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Boas, D. A.

Publications and source records attributed to Boas, D. A..

At least 19 recordsLinked to original sources

The Walking Brain: Mapping feature-specific cortical responses to naturally unfolding sensory experience in the real world

Understanding how the brain processes the everyday world is fundamental, yet laboratory paradigms exclude self-generated movement, unscripted events, and a continuously evolving multisensory environment. Wearable neuroimaging enables real-world measurement, but linking neural activity to uncontrolled sensory information remains challenging. Here, we extend a feature-based framework from naturalistic movie viewing to everyday behavior using wearable whole-head high-density fNIRS and participant-specific audiovisual recordings during movies and outdoor walking with no explicit task or scripted audiovisual sequence. Luminance- and sound-related cortical maps showed significant spatial correspondence, particularly in visual and auditory regions, whereas face-related correspondence was weaker. Luminance- and face-related maps were reproducible within individuals across repeated walks. Additional exploratory analysis demonstrated, to our knowledge, the first out-of-sample encoding of continuous environmental features during unconstrained real-world fNIRS, predicting cortical activity during held-out walking periods. Together, these findings establish a feature-based framework for linking cortical activity to naturally unfolding sensory experience in everyday behavior.

neuroscience↗

Evaluating Brain Flow Index from a temple-worn wearable against depth-resolved time-domain NIRS during head-down tilt and postural transitions in healthy young men

Significance: Wearable optical sensors could make continuous cerebral hemodynamic monitoring practical in daily life. Because any surface head sensor also samples extracerebral tissue, demonstrating cerebral relevance requires a reference that separates deep from superficial hemodynamics. Aim: We evaluated whether the Temple Brain Flow Index (Temple-BF), derived from a temple-worn photoplethysmographic wearable, covaried more strongly with the deeper (brain-assigned) than the superficial (scalp-assigned) hemodynamics recovered from time-domain near-infrared spectroscopy (TD-NIRS) during controlled postural challenges. Approach: Sixty sessions from 44 healthy young adult men were analyzed across three physiological challenges: 30{degrees} head-down tilt, stand-to-squat, and stand-to-supine transitions. Optical density and mean time of flight from a three-module TD-NIRS system were inverted with a Monte Carlo-derived two-layer model to obtain scalp- and brain-layer hemoglobin time series. Temple-BF was compared with these model-derived signals using lag-adjusted and zero-lag correlations, paired brain-versus-scalp comparisons, heart rate and short-separation-HbO adjusted partial correlations. Results: Median lag-adjusted correlations between Temple-BF and brain-layer {Delta}HbO traces were 0.912, 0.843, and 0.839 for head-down tilt, stand-to-squat, and stand-to-supine transitions, respectively; corresponding scalp-layer medians were 0.494, 0.700, and 0.767. Paired brain-minus-scalp differences were significant for head-down tilt (median 0.333, p < 0.001) and stand-to-squat (0.110, p < 0.001) but not for stand-to-supine (0.036, p = 0.082) transitions. Zero-lag brain-layer medians were 0.826, 0.701, and 0.796, and the association persisted after adjustment for heart rate (partial r = 0.832, 0.850, 0.654) and for short-separation superficial HbO (0.649, 0.570, 0.438). Transition-window correlations had medians of 0.878, 0.861, and 0.876, with directionally concordant transition responses in 59 of 60 sessions. Conclusions: Temple-BF tracked model-derived brain-layer TD-NIRS {Delta}HbO across all three postural challenges, with larger median correlations against the brain-layer than the scalp compartment in all three protocols, and transition responses in the same direction for almost all sessions. These results support Temple-BF as a relative marker of cerebral hemodynamic change during postural perturbations. Establishing cerebral specificity at the temple, and excluding residual systemic and superficial contributions, will require flow-sensitive references and fuller systemic monitoring.

neuroscience↗

Visual gamma stimulation causes prolonged enhancement of low-frequency blood flow oscillations across cortical regions in mice

IntroductionGamma entrainment using sensory stimuli (GENUS) uses 40Hz-pulsed sensory stimuli to entrain neural activity in the gamma band (30-150Hz). However, the effect of GENUS on low-frequency vascular oscillations has not been fully explored. ObjectivesThe objective of this study is to elucidate the effect of GENUS on vasomotion in healthy mice and potential confounds for future application in disease studies. MethodsHead-fixed, awake C57Bl/6 mice (n=18; 9M 9F) aged between 18 to 60 weeks were subjected to white light of either 40Hz visual flicker (GENUS), or constant stimulus (control). Blood flow was imaged using laser speckle contrast imaging (LSCI) before, during, immediately after 1 hour of stimulus, and 30min after the stimulus termination. ResultsA linear mixed-effects model showed that GENUS enhanced the magnitude of 0.2-0.4Hz blood flow oscillations by 38% during stimulation and by 30% at 30 minutes after stimulation compared to control when controlled for age, sex, and other factors. The effect on vasomotion was distributed across many cortical regions not limited to visual areas and lasted beyond 24 hours post-stimulus. ConclusionThese results support the exploration of GENUS for increasing vasomotion in therapeutic contexts. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=186 SRC="FIGDIR/small/729102v1_ufig1.gif" ALT="Figure 1"> View larger version (67K): org.highwire.dtl.DTLVardef@1deee2eorg.highwire.dtl.DTLVardef@e71833org.highwire.dtl.DTLVardef@1e5c4f0org.highwire.dtl.DTLVardef@1e4832e_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Decoding Spatial Attention in the Cocktail Party Problem Using Wearable Whole-head High-Density fNIRS

Spatial attention is critical for solving the cocktail party problem, a longstanding problem in neuroscience and artificial speech recognition. The ability to decode where humans are attending in a cocktail party like scene would empower applications in brain computer interfaces and assistive devices such as hearing aids. Here we demonstrate that, in an overt attention task, the attended spatial location can be decoded robustly from single trial hemodynamic responses, using a wearable whole head high density fNIRS system. We also identify critical brain regions that make the highest contribution to decoding accuracy. Specifically, we find that decoding based on a small fraction of channels within the left and right inferior parietal lobule (IPL), achieve maximal decoding accuracy comparable to all channels. These results open the way for the design of novel BCIs and assistive devices integrated with fNIRS, that can be steered by spatial attention.

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Mapping Slow Speckle Dynamics to Probe Cellular Metabolic Activity In Vivo using Laser Speckle Contrast Imaging

SignificanceLaser speckle contrast imaging (LSCI) is widely used to measure blood flow, but speckle fluctuations may also encode biologically meaningful dynamics beyond perfusion. Foundational studies in dynamic light scattering (DLS) and micro-optical coherence tomography (OCT) have also demonstrated that slow coherent signal fluctuations can arise from energy-dependent intracellular motion in in vitro and ex vivo systems. Building upon these advances, recent work has shown that LSCI has the potential to detect slow speckle dynamics (SSD) correlated with cellular dynamics in vivo. However, the biophysical mechanisms underlying SSD in intact brain tissues remain insufficiently validated. Establishing a mechanistic bridge from controlled ex vivo and in vitro conditions to in vivo brain measurements is critical for translating speckle-based imaging beyond perfusion measurements to enable label-free assessment of cellular and metabolic activity in disease models. AimThe objective of this study is to investigate the biophysical origin of the SSD in vivo and evaluate its sensitivity to intracellular metabolic activity in brain tissue. ApproachWe utilize an epi-illumination LSCI system to measure speckle contrast as a function of camera exposure time and extract characteristic decorrelation time constants. SSD was investigated in acute mouse brain slices, where blood flow is absent, to eliminate vascular confounds. Cellular metabolism was systematically modulated using 2-deoxyglucose and glucose. Complementary in vivo measurements were performed to reveal SSDs response to hyperoxia and normoxia after ischemic stroke. ResultsSSD signals persisted in acute brain slices in the absence of blood flow. Inhibition of glycolysis significantly reduced SSD, while restoration of metabolic substrates partially recovered the signal. In in vivo measurements, SSD increased during hyperoxia compared to normoxia after ischemic stroke, suggesting increased oxygen-supported cellular metabolic activity. ConclusionsThese results indicate that SSD is sensitive to energy-dependent cellular processes closely tied to metabolic activity. SSD represents a previously uncharacterized, label-free in vivo optical contrast that enables assessment of cellular metabolic activity as well as vascular dynamics. This work establishes a mechanistic foundation for using SSD as a general optical marker of cellular viability in in vivo measurements.

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Biophysical simulations of fMRI responses using realistic microvascular models: insights into distinct hemodynamics in humans and mice

1Functional Magnetic Resonance Imaging (fMRI) is broadly used to measure human brain activity, however the hemodynamic changes that comprise the fMRI response to neuronal activity are often interpreted using microscopy data in mice. These microscopy data provide ground-truth observations of how individual blood vessels respond to neuronal activity and thus form the basis of our fundamental understanding of neurovascular coupling. Although these invasive experiments provide invaluable insight, there are striking differences in the vascular architecture of mouse and human brains that may influence the hemodynamic response. Motivated by this, we developed a biophysical modeling framework for realistic hemodynamic simulations in both mouse and human cerebral cortex. For this, we utilized Vascular Anatomical Network (VAN) models that explicitly represent the full microvascular tree as a single connected network, originally based on anatomical reconstructions from a given location of mouse cerebral cortex. We extended the VAN modeling framework using synthetic VAN models representing the microvascular network at a single location of the human cerebral cortex. To account for larger size and complexity of the human VAN models, we developed an efficient computational framework to simulate the full hemodynamic responses in this human model and compared the simulated fMRI responses between mice and humans. Our biophysical simulations are based entirely on first principles (e.g., conservation of mass); model parameter values were fixed across all simulations, not tuned to fit data, as they represent meaningful physical constants taken from previous measurements. Only two simple calibrations were tuned for each simulation, to match baseline perfusion rates (blood flow) and oxygen extraction (OEF). Our results show that differences in microvasculature indeed influenced the hemodynamic response and led to observable differences in timing--e.g., the simulated fMRI response peak in humans was delayed by [~]2 s compared to that of mice, consistent with prior fMRI observations. While there are many known differences in vascular architecture in rodents and humans, we also discovered that, unexpectedly, an asymmetry in the numbers of branches of the penetrating intracortical arterioles and venules appears to be conserved across species. We demonstrate through further simulations that this anatomical property may also be needed for suitable hemodynamic responses. Our framework thus provides a valuable tool for bridging in-vivo microscopy of microvascular dynamics to human fMRI.

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High-Density Multi-Distance fNIRS Enhances Detection of Brain Activity during a Word-Color Stroop Task

STRUCTURED ABSTRACTO_ST_ABSSignificanceC_ST_ABSFunctional Near-Infrared Spectroscopy (fNIRS) enables neuroimaging in scenarios where other modalities are less suitable, such as during motion tasks or in low-resource environments. Sparse fNIRS arrays with 30mm channel spacing are widely used but have limited spatial resolution. High-density (HD) arrays with overlapping, multi-distance channels improve sensitivity and localization but increase costs and setup times. A statistical comparison of HD and sparse arrays is needed for evaluating the benefits and trade-offs of HD arrays. AimThis study provides a statistical comparison of HD and sparse fNIRS performance to inform array selection in future research. ApproachWe measured prefrontal cortex (PFC) activation during congruent and incongruent Word-Color Stroop (WCS) tasks using both Sparse and HD arrays for 17 healthy adult participants, comparing dorsolateral PFC channel and image results at the group level. ResultsWhile both arrays detected activation in channel space during incongruent WCS, channel and image space results demonstrated superior localization and sensitivity with the HD array for all WCS. ConclusionsSparse channel data may suitably detect activation from cognitively demanding tasks, like incongruent WCS. However, the HD array outperformed Sparse in detecting and localizing brain activity in image space, particularly during lower cognitive load tasks, making them more suitable for neuroimaging applications.

bioengineering↗

Mapping human cerebral blood flow with high-density, multi-channel speckle contrast optical spectroscopy

Recently, speckle contrast optical spectroscopy (SCOS) enabled non-invasive, high signal-to-noise-ratio (SNR) human cerebral blood flow (CBF) measurements, relevant for both neuroscience and clinical monitoring of diseases with CBF dysregulation. Single channel SCOS measurements limit the information obtained to only one location on the head. In this work, we develop a multi-channel SCOS system to map spatial heterogeneity in CBF changes during human brain activation. Using a galvanometer, we temporally multiplexed a free-space laser to 7 source fibers positioned at different locations on the head. Diffuse light collected from the tissue is captured by fiber bundles projecting to 17 complementary metal-oxide semiconductor (CMOS) cameras, resulting in 50 source-detector channels measuring optical density (OD) and relative CBF changes covering an area of 7.6 cm by 6.6 cm on the head. We validated the spatial specificity and stability of the system using a liquid flow phantom. We then measured brain activity during a word-color Stroop task in 15 subjects and obtained brain activation maps. The average signal changes in the channel showing the largest activation were 1.7 x 10-2 in {Delta}OD and 6.6% in CBF.

bioengineering↗

Inclusivity in fNIRS Studies: Quantifying the Impact of Hair and Skin Characteristics on Signal Quality with Practical Recommendations for Improvement

Functional Near-Infrared Spectroscopy (fNIRS) holds transformative potential for research and clinical applications in neuroscience due to its non-invasive nature and adaptability to real-world settings. However, despite its promise, fNIRS signal quality is sensitive to individual differences in biophysical factors such as hair and skin characteristics, which can significantly impact the absorption and scattering of near-infrared light. If not properly addressed, these factors risk biasing fNIRS research by disproportionately affecting signal quality across diverse populations. Our results quantify the impact of various hair properties, skin pigmentation as well as head size, sex and age on signal quality, providing quantitative guidance for future hardware advances and methodological standards to help overcome these critical barriers to inclusivity in fNIRS studies. We provide actionable guidelines for fNIRS researchers, including a suggested metadata table and recommendations for cap and optode configurations, hair management techniques, and strategies to optimize data collection across varied participants. This research paves the way for the development of more inclusive fNIRS technologies, fostering broader applicability and improved interpretability of neuroimaging data in diverse populations.

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Optical coherence tomography enables longitudinal evaluation of cell graft-directed remodeling in stroke lesions

Stem cell grafting can promote glial repair of adult stroke injuries during the subacute wound healing phase, but graft survival and glial repair outcomes are perturbed by lesion severity and mode of injury. To better understand how stroke lesion environments alter the functions of cell grafts, we employed optical coherence tomography (OCT) to longitudinally image mouse cortical photothrombotic ischemic strokes treated with allogeneic neural progenitor cell (NPC) grafts. OCT angiography, signal intensity, and signal decay resulting from optical scattering were assessed at multiple timepoints across two weeks in mice receiving an NPC graft or an injection of saline at two days after stroke. OCT scattering information revealed pronounced axial lesion contraction that naturally occurred throughout the subacute wound healing phase that was not modified by either NPC or saline treatment. By analyzing OCT signal intensity along the coronal plane, we observed dramatic contraction of the cortex away from the imaging window in the first week after stroke which impaired conventional OCT angiography but which enabled the detection of NPC graft-induced glial repair. There was moderate, but variable, NPC graft survival at photothrombotic strokes at two weeks which was inversely correlated with acute stroke lesion sizes as measured by OCT prior to treatment, suggesting a prognostic role for OCT imaging and reinforcing the dominant effect of lesion size and severity on graft outcome. Overall, our findings demonstrate the utility of OCT imaging for both tracking and predicting natural and treatment-directed changes in ischemic stroke lesion cores.

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Neurovascular Impulse Response Function (IRF) during spontaneous activity differentially reflects intrinsic neuromodulation across cortical regions

Ascending neuromodulatory projections from deep brain nuclei generate internal brain states that differentially engage specific neuronal cell types. Because neurovascular coupling is cell-type specific and neuromodulatory transmitters have vasoactive properties, we hypothesized that the impulse response function (IRF) linking spontaneous neuronal activity with hemodynamics would depend on neuromodulation. To test this hypothesis, we used optical imaging to measure (1) release of neuromodulatory transmitters norepinephrine (NE) or acetylcholine (ACh), (2) Ca2+ activity of local cortical neurons, and (3) changes in hemoglobin concentration and oxygenation across the dorsal surface of cerebral cortex during spontaneous neuronal activity in awake mice. A canonical convolution model with a global, stationary IRF (i.e., the convolution kernel) describing evolution of total hemoglobin (HbT, reflective of dilation dynamics) with respect to Ca2+, resulted in a poor fit to the data. However, the HbT time-course was well predicted, pixel-by-pixel, by a weighted sum of Ca2+ and NE time-courses. Consistent with this result, modeling HbT as a weighted sum of stationary Ca2+ - and NE-specific IRFs (IRFCa2+ and IRFNE) convolved with the respective time-courses dramatically improved the fit compared to the global IRF. IRFCa2+ and IRFNE, estimated from the data, were positive and negative, respectively. In contrast to NE, ACh was largely redundant with Ca2+ and therefore did not improve HbT estimation. Because NE covaried with arousal, we observed instances of the diminished hemodynamic coherence between cortical regions during high arousal despite coherent behavior of the underlying neuronal Ca2+ activity. We conclude that while neurovascular coupling with respect to neuronal Ca2+ is a dynamic and seemingly complex phenomenon, hemodynamic fluctuations can be captured by a simple linear model with stationary IRFs with respect to the underlying dilatory and constrictive forces. In the current study, these forces were captured by the positive IRFCa2+ (dilation) and negative IRFNE (constriction). Without accounting for NE neuromodulation and the associated vasoconstriction, diminished hemodynamic coherence, commonly referred to as "functional (dys)connectivity" in BOLD fMRI studies, can be falsely interpreted as neuronal desynchronizations.

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Carbonic anhydrase inhibitors prevent presymptomatic capillary flow disturbances in a model of cerebral amyloidosis

Structured abstractO_ST_ABSINTRODUCTIONC_ST_ABSDisturbances in microvascular flow dynamics are hypothesized to precede the symptomatic phase of Alzheimers disease (AD). However, evidence in presymptomatic AD remains elusive, underscoring the need for therapies targeting these early vascular changes. METHODSWe employed a multimodal approach, combining in vivo optical imaging, molecular techniques, and ex vivo MRI, to investigate early capillary dysfunction in Tg-SwDI mice without memory impairment. We also assessed the efficacy of carbonic anhydrase inhibitors (CAIs) in preventing capillary flow disturbances. RESULTSOur study revealed capillary flow disturbances associated with alterations in capillary morphology, adhesion molecule expression, and Amyloid-{beta} (A{beta}) load in 9-10-month-old Tg-SwDI mice without memory impairment. CAI treatment ameliorated these capillary flow disturbances, enhanced oxygen availability, and reduced A{beta} load. DISCUSSIONThese findings underscore the importance of capillary flow disturbances as early biomarkers in presymptomatic AD and highlight the potential of CAIs for preserving vascular integrity in the early stages of AD.

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Speckle contrast optical spectroscopy improves cuffless blood pressure estimation compared to photoplethysmography

Continuous and non-invasive blood pressure (BP) monitoring has the potential to greatly improve hypertension diagnosis and management, along with enabling valuable personal health monitoring in the population at large. Existing methods rely on cuff-based sphygmomanometers, which are cumbersome and disrupt sleep. Despite extensive research utilizing photoplethysmography (PPG), which is integrated into many consumer wearables, errors in continuous BP estimation are generally considered unacceptable for general use. We developed a high-speed (390 Hz) Speckle Contrast Optical Spectroscopy (SCOS) system to measure the cardiac blood flow waveform simultaneously with the PPG signal at high temporal resolution. The system utilized high speed multiplexed detection of optical speckle patterns on the wrist and finger, enabling the extraction of novel features related to BP. In comparison to PPG alone, SCOS demonstrated a notable 31% improvement (p = 3.45 * 10-7) in systolic BP estimation when integrated into subject-specific machine-learning models. The resulting errors were remarkably low (systolic BP: 0.06+/- 2.88 mmHg, diastolic BP: 0.09 +/-2.14 mmHg) across a wide range of BP variations (range SBP: 89-284 mmHg). Importantly, this improvement was sustained several weeks later within a re-measured cohort, indicating highly robust BP predictions. Looking ahead, the use of SCOS for blood flow measurements holds the potential to substantially enhance BP estimations compared to conventional PPG-based methods.

bioengineering↗

ninjaCap: A fully customizable and 3D printable headgear for fNIRS and EEG brain imaging

SignificanceAccurate sensor placement is vital for non-invasive brain imaging, particularly for functional near infrared spectroscopy (fNIRS) and diffuse optical tomography (DOT), which lack standardized layouts like EEG. Custom, manually prepared probe layouts on textile caps are often imprecise and labor-intensive. AimWe introduce a method for creating personalized, 3D-printed headgear, enabling accurate translation of 3D brain coordinates to 2D printable panels for custom fNIRS and EEG sensor layouts, reducing costs and manual labor. ApproachOur approach uses atlas-based or subject-specific head models and a spring-relaxation algorithm for flattening 3D coordinates onto 2D panels, using 10-5 EEG coordinates for reference. This process ensures geometrical fidelity, crucial for accurate probe placement. Probe geometries and holder types are customizable and printed directly on the cap, making the approach agnostic to instrument manufacturers and probe types. ResultsOur ninjaCap method offers 2.2{+/-}1.5 mm probe placement accuracy. Over the last five years, we have developed and validated this approach with over 50 cap models and 500 participants. A cloud-based ninjaCap generation pipeline along with detailed instructions is now available at openfnirs.org. ConclusionsThe ninjaCap marks a significant advancement in creating individualized neuroimaging caps, reducing costs and labor while improving probe placement accuracy, thereby reducing variability in research.

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fNIRS Dataset During Complex Scene Analysis

When analyzing complex scenes, humans often focus their attention on an object at a particular spatial location. The ability to decode the attended spatial location would facilitate brain computer interfaces for complex scene analysis (CSA). Here, we investigated capability of functional near-infrared spectroscopy (fNIRS) to decode audio-visual spatial attention in the presence of competing stimuli from multiple locations. We targeted dorsal frontoparietal network including frontal eye field (FEF) and intra-parietal sulcus (IPS) as well as superior temporal gyrus/planum temporal (STG/PT). They all were shown in previous functional magnetic resonance imaging (fMRI) studies to be activated by auditory, visual, or audio-visual spatial tasks. To date, fNIRS has not been applied to decode auditory and visual-spatial attention during CSA, and thus, no such dataset exists yet. This report provides an open-access fNIRS dataset that can be used to develop, test, and compare machine learning algorithms for classifying attended locations based on the fNIRS signals on a single trial basis.

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Targeted micro-fiber arrays for measuring and manipulating localized multi-scale neural dynamics over large, deep brain volumes during behavior

Neural population dynamics relevant for behavior vary over multiple spatial and temporal scales across 3-dimensional volumes. Current optical approaches lack the spatial coverage and resolution necessary to measure and manipulate naturally occurring patterns of large-scale, distributed dynamics within and across deep brain regions such as the striatum. We designed a new micro-fiber array and imaging approach capable of chronically measuring and optogenetically manipulating local dynamics across over 100 targeted locations simultaneously in head-fixed and freely moving mice. We developed a semi-automated micro-CT based strategy to precisely localize positions of each optical fiber. This highly-customizable approach enables investigation of multi-scale spatial and temporal patterns of cell-type and neurotransmitter specific signals over arbitrary 3-D volumes at a spatial resolution and coverage previously inaccessible. We applied this method to resolve rapid dopamine release dynamics across the striatum volume which revealed distinct, modality specific spatiotemporal patterns in response to salient sensory stimuli extending over millimeters of tissue. Targeted optogenetics through our fiber arrays enabled flexible control of neural signaling on multiple spatial scales, better matching endogenous signaling patterns, and spatial localization of behavioral function across large circuits.

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Widefield in vivo imaging system with two fluorescence and two reflectance channels, a single sCMOS detector, and shielded illumination

SignificanceWidefield microscopy of the entire dorsal part of mouse cerebral cortex enables large-scale ("mesoscopic") imaging of different aspects of neuronal activity with spectrally compatible fluorescent indicators as well as hemodynamics via oxy- and deoxyhemoglobin absorption. Versatile and cost-effective imaging systems are needed for large-scale, color-multiplexed imaging of multiple fluorescent and intrinsic contrasts. AimDevelop a system for mesoscopic imaging of two fluorescent and two reflectance channels. ApproachExcitation of red and green fluorescence is achieved through epi-illumination. Hemoglobin absorption imaging is achieved using 525- and 625-nm LEDs positioned around the objective lens. An aluminum hemisphere placed between objective and cranial window provides diffuse illumination of the brain. Signals are recorded sequentially by a single sCMOS detector. ResultsWe demonstrate performance of our imaging system by recording large-scale spontaneous and stimulus-evoked neuronal, cholinergic, and hemodynamic activity in awake head-fixed mice with a curved "crystal skull" window expressing the red calcium indicator jRGECO1a and the green acetylcholine sensor GRABACh3.0. Shielding of illumination light through the aluminum hemisphere enables concurrent recording of pupil diameter changes. ConclusionsOur widefield microscope design with single camera can be used to acquire multiple aspects of brain physiology and is compatible with behavioral readouts of pupil diameter.

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Multi-Scale Label-free Human Brain Imaging with Integrated Serial Sectioning Polarization Sensitive Optical Coherence Tomography and Two-Photon Microscopy

The study of neurodegenerative processes in the human brain requires a comprehensive understanding of cytoarchitectonic, myeloarchitectonic, and vascular structures. Recent computational advances have enabled volumetric reconstruction of the human brain using thousands of stained slices, however, tissue distortions and loss resulting from standard histological processing have hindered deformation-free reconstruction of the human brain. The development of a multi-scale and volumetric human brain imaging technique that can measure intact brain structure would be a major technical advance. Here, we describe the development of integrated serial sectioning Polarization Sensitive Optical Coherence Tomography (PSOCT) and Two Photon Microscopy (2PM) to provide label-free multi-contrast imaging, including scattering, birefringence and autofluorescence of human brain tissue. We demonstrate that high-throughput reconstruction of 4x4x2cm3 sample blocks and simple registration of PSOCT and 2PM images enable comprehensive analysis of myelin content, vascular structure, and cellular information. We show that 2 m in-plane resolution 2PM images provide microscopic validation and enrichment of the cellular information provided by the PSOCT optical property maps on the same sample, revealing the sophisticated capillary networks and lipofuscin filled cell bodies across the cortical layers. Our method is applicable to the study of a variety of pathological processes, including demyelination, cell loss, and microvascular changes in neurodegenerative diseases such as Alzheimers disease (AD) and Chronic Traumatic Encephalopathy (CTE).

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