Neurophotonics Editor in Chief Anna Devor welcomes an expanded editorial board and announces three Special Sections for the coming year.

Significance: Quantitative measures of blood flow and metabolism are essential for improved assessment of brain health and response to ischemic injury.

Aim: We demonstrate a multimodal technique for measuring the cerebral metabolic rate of oxygen (CMRO₂) in the rodent brain on an absolute scale (μM O₂  /  min).

Approach: We use laser speckle imaging at 809 nm and spatial frequency domain imaging at 655, 730, and 850 nm to obtain spatiotemporal maps of cerebral blood flow, tissue absorption (μ_a), and tissue scattering (μs^′). Knowledge of these three values enables calculation of a characteristic blood flow speed, which in turn is input to a mathematical model with a “zero-flow” boundary condition to calculate absolute CMRO₂. We apply this method to a rat model of cardiac arrest (CA) and cardiopulmonary resuscitation. With this model, the zero-flow condition occurs during entry into CA.

Results: The CMRO₂ values calculated with our method are in good agreement with those measured with magnetic resonance and positron emission tomography by other groups.

Conclusions: Our technique provides a quantitative metric of absolute cerebral metabolism that can potentially be used for comparison between animals and longitudinal monitoring of a single animal over multiple days. Though this report focuses on metabolism in a model of ischemia and reperfusion, this technique can potentially be applied to far broader types of acute brain injury and whole-body pathological occurrences.

Significance: High-density diffuse optical tomography (HD-DOT) has been shown to approach the resolution and localization accuracy of blood oxygen level dependent-functional magnetic resonance imaging in the adult brain by exploiting densely spaced, overlapping samples of the probed tissue volume, but the technique has to date required large and cumbersome optical fiber arrays.

Aim: To evaluate a wearable HD-DOT system that provides a comparable sampling density to large, fiber-based HD-DOT systems, but with vastly improved ergonomics.

Approach: We investigated the performance of this system by replicating a series of classic visual stimulation paradigms, carried out in one highly sampled participant during 15 sessions to assess imaging performance and repeatability.

Results: Hemodynamic response functions and cortical activation maps replicate the results obtained with larger fiber-based systems. Our results demonstrate focal activations in both oxyhemoglobin and deoxyhemoglobin with a high degree of repeatability observed across all sessions. A comparison with a simulated low-density array explicitly demonstrates the improvements in spatial localization, resolution, repeatability, and image contrast that can be obtained with this high-density technology.

Conclusions: The system offers the possibility for minimally constrained, spatially resolved functional imaging of the human brain in almost any environment and holds particular promise in enabling neuroscience applications outside of the laboratory setting. It also opens up new opportunities to investigate populations unsuited to traditional imaging technologies.

Significance: Light-sheet fluorescence microscopy (LSFM) is a powerful technique for highspeed volumetric functional imaging. However, in typical light-sheet microscopes, the illumination and collection optics impose significant constraints upon the imaging of non-transparent brain tissues. We demonstrate that these constraints can be surmounted using a new class of implantable photonic neural probes.

Aim: Mass manufacturable, silicon-based light-sheet photonic neural probes can generate planar patterned illumination at arbitrary depths in brain tissues without any additional micro-optic components.

Approach: We develop implantable photonic neural probes that generate light sheets in tissue. The probes were fabricated in a photonics foundry on 200-mm-diameter silicon wafers. The light sheets were characterized in fluorescein and in free space. The probe-enabled imaging approach was tested in fixed, in vitro, and in vivo mouse brain tissues. Imaging tests were also performed using fluorescent beads suspended in agarose.

Results: The probes had 5 to 10 addressable sheets and average sheet thicknesses <16 μm for propagation distances up to 300 μm in free space. Imaging areas were as large as ≈240 μm × 490 μm in brain tissue. Image contrast was enhanced relative to epifluorescence microscopy. .

Conclusions: The neural probes can lead to new variants of LSFM for deep brain imaging and experiments in freely moving animals.

Significance: To explore brain architecture and pathology, a consistent and reliable methodology to visualize the three-dimensional cerebral microvasculature is beneficial. Perfusion-based vascular labeling is quick and easily deliverable. However, the quality of vascular labeling can vary with perfusion-based labels due to aggregate formation, leakage, rapid photobleaching, and incomplete perfusion.

Aim: We describe a simple, two-day protocol combining perfusion-based labeling with a two-day clearing step that facilitates whole-brain, three-dimensional microvascular imaging and characterization.

Approach: The combination of retro-orbital injection of Lectin-Dylight-649 to label the vasculature, the clearing process of a modified iDISCO+ protocol, and light-sheet imaging collectively enables a comprehensive view of the cerebrovasculature.

Results: We observed ∼threefold increase in contrast-to-background ratio of Lectin-Dylight-649 vascular labeling over endogenous green fluorescent protein fluorescence from a transgenic mouse model. With light-sheet microscopy, we demonstrate sharp visualization of cerebral microvasculature throughout the intact mouse brain.

Conclusions: Our tissue preparation protocol requires fairly routine processing steps and is compatible with multiple types of optical microscopy.

Significance: We present a new optical method for modulating cortical activity in multiple locations and across multiple time points with high spatial and temporal precision. Our method uses infrared light and does not require dyes or transgenic modifications. It is compatible with a number of other stimulation and recording techniques.

Aim: Infrared neural stimulation (INS) has been largely confined to single point stimuli. In this study, we expand upon this approach and develop a rapidly switched fiber array capable of generation of stimulus patterns. Our prototype is capable of stimulating at nine separate locations but is easily scalable.

Approach: Our device is made of commercially available components: a solid-state infrared laser, a piezoelectric fiber coupled optical switch, and 200-μm diameter optical fibers. We validate it using intrinsic optical signal imaging of INS responses in macaque and squirrel monkey sensory cortical areas.

Results: We demonstrate that our switched array can consistently generate responses in primate cortex, consistent with earlier single channel INS investigations.

Conclusions: Our device can successfully target the cortical surface, either at one specific region or multiple points spread out across different areas. It is compatible with a host of other imaging and stimulation modalities.

Significance: Diffuse correlation spectroscopy (DCS) is an emerging noninvasive, diffuse optical modality that purportedly enables direct measurements of microvasculature blood flow. Functional optical coherence tomography angiography (OCT-A) can resolve blood flow in vessels as fine as capillaries and thus has the capability to validate key attributes of the DCS signal.

Aim: To characterize activity in cortical vasculature within the spatial volume that is probed by DCS and to identify populations of blood vessels that are most representative of the DCS signals.

Approach: We performed simultaneous measurements of somatosensory-evoked cerebral blood flow in mice in vivo using both DCS and OCT-A.

Results: We resolved sensory-evoked blood flow in the somatosensory cortex with both modalities. Vessels with diameters smaller than 10  μm featured higher peak flow rates during the initial poststimulus positive increase in flow, whereas larger vessels exhibited considerably larger magnitude of the subsequent undershoot. The simultaneously recorded DCS waveforms correlated most highly with flow in the smallest vessels, yet featured a more prominent undershoot.

Conclusions: Our direct, multiscale, multimodal cross-validation measurements of functional blood flow support the assertion that the DCS signal preferentially represents flow in microvasculature. The significantly greater undershoot in DCS, however, suggests a more spatially complex relationship to flow in cortical vasculature during functional activation.

Significance: There is an emerging need for convenient and continuous bedside monitoring of full-term newborns with hypoxic-ischemic brain damage (HIBD) to determine whether early intervention is required. Functional near-infrared spectroscopy (fNIRS)-based resting-state brain network analysis, which could provide an effective evaluation method, remains to be extensively studied.

Aim: Our study aims to verify the feasibility of fNIRS-based resting-state brain networks for evaluating brain function in infants with HIBD to provide a new and effective means for clinical research in neonatal HIBD.

Approach: Thirteen neonates with HIBD were scanned using fNIRS in the resting state. The brain network properties were explored to attempt to extract effective features as recognition indicators.

Results: Compared with healthy controls, newborns with HIBD showed decreased brain functional connectivity. Specifically, there were severe losses of long-range functional connectivity of the contralateral parietal-temporal lobe, contralateral parietal-frontal lobe, and contralateral parietal lobe. The node degree showed a widespread decrease in the left frontal middle gyrus, left superior frontal gyrus dorsal, and right central posterior gyrus. However, newborns with HIBD showed a significantly higher local network efficiency (*p  <  0.05). Subsequently, network indicators based on small-worldness, local efficiency, modularity, and normalized clustering coefficient were extracted for HIBD identification with the accuracy observed as 79.17%.

Conclusions: Our findings indicate that fNIRS-based resting-state brain network analysis could support early HIBD diagnosis.

Significance: Functional near-infrared spectroscopy (fNIRS) is an increasingly popular tool in auditory research, but the range of analysis procedures employed across studies may complicate the interpretation of data.

Aim: We aim to assess the impact of different analysis procedures on the morphology, detection, and lateralization of auditory responses in fNIRS. Specifically, we determine whether averaging or generalized linear model (GLM)-based analysis generates different experimental conclusions when applied to a block-protocol design. The impact of parameter selection of GLMs on detecting auditory-evoked responses was also quantified.

Approach: 17 listeners were exposed to three commonly employed auditory stimuli: noise, speech, and silence. A block design, comprising sounds of 5 s duration and 10 to 20 s silent intervals, was employed.

Results: Both analysis procedures generated similar response morphologies and amplitude estimates, and both indicated that responses to speech were significantly greater than to noise or silence. Neither approach indicated a significant effect of brain hemisphere on responses to speech. Methods to correct for systemic hemodynamic responses using short channels improved detection at the individual level.

Conclusions: Consistent with theoretical considerations, simulations, and other experimental domains, GLM and averaging analyses generate the same group-level experimental conclusions. We release this dataset publicly for use in future development and optimization of algorithms.

Significance: The establishment of a light propagation analysis-based scalp-cortex correlation (SCC) between the scalp location of the source–detector (SD) pair and brain regions is essential for measuring functional brain development in the first 2 years of life using functional near-infrared spectroscopy (fNIRS).

Aim: We aimed to reveal the optics-based SCC of 0-, 1-, and 2-year-olds (yo) and the suitable SD distance for this age period.

Approach: Light propagation analyses using age-appropriate head models were conducted on SD pairs at 10-10 fiducial points on the scalp to obtain optics-based SCC and its metrics: the number of corresponding brain regions (N_CBR), selectivity and sensitivity of the most likely corresponding brain region (MLCBR), and consistency of the MLCBR across developmental ages. Moreover, we assessed the suitable SD distances for 0-, 1-, and 2-yo by simultaneously considering the selectivity and sensitivity of the MLCBR.

Results: Age-related changes in the SCC metrics were observed. For instance, the N_CBR of 0-yo was larger than that of 1- and 2-yo. Conversely, the selectivity of 0-yo was lower than that of 1- and 2-yo. The sensitivity of 1-yo was higher than that of 0-yo at 15- to 30-mm SD distances and higher than that of 2-yo at 10-mm SD distance. Notably, the MLCBR of the fiducial points around the longitudinal fissure was inconsistent across age groups. An SD distance between 15 and 25 mm was found to be appropriate for satisfying both sensitivity and selectivity requirements. In addition, this work provides reference tables of optics-based SCC for 0-, 1-, and 2-yo.

Conclusions: Optics-based SCC will be informative in designing and explaining child developmental studies using fNIRS. The suitable SD distances were between 15 and 25 mm for the first 2 years of life.

Significance: Image reconstruction of fNIRS data is a useful technique for transforming channel-based fNIRS into a volumetric representation and managing spatial variance based on optode location. We present an innovative integrated pipeline for image reconstruction of fNIRS data using either MRI templates or individual anatomy.

Aim: We demonstrate a pipeline with accompanying code to allow users to clean and prepare optode location information, prepare and standardize individual anatomical images, create the light model, run the 3D image reconstruction, and analyze data in group space.

Approach: We synthesize a combination of new and existing software packages to create a complete pipeline, from raw data to analysis.

Results: This pipeline has been tested using both templates and individual anatomy, and on data from different fNIRS data collection systems. We show high temporal correlations between channel-based and image-based fNIRS data. In addition, we demonstrate the reliability of this pipeline with a sample dataset that included 74 children as part of a longitudinal study taking place in Scotland. We demonstrate good correspondence between data in channel space and image reconstructed data.

Conclusions: The pipeline presented here makes a unique contribution by integrating multiple tools to assemble a complete pipeline for image reconstruction in fNIRS. We highlight further issues that may be of interest to future software developers in the field.

Significance: Early monolingual versus bilingual experience induces adaptations in the development of linguistic and cognitive processes, and it modulates functional activation patterns during the first months of life. Resting-state functional connectivity (RSFC) is a convenient approach to study the functional organization of the infant brain. RSFC can be measured in infants during natural sleep, and it allows to simultaneously investigate various functional systems. Adaptations have been observed in RSFC due to a lifelong bilingual experience. Investigating whether bilingualism-induced adaptations in RSFC begin to emerge early in development has important implications for our understanding of how the infant brain’s organization can be shaped by early environmental factors.

Aims: We attempt to describe RSFC using functional near-infrared spectroscopy (fNIRS) and to examine whether it adapts to early monolingual versus bilingual environments. We also present an fNIRS data preprocessing and analysis pipeline that can be used to reliably characterize RSFC in development and to reduce false positives and flawed results interpretations.

Methods: We measured spontaneous hemodynamic brain activity in a large cohort (N  =  99) of 4-month-old monolingual and bilingual infants using fNIRS. We implemented group-level approaches based on independent component analysis to examine RSFC, while providing proper control for physiological confounds and multiple comparisons.

Results: At the group level, we describe the functional organization of the 4-month-old infant brain in large-scale cortical networks. Unbiased group-level comparisons revealed no differences in RSFC between monolingual and bilingual infants at this age.

Conclusions: High-quality fNIRS data provide a means to reliably describe RSFC patterns in the infant brain. The proposed group-level RSFC analyses allow to assess differences in RSFC across experimental conditions. An effect of early bilingual experience in RSFC was not observed, suggesting that adaptations might only emerge during explicit linguistic tasks, or at a later point in development.

Significance: Designing optode layouts is an essential step for functional near-infrared spectroscopy (fNIRS) experiments as the quality of the measured signal and the sensitivity to cortical regions-of-interest depend on how optodes are arranged on the scalp. This becomes particularly relevant for fNIRS-based brain–computer interfaces (BCIs), where developing robust systems with few optodes is crucial for clinical applications.

Aim: Available resources often dictate the approach researchers use for optode-layout design. We investigated whether guiding optode layout design using different amounts of subject-specific magnetic resonance imaging (MRI) data affects the fNIRS signal quality and sensitivity to brain activation when healthy participants perform mental-imagery tasks typically used in fNIRS-BCI experiments.

Approach: We compared four approaches that incrementally incorporated subject-specific MRI information while participants performed mental-calculation, mental-rotation, and inner-speech tasks. The literature-based approach (LIT) used a literature review to guide the optode layout design. The probabilistic approach (PROB) employed individual anatomical data and probabilistic maps of functional MRI (fMRI)-activation from an independent dataset. The individual fMRI (iFMRI) approach used individual anatomical and fMRI data, and the fourth approach used individual anatomical, functional, and vascular information of the same subject (fVASC).

Results: The four approaches resulted in different optode layouts and the more informed approaches outperformed the minimally informed approach (LIT) in terms of signal quality and sensitivity. Further, PROB, iFMRI, and fVASC approaches resulted in a similar outcome.

Conclusions: We conclude that additional individual MRI data lead to a better outcome, but that not all the modalities tested here are required to achieve a robust setup. Finally, we give preliminary advice to efficiently using resources for developing robust optode layouts for BCI and neurofeedback applications.

Significance: We propose a customized animal-specific head cap and an near-infrared spectroscopy (NIRS) system to obtain NIRS signals in mobile small animals. NIRS studies in mobile small animals provide a feasible solution for comprehensive brain function studies. Aim: We aim to develop and validate a multichannel NIRS system capable of performing functional brain imaging along with a closed-box stimulation kit for small animals in mobile conditions. Approach: The customized NIRS system uses light-weight long optical fibers, along with a customized light-weight head cap to securely attach the optical fibers to the mouse. A customized stimulation box was designed to perform various stimuli in a controlled environment. The system performance was tested in a visual stimulation task on eight anesthetized mice and eight freely moving mice. Results: Following the visual stimulation task, we observed a significant stimulation-related oxyhemoglobin (HbO) increase in the visual cortex of freely moving mice during the task. In contrast, HbO concentration did not change significantly in the visual cortex of anesthetized mice. Conclusions: We demonstrate the feasibility of a wearable, multichannel NIRS system for small animals in a less confined experimental design.