Technologies for imaging neural activity in large volumes

Neural circuitry has evolved to form distributed networks that act dynamically across large volumes. Collecting data from individual planes, conventional microscopy cannot sample circuitry across large volumes at the temporal resolution relevant to neural circuit function and behaviors. Here, we review emerging technologies for rapid volume imaging of neural circuitry. We focus on two critical challenges: the inertia of optical systems, which limits image speed, and aberrations, which restrict the image volume. Optical sampling time must be long enough to ensure high-fidelity measurements, but optimized sampling strategies and point spread function engineering can facilitate rapid volume imaging of neural activity within this constraint. We also discuss new computational strategies for the processing and analysis of volume imaging data of increasing size and complexity. Together, optical and computational advances are providing a broader view of neural circuit dynamics, and help elucidate how brain regions work in concert to support behavior

The Refinement of Ipsilateral Eye Retinotopic Maps Is Increased by Removing the Dominant Contralateral Eye in Adult Mice

Background: Shortly after eye opening, initially disorganized visual cortex circuitry is rapidly refined to form smooth retinotopic maps. This process asymptotes long before adulthood, but it is unknown whether further refinement is possible. Prior work from our lab has shown that the retinotopic map of the non-dominant ipsilateral eye develops faster when the dominant contralateral eye is removed. We examined whether input from the contralateral eye might also limit the ultimate refinement of the ipsilateral eye retinotopic map in adults. In addition, we examined whether the increased refinement involved the recruitment of adjacent cortical area. Methodology/Principal Findings: By surgically implanting a chronic optical window over visual cortex in mice, we repeatedly measured the degree of retinotopic map refinement using quantitative intrinsic signal optical imaging over four weeks. We removed the contralateral eye and observed that the retinotopic map for the ipsilateral eye was further refined and the maximum magnitude of response increased. However, these changes were not accompanied by an increase in the area of responsive cortex. Conclusions/Significance: Since the retinotopic map was functionally refined to a greater degree without taking over adjacent cortical area, we conclude that input from the contralateral eye limits the normal refinement of visual cortica

Sustainability science graduate students as boundary spanners

Graduate training in sustainability science (SS) focuses on interdisciplinary research, stakeholder-researcher partnerships, and creating solutions from knowledge. But becoming a sustainability scientist also requires specialized training that addresses the complex boundaries implicit in sustainability science approaches to solving social-ecological system challenges. Using boundary spanning as a framework, we use a case study of the Sustainability Solutions Initiative (SSI) at the University of Maine to explicate key elements for graduate education training in SS. We used a mixed-methods approach, including a quantitative survey and autoethnographic reflection, to analyze our experiences as SSI doctoral students. Through this research, we identified four essential SS boundaries that build on core sustainability competencies which need to be addressed in SS graduate programs, including: disciplines within academia, students and their advisors, researchers and stakeholders, and place-based and generalizable research. We identified key elements of training necessary to help students understand and navigate these boundaries using core competencies. We then offer six best practice recommendations to provide a basis for a SS education framework. Our reflections are intended for academic leaders in SS who are training new scientists to solve complex sustainability challenges. Our experiences as a cohort of doctoral students with diverse academic and professional backgrounds provide a unique opportunity to reflect not only on the challenges of SS but also on the specific needs of students and programs striving to provide solutions

Wide field-of-view, multi-region, two-photon imaging of neuronal activity in the mammalian brain

Two-photon calcium imaging provides an optical readout of neuronal activity in populations of neurons with subcellular resolution. However, conventional two-photon imaging systems are limited in their field of view to ~1 mm2, precluding the visualization of multiple cortical areas simultaneously. Here, we demonstrate a two-photon microscope with an expanded field of view (>9.5 mm2) for rapidly reconfigurable simultaneous scanning of widely separated populations of neurons. We custom designed and assembled an optimized scan engine, objective, and two independently positionable, temporally multiplexed excitation pathways. We used this new microscope to measure activity correlations between two cortical visual areas in mice during visual processing

A touchscreen based global motion perception task for mice

Global motion perception is a function of higher, or extrastriate, visual system circuitry. These circuits can be engaged in visually driven navigation, a behavior at which mice are adept. However, the properties of global motion perception in mice are unclear. Therefore, we developed a touchscreen-based, two-alternative forced choice (2AFC) task to explore global motion detection in mice using random dot kinematograms (RDK). Performance data was used to compute coherence thresholds for global motion perception. The touchscreen-based task allowed for parallel training and testing with multiple chambers and minimal experimenter intervention with mice performing hundreds of trials per session. Parameters of the random dot kinematograms, including dot size, lifetime, and speed, were tested. Mice learned to discriminate kinematograms whose median motion direction differed by 90 degrees in 7-24days after a 10-14day pre-training period. The average coherence threshold (measured at 70% correct) in mice for this task was 22±5%, with a dot diameter of 3.88mm and speed of 58.2mm/s. Our results confirm the ability of mice to perform global motion discriminations, and the touchscreen assay provides a flexible, automated, and relatively high throughput method with which to probe complex visual function in mice

Stream-dependent development of higher visual cortical areas

Multiple cortical areas contribute to visual processing in mice. However, the functional organization and development of higher visual areas are unclear. Here, we used intrinsic signal optical imaging and 2-photon calcium imaging to map visual responses in adult and developing mice. We found that visually driven activity was well-correlated among higher visual areas within two distinct subnetworks resembling the dorsal and ventral visual streams. Visual response magnitude in dorsal stream areas slowly increased over the first two weeks of visual experience. By contrast, ventral stream areas exhibited strong responses shortly after eye opening. Neurons in a dorsal stream area showed little change in their tuning sharpness to oriented gratings while those in a ventral stream area increased stimulus selectivity and expanded their receptive fields significantly. Together, these findings provide a functional basis for grouping subnetworks of mouse visual areas and revealed stream differences in the development of receptive field properties

Improving data quality in neuronal population recordings

Understanding how the brain operates requires understanding how large sets of neurons function together. Modern recording technology makes it possible to simultaneously record the activity of hundreds of neurons, and technological developments will soon allow recording of thousands or tens of thousands. As with all experimental techniques, these methods are subject to confounds that complicate the interpretation of such recordings, and could lead to erroneous scientific conclusions. Here, we discuss methods for assessing and improving the quality of data from these techniques, and outline likely future directions in this field