Isaac Kauvar

Unconventional Face‐On Texture and Exceptional In‐Plane Order of a High Mobility n‐Type Polymer

Substantial in-plane crystallinity and dominant face-on stacking are observed in thin films of a high-mobility n-type rylene-thiophene copolymer. Spun films of the polymer, previously thought to have little or no order are found to exhibit an ordered microstructure at both interfaces, and in the bulk. The implications of this type of packing and crystalline morphology are discussed as they relate to thin-film transistors.

SPED Light Sheet Microscopy: Fast Mapping of Biological System Structure and Function.

The goal of understanding living nervous systems has driven interest in high-speed and large field-of-view volumetric imaging at cellular resolution. Light sheet microscopy approaches have emerged for cellular-resolution functional brain imaging in small organisms such as larval zebrafish, but remain fundamentally limited in speed. Here, we have developed SPED light sheet microscopy, which combines large volumetric field-of-view via an extended depth of field with the optical sectioning of light sheet microscopy, thereby eliminating the need to physically scan detection objectives for volumetric imaging. SPED enables scanning of thousands of volumes-per-second, limited only by camera acquisition rate, through the harnessing of optical mechanisms that normally result in unwanted spherical aberrations. We demonstrate capabilities of SPED microscopy by performing fast sub-cellular resolution imaging of CLARITY mouse brains and cellular-resolution volumetric Ca(2+) imaging of entire zebrafish nervous systems. Together, SPED light sheet methods enable high-speed cellular-resolution volumetric mapping of biological system structure and function.

Simultaneous fast measurement of circuit dynamics at multiple sites across the mammalian brain

Real-time activity measurements from multiple specific cell populations and projections are likely to be important for understanding the brain as a dynamical system. Here we developed frame-projected independent-fiber photometry (FIP), which we used to record fluorescence activity signals from many brain regions simultaneously in freely behaving mice. We explored the versatility of the FIP microscope by quantifying real-time activity relationships among many brain regions during social behavior, simultaneously recording activity along multiple axonal pathways during sensory experience, performing simultaneous two-color activity recording, and applying optical perturbation tuned to elicit dynamics that match naturally occurring patterns observed during behavior.

Global Representations of Goal-Directed Behavior in Distinct Cell Types of Mouse Neocortex

SUMMARY The successful planning and execution of adaptive behaviors in mammals may require long-range coordination of neural networks throughout cerebral cortex. The neuronal implementation of signals that could orchestrate cortex-wide activity remains unclear. Here, we develop and apply methods for cortex-wide Ca2+ imaging in mice performing decision-making behavior and identify a global cortical representation of task engagement encoded in the activity dynamics of both single cells and superficial neuropil distributed across the majority of dorsal cortex. The activity of multiple molecularly defined cell types was found to reflect this representation with type-specific dynamics. Focal optogenetic inhibition tiled across cortex revealed a crucial role for frontal cortex in triggering this cortex-wide phenomenon; local inhibition of this region blocked both the cortex-wide response to task-initiating cues and the voluntary behavior. These findings reveal cell-type-specific processes in cortex for globally representing goal-directed behavior and identify a major cortical node that gates the global broadcast of task-related information.

Modulation of prefrontal cortex excitation/inhibition balance rescues social behavior in CNTNAP2-deficient mice

Using optogenetics to modulate the excitation/inhibition balance in the prefrontal cortex of an autism-like mouse model rescued social behavior deficits. Social interactions light up Neurophysiological phenomena that underlie the symptoms of autism remain unclear. Genetics-based mouse models of autism have suggested that there is an increase in the neuronal excitation/inhibition (E:I) balance. An optogenetically driven increase in this E:I balance leads to social deficits in mice. Using mice lacking CNTNAP2, a gene known to be associated with autism in humans, Selimbeyoglu and colleagues now show that real-time optogenetic modulation of the E:I balance rescued social behavior deficits and hyperactivity in these animals. This study highlights the potential for modulating neural circuits in the brain as a strategy for treating autism. Alterations in the balance between neuronal excitation and inhibition (E:I balance) have been implicated in the neural circuit activity–based processes that contribute to autism phenotypes. We investigated whether acutely reducing E:I balance in mouse brain could correct deficits in social behavior. We used mice lacking the CNTNAP2 gene, which has been implicated in autism, and achieved a temporally precise reduction in E:I balance in the medial prefrontal cortex (mPFC) either by optogenetically increasing the excitability of inhibitory parvalbumin (PV) neurons or decreasing the excitability of excitatory pyramidal neurons. Surprisingly, both of these distinct, real-time, and reversible optogenetic modulations acutely rescued deficits in social behavior and hyperactivity in adult mice lacking CNTNAP2. Using fiber photometry, we discovered that native mPFC PV neuronal activity differed between CNTNAP2 knockout and wild-type mice. During social interactions with other mice, PV neuron activity increased in wild-type mice compared to interactions with a novel object, whereas this difference was not observed in CNTNAP2 knockout mice. Together, these results suggest that real-time modulation of E:I balance in the mouse prefrontal cortex can rescue social behavior deficits reminiscent of autism phenotypes.

Ancestral Circuits for the Coordinated Modulation of Brain State

Internal states of the brain profoundly influence behavior. Fluctuating states such as alertness can be governed by neuromodulation, but the underlying mechanisms and cell types involved are not fully understood. We developed a method to globally screen for cell types involved in behavior by integrating brain-wide activity imaging with high-content molecular phenotyping and volume registration at cellular resolution. We used this method (MultiMAP) to record from 22 neuromodulatory cell types in behaving zebrafish during a reaction-time task that reports alertness. We identified multiple monoaminergic, cholinergic, and peptidergic cell types linked to alertness and found that activity in these cell types was mutually correlated during heightened alertness. We next recorded from and controlled homologous neuromodulatory cells in mice; alertness-related cell-type dynamics exhibited striking evolutionary conservation and modulated behavior similarly. These experiments establish a method for unbiased discovery of cellular elements underlying behavior and reveal an evolutionarily conserved set of diverse neuromodulatory systems that collectively govern internal state.

Extended field-of-view and increased-signal 3D holographic illumination with time-division multiplexing

Phase spatial light modulators (SLMs) are widely used for generating multifocal three-dimensional (3D) illumination patterns, but these are limited to a field of view constrained by the pixel count or size of the SLM. Further, with two-photon SLM-based excitation, increasing the number of focal spots penalizes the total signal linearly--requiring more laser power than is available or can be tolerated by the sample. Here we analyze and demonstrate a method of using galvanometer mirrors to time-sequentially reposition multiple 3D holograms, both extending the field of view and increasing the total time-averaged two-photon signal. We apply our approach to 3D two-photon in vivo neuronal calcium imaging.

Adaptive color display via perceptually-driven factored spectral projection

Fundamental display characteristics are constantly being improved, especially resolution, dynamic range, and color reproduction. However, whereas high resolution and high-dynamic range displays have matured as a technology, it remains largely unclear how to extend the color gamut of a display without either sacrificing light throughput or making other tradeoffs. In this paper, we advocate for adaptive color display; with hardware implementations that allow for color primaries to be dynamically chosen, an optimal gamut and corresponding pixel states can be computed in a content-adaptive and user-centric manner. We build a flexible gamut projector and develop a perceptually-driven optimization framework that robustly factors a wide color gamut target image into a set of time-multiplexed primaries and corresponding pixel values. We demonstrate that adaptive primary selection has many benefits over fixed gamut selection and show that our algorithm for joint primary selection and gamut mapping performs better than existing methods. Finally, we evaluate the proposed computational display system extensively in simulation and, via photographs and user experiments, with a prototype adaptive color projector.

Variable Aperture Light Field Photography: Overcoming the Diffraction-Limited Spatio-Angular Resolution Tradeoff

Light fields have many applications in machine vision, consumer photography, robotics, and microscopy. However, the prevalent resolution limits of existing light field imaging systems hinder widespread adoption. In this paper, we analyze fundamental resolution limits of light field cameras in the diffraction limit. We propose a sequential, coded-aperture-style acquisition scheme that optimizes the resolution of a light field reconstructed from multiple photographs captured from different perspectives and fnumber settings. We also show that the proposed acquisition scheme facilitates high dynamic range light field imaging and demonstrate a proof-of-concept prototype system. With this work, we hope to advance our understanding of the resolution limits of light field photography and develop practical computational imaging systems to overcome them.

Aperture interference and the volumetric resolution of light field fluorescence microscopy

Light field microscopy (LFM) is an emerging technique for volumetric fluorescence imaging, but widespread use is hampered by its poor spatial resolution. Using diffraction-based analysis we show how this degraded resolution arises because conventional LFM aims to sample four dimensions of the light field. By instead prioritizing 3D volumetric information over 4D sampling, we can optically interfere certain redundant angular samples to allow higher spatial resolution while maintaining enough angular information for depth discrimination. With this in mind, we design a number of aperture plane sampling schemes, characterize their frequency support and invertibility, and describe how their relative performance depends on the operating signal-to-noise regime. With simulations and a prototype, we demonstrate a time-sequential amplitude mask-based acquisition approach that outperforms conventional LFM in terms of both spatial resolution and axial field of view.