Neurophotonics Visibility, Indexing, and Impact

Quantitative phase microscopy (QPM) has recently emerged as a new powerful quantitative imaging technique well suited to noninvasively explore a transparent specimen with a nanometric axial sensitivity. In this review, we expose the recent developments of quantitative phase-digital holographic microscopy (QP-DHM). Quantitative phase-digital holographic microscopy (QP-DHM) represents an important and efficient quantitative phase method to explore cell structure and dynamics. In a second part, the most relevant QPM applications in the field of cell biology are summarized. A particular emphasis is placed on the original biological information, which can be derived from the quantitative phase signal. In a third part, recent applications obtained, with QP-DHM in the field of cellular neuroscience, namely the possibility to optically resolve neuronal network activity and spine dynamics, are presented. Furthermore, potential applications of QPM related to psychiatry through the identification of new and original cell biomarkers that, when combined with a range of other biomarkers, could significantly contribute to the determination of high risk developmental trajectories for psychiatric disorders, are discussed.

This study investigates the correspondence of the cortical sensitivity of electroencephalography (EEG) and near-infrared spectroscopy (NIRS). EEG forward model sensitivity to the cerebral cortex was calculated for 329 EEG electrodes following the 10-5 EEG positioning system using a segmented structural magnetic resonance imaging scan of a human subject. NIRS forward model sensitivity was calculated for the same subject using 156 NIRS source-detector pairs selected from 32 source and 32 detector optodes positioned on the scalp using a subset of the 10-5 EEG positioning system. Sensitivity correlations between colocalized NIRS source-detector pair groups and EEG channels yielded R=0.46±0.08 . Groups of NIRS source-detector pairs with maximum correlations to EEG electrode sensitivities are tabulated. The mean correlation between the point spread functions for EEG and NIRS regions of interest (ROI) was R=0.43±0.07 . Spherical ROIs with radii of 26 mm yielded the maximum correlation between EEG and NIRS averaged across all cortical mesh nodes. These sensitivity correlations between EEG and NIRS should be taken into account when designing multimodal studies of neurovascular coupling and when using NIRS as a statistical prior for EEG source localization.

Three-dimensional tissue cultures have been used as effective models for studying different diseases, including epilepsy. High-throughput, nondestructive techniques are essential for rapid assessment of disease-related processes, such as progressive cell death. An ultrahigh-resolution optical coherence microscopy (UHR-OCM) system with ∼1.5  μm axial resolution and ∼2.3  μm transverse resolution was developed to evaluate seizure-induced neuronal injury in organotypic rat hippocampal cultures. The capability of UHR-OCM to visualize cells in neural tissue was confirmed by comparison of UHR-OCM images with confocal immunostained images of the same cultures. In order to evaluate the progression of neuronal injury, UHR-OCM images were obtained from cultures on 7, 14, 21, and 28 days in vitro (DIVs). In comparison to DIV 7, statistically significant reductions in three-dimensional cell count and culture thickness from UHR-OCM images were observed on subsequent time points. In cultures treated with kynurenic acid, significantly less reduction in cell count and culture thickness was observed compared to the control specimens. These results demonstrate the capability of UHR-OCM to perform rapid, label-free, and nondestructive evaluation of neuronal death in organotypic hippocampal cultures. UHR-OCM, in combination with three-dimensional tissue cultures, can potentially prove to be a promising tool for high-throughput screening of drugs targeting various disorders.

While near-infrared spectroscopy (NIRS) has been increasingly applied to neuroimaging and functional connectivity studies in infants, it has not been quantitatively examined as to what extent the deep tissue (such as cerebral tissue) as opposed to shallow tissue (such as scalp), contributes to NIRS signals measured in infants. A method for separating the effects of deep- and shallow-tissue layers was applied to data of nine sleeping three-month-old infants who had been exposed to 3-s speech sounds or silence (i.e., resting state) and whose hemodynamic changes over their bilateral temporal cortices had been measured by using an NIRS system with multiple source-detector (S-D) distances. The deep-layer contribution was found to be large during resting [67% at S-D 20 mm, 78% at S-D 30 mm for oxygenated hemoglobin (oxy-Hb)] as well as during the speech condition (72% at S-D 20 mm, 82% at S-D 30 mm for oxy-Hb). A left-right connectivity analysis showed that correlation coefficients between left and right channels did not differ between original- and deep-layer signals under no-stimulus conditions and that of original- and deep-layer signals were larger than those of the shallow layer. These results suggest that NIRS signals obtained in infants with appropriate S-D distances largely reflected cerebral hemodynamic changes.

Cerebral edema develops in response to a variety of conditions, including traumatic brain injury and stroke, and contributes to the poor prognosis associated with these injuries. This study examines the use of optical coherence tomography (OCT) for detecting cerebral edema in vivo. Three-dimensional imaging of an in vivo water intoxication model in mice was performed using a spectral-domain OCT system centered at 1300 nm. The change in attenuation coefficient was calculated and cerebral blood flow was analyzed using Doppler OCT techniques. We found that the average attenuation coefficient in the cerebral cortex decreased over time as edema progressed. The initial decrease began within minutes of inducing cerebral edema and a maximum decrease of 8% was observed by the end of the experiment. Additionally, cerebral blood flow slowed during late-stage edema. Analysis of local regions revealed the same trend at various locations in the brain, consistent with the global nature of the cerebral edema model used in this study. These results demonstrate that OCT is capable of detecting in vivo optical changes occurring due to cerebral edema and highlights the potential of OCT for precise spatiotemporal detection of cerebral edema.

There has been a rapid rise in the number of publications using functional near infrared spectroscopy (fNIRS) for human developmental research over the past decade. However test–retest reliability of this measure of brain activation in infants remains unknown. To assess this, we utilized data from a longitudinal cohort who participated in an fNIRS study on social perception at two age points. Thirteen infants had valid data from two sessions held 8.5 months apart (4 to 8 months and 12 to 16 months). Inter- and intrasession fNIRS test–retest reliability was assessed at the individual and group levels using the oxyhemoglobin (HbO₂) signal. Infant compliance with the study was similar in both sessions (assessed by the proportion of time infants looked to the stimuli), and there was minimal discrepancy in sensor placement over the targeted area between sessions. At the group level, good spatial overlap of significant responses and signal reliability was seen (spatial overlap was 0.941 and average signal change within an region of interest was r=0.896). At participant level, spatial overlap was acceptable (<0.5 on average across infants) although signal reliability varied between participants. This first study of test–retest reliability of fNIRS in infants shows encouraging results, particularly for group-based analysis.

Functional near-infrared spectroscopy (fNIRS) is becoming a popular tool in developmental neuroscience for mapping functional localized brain responses. However, as it cannot provide information about underlying anatomy, researchers have begun to conduct spatial registration of fNIRS channels to cortical anatomy in adults. The current work investigated this issue with infants by coregistering fNIRS and magnetic resonance imaging (MRI) data from 55 individuals. Our findings suggest that fNIRS channels can be reliably registered with regions in the frontal and temporal cortex of infants from 4 to 7 months of age. Although some macro-anatomical regions are difficult to consistently define, others are more stable and fNIRS channels on an age-appropriate MRI template are often consistent with individual infant MRIs. We have generated a standardized scalp surface map of fNIRS channel locators to reliably locate cortical regions for fNIRS developmental researchers. This new map can be used to identify the inferior frontal gyrus, superior temporal sulcus (STS) region [which includes the superior and middle temporal gyri (MTG) nearest to the STS], and MTG and temporal-parietal regions in 4- to 7-month-old infants. Future work will model data for the whole head, taking into account the properties of light transport in tissue, and expanding to different ages across development.

The current study aimed to explore the neural substrate for atomoxetine effects on attentional control in school-aged children with attention deficit hyperactivity disorder (ADHD) using functional near-infrared spectroscopy (fNIRS), which can be applied to young children with ADHD more easily than conventional neuroimaging modalities. Using fNIRS, we monitored the oxy-hemoglobin signal changes of 15 ADHD children (6 to 14 years old) performing an oddball task before and 1.5 h after atomoxetine or placebo administration, in a randomized, double-blind, placebo-controlled, crossover design. Fifteen age-, gender-, and intelligence quotient-matched normal controls without atomoxetine administration were also monitored. In the control subjects, the oddball task recruited the right prefrontal and inferior parietal cortices. The right prefrontal and parietal activation was normalized after atomoxetine administration in ADHD children. This was in contrast to our previous study using a similar protocol showing methylphenidate-induced normalization of only the right prefrontal function. fNIRS allows the detection of differential neuropharmacological profiles of both substances in the attentional network: the neuropharmacological effects of atomoxetine to upregulate the noradrenergic system reflected in the right prefrontal and inferior parietal activations and those of methylphenidate to upregulate the dopamine system reflected in the prefrontal cortex activation.

A major goal of the BRAIN Initiative is the development of technologies to monitor neuronal network activity during active information processing. Toward this goal, genetically encoded calcium indicator proteins have become widely used for reporting activity in preparations ranging from invertebrates to awake mammals. However, slow response times, the narrow sensitivity range of Ca²+ and in some cases, poor signal-to-noise ratio still limit their usefulness. Here, we review recent improvements in the field of neural activity-sensitive probe design with a focus on the GCaMP family of calcium indicator proteins. In this context, we present our newly developed Fast-GCaMPs, which have up to 4-fold accelerated off-responses compared with the next-fastest GCaMP, GCaMP6f. Fast-GCaMPs were designed by destabilizing the association of the hydrophobic pocket of calcium-bound calmodulin with the RS20 binding domain, an intramolecular interaction that protects the green fluorescent protein chromophore. Fast-GCaMP6f-RS06 and Fast-GCaMP6f-RS09 have rapid off-responses in stopped-flow fluorimetry, in neocortical brain slices, and in the intact cerebellum in vivo. Fast-GCaMP6f variants should be useful for tracking action potentials closely spaced in time, and for following neural activity in fast-changing compartments, such as axons and dendrites. Finally, we discuss strategies that may allow tracking of a wider range of neuronal firing rates and improve spike detection.

Recent studies have demonstrated functional near-infrared spectroscopy (fNIRS) to be a viable and sensitive method for imaging sensorimotor cortex activity in children with cerebral palsy (CP). However, during unilateral finger tapping, children with CP often exhibit unintended motions in the nontapping hand, known as mirror motions, which confuse the interpretation of resulting fNIRS images. This work presents a method for separating some of the mirror motion contributions to fNIRS images and demonstrates its application to fNIRS data from four children with CP performing a finger-tapping task with mirror motions. Finger motion and arm muscle activity were measured simultaneously with fNIRS signals using motion tracking and electromyography (EMG), respectively. Subsequently, subject-specific regressors were created from the motion capture or EMG data and independent component analysis was combined with a general linear model to create an fNIRS image representing activation due to the tapping hand and one image representing activation due to the mirror hand. The proposed method can provide information on how mirror motions contribute to fNIRS images, and in some cases, it helps remove mirror motion contamination from the tapping hand activation images.