental setups and datasets. We benchmark this pipeline by genetically labeling cell types in the fly brain and demonstrate sub-10 micron registration precision, both across specimens and across laboratories. We further demonstrate accurate registration between in-vivo brain volumes and ultrastructural connectomes, enabling direct structure-function comparisons in future experiments.