This PDF file contains the editorial “Three Senior Editors Join Neurophotonics at the Start of Its Third Year” for NPh Vol. 3 Issue 02

This PDF file contains the editorial “Emerging research areas in need of neurophotonics: report from the 2014 Aarhus Capillary Transit Time Heterogeneity (CTH) meeting” for NPh Vol. 3 Issue 02

The use of bioluminescent reporters in neuroscience research continues to grow at a rapid pace as their applications and unique advantages over conventional fluorescent reporters become more appreciated. Here, we describe practical methods and principles for detecting and imaging bioluminescence from live cells and animals. We systematically tested various components of our conventional fluorescence microscope to optimize it for long-term bioluminescence imaging. High-resolution bioluminescence images from live neurons were obtained with our microscope setup, which could be continuously captured for several hours with no signs of phototoxicity. Bioluminescence from the mouse brain was also imaged noninvasively through the intact skull with a conventional luminescence imager. These methods demonstrate how bioluminescence can be routinely detected and measured from live cells and animals in a cost-effective way with common reagents and equipment.

The vascular response during cortical microelectrode insertion was measured with amplitude decorrelation-based quantitative optical coherence angiography (OCA). Four different shank-style microelectrode configurations were inserted in murine motor cortex beneath a surgically implanted window in discrete steps while OCA images were collected and processed for angiography and flowmetry. Quantitative measurements included tissue displacement (measured by optical flow), perfused capillary density, and capillary flow velocity. The primary effect of insertion was mechanical perturbation, the effects of which included tissue displacement, arteriolar rupture, and compression of a branch of the anterior cerebral artery causing a global decrease in flow. Other effects observed included local flow drop-out in the region immediately surrounding the microelectrode. The mean basal capillary network velocity for all animals was 0.23 (±0.05  SD) and 0.18 (±0.07  SD) mm/s for capillaries from 100 to 300  μm and 300 to 500  μm, respectively. Upon insertion, the 2-shank electrode arrays caused a decrease in capillary flow density and velocity, while the results from other configurations were not different from controls. The proximity to large vessels appears to play a larger role than the array configuration. These results can guide neurosurgeons and electrode designers to minimize trauma and ischemia during microelectrode insertion.

Optical techniques have gained substantial interest over the past four decades for biomedical imaging due to their unique advantages, which may suggest their use as alternatives to conventional methodologies. Several optical techniques have been successfully adapted to clinical practice and biomedical research to monitor tissue structure and function in both humans and animal models. This paper reviews the analysis of the optical properties of brain tissue in the wavelength range between 500 and 1000 nm by three different diffuse optical reflectance methods: spatially modulated illumination, orthogonal diffuse light spectroscopy, and dual-wavelength laser speckle imaging, to monitor changes in brain tissue morphology, chromophore content, and metabolism following head injury. After induction of closed head injury upon anesthetized mice by weight-drop method, significant changes in hemoglobin oxygen saturation, blood flow, and metabolism were readily detectible by all three optical setups, up to 1 h post-trauma. Furthermore, the experimental results clearly demonstrate the feasibility and reliability of the three methodologies, and the differences between the system performances and capabilities are also discussed. The long-term goal of this line of study is to combine these optical systems to study brain pathophysiology in high spatiotemporal resolution using additional models of brain trauma. Such combined use of complementary algorithms should fill the gaps in each system’s capabilities, toward the development of a noninvasive, quantitative tool to expand our knowledge of the principles underlying brain function following trauma, and to monitor the efficacy of therapeutic interventions in the clinic.

Functional near-infrared spectroscopy (fNIRS) is a noninvasive neuroimaging technique used to measure changes in oxygenated hemoglobin (oxy-Hb) and deoxygenated hemoglobin (deoxy-Hb) in the brain. In this study, we present a decomposition approach based on single-channel independent component analysis (scICA) to investigate the contribution of physiological noise to fNIRS signals during rest. Single-channel ICA is an underdetermined decomposition method, which separates a single time series into components containing nonredundant spectral information. Using scICA, fNIRS signals from a total of 17 subjects were decomposed into the constituent physiological components. The percentage contribution of the classes of physiology to the fNIRS signals including low-frequency (LF) fluctuations, respiration, and cardiac oscillations was estimated using spectral domain classification methods. Our results show that LF oscillations accounted for 40% to 55% of total power of both the oxy-Hb and deoxy-Hb signals. Respiration and its harmonics accounted for 10% to 30% of the power, and cardiac pulsations and cardio-respiratory components accounted for 10% to 30%. We describe this scICA method for decomposing fNIRS signals, which unlike other approaches to spatial covariance reduction is applicable to both single- or multiple-channel fNIRS signals and discuss how this approach allows functionally distinct sources of noise with disjoint spectral support to be separated from obscuring systemic physiology.

Functional near-infrared spectroscopy has yet to be implemented as a stand-alone technique within an ophthalmology clinical setting, despite its promising advantages. The present study aims to further investigate reliability of visual cortical signals. This was achieved by: (1) assessing the effects of optode placements using the 10–20 International System of Electrode Placement consisting of 28 channels, (2) determining effects of stimulus size on response, and (3) evaluating response variability as a result of cap placement across three sessions. Ten participants with mean age 23.8±4.8 years (five male) and varying types of hair color and thickness were recruited. Visual stimuli of black-and-white checkerboards, reversing at a frequency of 7.5 Hz were presented. Visual angles of individual checker squares included 1 deg, 2 deg, 5 deg, 9 deg, and 18 deg. The number of channels that showed response was analyzed for each participant, stimulus size, and session. 1-deg stimulus showed the greatest activation. One of three data collection sessions for each participant gave different results (p<0.05). Hair color and thickness each had an effect upon the overall HbO (p<0.05), while only color had a significant effect for HbD (p<0.05). A reliable level of robustness and consistency is still required for clinical implementation and assessment of visual dysfunction.

Despite promising advantages such as low cost and portability of functional near-infrared spectroscopy (fNIRS), it has yet to be widely implemented outside of basic research. Specifically, fNIRS has yet to be proven as a standalone tool within a clinical setting. The objective of this study was to assess hemodynamic concentration changes at the primary and premotor motor cortices as a result of simple whole-hand grasping and sequential finger-opposition (tapping) tasks. These tasks were repeated over 3 days in a randomized manner. Ten healthy young adults (23.8±4.8  years) participated in the study. Quantitatively, no statistically significant differences were discovered between the levels of activation for the two motor tasks (p<0.05). Overall, the signals were consistent across all 3 days. The findings show that both finger-opposition and hand grasping can be used interchangeably in fNIRS for assessment of motor function which would be useful in further advancing techniques for clinical implementation.