Intrinsic Optical Signals Research Articles

Detection of spreading depolarizations in a middle cerebral artery occlusion model in swine.

The main objective of this study was to generate a hemodynamically stable swine model to detect spreading depolarizations (SDs) using electrocorticography (ECoG) and intrinsic optical signal (IOS) imaging and laser speckle flowmetry (LSF) after a 30-h middle cerebral artery (MCA) occlusion (MCAo) in German Landrace Swine. A total of 21 swine were used. The study comprised a training group (group 1, n = 7), a group that underwent bilateral craniectomy and MCAo (group 2, n = 10) and a group used for 2,3,5-triphenyltetrazolium (TTC) staining (group 3, n = 5). In group 2, nine animals that underwent MCAo survived for 30 h, and one animal survived for 12 h. We detected MCA variants with 2 to 4 vessels. In all cases, all of the MCAs were occluded. The intensity changes exhibited by IOS and LSF after clipping were closely correlated and indicated a lower blood volume and reduced blood flow in the middle cerebral artery territory. Using IOS, we detected a mean of 2.37 ± (STD) 2.35 SDs/h. Using ECoG, we detected a mean of 0.29 ± (STD) 0.53 SDs/h. Infarctions were diagnosed using histological analysis. TTC staining in group 3 confirmed that the MCA territory was compromised and that the anterior and posterior cerebral arteries were preserved. We confirm the reliability of performing live monitoring of cerebral infarctions using our MCAo protocol to detect SDs.

Measurement and visualization of stimulus-evoked tissue dynamics in mouse barrel cortex using phase-sensitive optical coherence tomography.

We describe a method to measure tissue dynamics in mouse barrel cortex during functional activation via phase-sensitive optical coherence tomography (PhS-OCT). The method measures the phase changes in OCT signals, which are induced by the tissue volume change, upon which to localize the activated tissue region. Phase unwrapping, compensation and normalization are applied to increase the dynamic range of the OCT phase detection. To guide the OCT scanning, intrinsic optical signal imaging (IOSI) system equipped with a green light laser source (532 nm) is integrated with the PhS-OCT system to provide a full field time-lapsed images of the reflectance that is used to identify the transversal 2D localized tissue response in the mouse brain. The OCT results show a localized decrease in the OCT phase signal in the activated region of the mouse brain tissue. The decrease in the phase signal may be originated from the brain tissue compression caused by the vasodilatation in the activated region. The activated region revealed in the cross-sectional OCT image is consistent with that identified by the IOSI imaging, indicating the phase change in the OCT signals may associate with the changes in the corresponding hemodynamics. In vivo localized tissue dynamics in the barrel cortex at depth during whisker stimulation is observed and monitored in this study.

Inside Front Cover: Neurosurgical brain tumor detection based on intraoperative optical intrinsic signal imaging technique: A case report of glioblastoma (J. Biophotonics 1/2020)

Intraoperative neurosurgical diagnosis of brain tumor determines the success rate of patients' prognoses. We firstly proposed a novel approach based on an optical method to distinguish the site of the tumor functionally. The promoted technique is non‐invasive, non‐radioactive, dye‐free, and potential of real‐time monitoring, which is still not available nowadays by using other techniques. The method could be applied to neuroimage guiding system for precision surgery of brain.Further details can be found in the article by Xin‐Rui Liu, Tien‐Yu Hsiao, Yun‐Qian, et al. (e201900200). image

Aging Impairs Cerebrovascular Reactivity at Preserved Resting Cerebral Arteriolar Tone and Vascular Density in the Laboratory Rat.

The age-related (mal)adaptive modifications of the cerebral microvascular system have been implicated in cognitive impairment and worse outcomes after ischemic stroke. The magnitude of the hyperemic response to spreading depolarization (SD), a recognized principle of ischemic lesion development has also been found to be reduced by aging. Here, we set out to investigate whether the SD-coupled reactivity of the pial arterioles is subject to aging, and whether concomitant vascular rarefaction may contribute to the age-related insufficiency of the cerebral blood flow (CBF) response. CBF was assessed with laser-speckle contrast analysis (LASCA), and the tone adjustment of pial arterioles was followed with intrinsic optical signal (IOS) imaging at green light illumination through a closed cranial window created over the parietal cortex of isoflurane-anesthetized young (2 months old) and old (18 months old) male Sprague–Dawley rats. Global forebrain ischemia and later reperfusion were induced by the bilateral occlusion and later release of both common carotid arteries. SDs were elicited repeatedly with topical 1M KCl. Pial vascular density was measured in green IOS images of the brain surface, while the density and resting diameter of the cortical penetrating vasculature was estimated with micro-computed tomography of paraformaldehyde-fixed cortical samples. Whilst pial arteriolar dilation in response to SD or ischemia induction were found reduced in the old rat brain, the density and resting diameter of pial cortical vessels, and the degree of SD-related oligemia emerged as variables unaffected by age in our experiments. Spatial flow distribution analysis identified an age-related shift to a greater representation of higher flow ranges in the reperfused cortex. According to our data, impairment of functional arteriolar dilation, at preserved vascular density and resting vascular tone, may be implicated in the age-related deficit of the CBF response to SD, and possibly in the reduced efficacy of neurovascular coupling in the aging brain. SD has been recognized as a potent pathophysiological contributor to ischemic lesion expansion, in part because of the insufficiency of the associated CBF response. Therefore, the age-related impairment of cerebral vasoreactivity as shown here is suggested to contribute to the age-related acceleration of ischemic lesion development.

Low-cost embedded system for optical imaging of intrinsic signals

This paper describes the proof-of-concept evaluation of a low-cost imaging system for obtaining functional connectivity maps of in vivo murine models. This non-contact system is based on the Raspberry Pi 3 and its V2 camera and offers a method for obtaining resting-state images of brain activity without the use of extrinsic contrast agents. The system was fully characterized in terms of dark signal, linearity, sensor noise resolution and spatial frequency response. One mouse was observed in vivo and functional connectivity maps were obtained by combining resting-state analysis and optical intrinsic signals imaging. Intra-mouse variations in functional connectivity remain consistent across multiple imaging sessions. In principle, inexpensive optical imaging of intrinsic signals allows the study of the mechanisms underlying human brain disorders in well-controlled murine models.

Brain segmentation, spatial censoring, and averaging techniques for optical functional connectivity imaging in mice.

Resting-state functional connectivity analysis using optical neuroimaging holds the potential to be a powerful bridge between mouse models of disease and clinical neurologic monitoring. However, analysis techniques specific to optical methods are rudimentary, and algorithms from magnetic resonance imaging are not always applicable to optics. We have developed visual processing tools to increase data quality, improve brain segmentation, and average across sessions with better field-of-view. We demonstrate improved performance using resting-state optical intrinsic signal from normal mice. The proposed methods increase the amount of usable data from neuroimaging studies, improve image fidelity, and should be translatable to human optical neuroimaging systems.

Spreading Depolarization during the Acute Stage of Experimental Subarachnoid Hemorrhage in Mice.

Spreading depolarization (SD) has been suggested as a pathomechanism for delayed cerebral ischemia after subarachnoid hemorrhage (SAH). However, the role of SD during the acute phase of SAH is still unclear. The objective of this study was to investigate (a) the occurrence of SD with intrinsic optical signal (IOS) imaging, (b) the effect of ketamine on SD, and (c) the resulting brain edema (brain water content (BWC)) during the acute stage of experimental SAH in mice. SAH was elicited by the endovascular filament perforation method. After SAH or sham operation, ketamine or saline, 30mg/kg, was given every half hour. Changes in tissue light reflectance were recorded with IOS. BWC was measured during the acute stage. Overall, 199 SDs occurred in SAH groups and 33 SDs appeared in sham groups. These SDs displayed distinct originating and spreading patterns. Compared with saline, ketamine decreased SD spread and influenced the amplitude, duration, and speed of SD. However, the occurrence of SD was not prevented by ketamine. Moreover, ketamine did not reduce BWC after SAH. These results demonstrate that SD occurs with a high incidence during the acute stage of SAH. SDs are heterogeneous in incidence, origination, and propagation. It remains unclear whether ketamine effects on SD may be viewed as therapeutically beneficial after SAH.

The 4-Aminopyridine Model of Acute Seizures in vitro Elucidates Efficacy of New Antiepileptic Drugs.

Up to date, preclinical screening for new antiepileptic substances is performed by a combination of different in vivo models of acute seizures, for which large numbers of animals are necessary. So far, little attention has been paid to in vitro models, which are also able to detect antiepileptic efficacy and in principle could likewise serve for exploratory preclinical screening. One of the established in vitro models of acute seizures is the 4-aminopyridine (4-AP) model. Previous studies have shown that the 4-AP model is capable to recapitulate the antiepileptic efficacy of standard antiepileptic drugs (AEDs) such as valproate or carbamazepine. Here, we employed a dual methodological approach using electrophysiology and optical imaging to systematically test the antiepileptic efficacy of three new-generation AEDs with distinct mechanisms of action (lacosamide, zonisamide, and levetiracetam). We found that frequency of 4-AP induced seizure like events (SLE) was the most sensitive parameter to detect dose-dependent antiepileptic effects in these compounds. Specifically, levetiracetam reduced SLE frequency while lacosamide and zonisamide at higher doses completely blocked SLE incidence. Analysis of the intrinsic optical signal additionally revealed a subiculum-specific reduction of the area involved in the propagation of ictal activity when lacosamide or zonisamide were administered. Taken together, our data adds some evidence that acute seizure models in vitro are in principle capable to detect antiepileptic effects across different mechanisms of action with efficacy similar to acute models in vivo. Further studies with negative controls, e.g., penicillin as a proconvulsant, and other clinically relevant AEDs are needed to determine if this acute in vitro model might be useful as exploratory screening tool. In view of the increasing sensitivity toward animal welfare, an affective in vitro model may help to reduce the number of laboratory animals deployed in burdening in vivo experiments and to preselect substances for subsequent testing in time- and cost-laborious models of chronic epilepsy.

Differential effects of anesthetics on resting state functional connectivity in the mouse

Blood oxygen level-dependent (BOLD) functional MRI (fMRI) is a standard approach to examine resting state functional connectivity (RSFC), but fMRI in animal models is challenging. Recently, functional optical intrinsic signal imaging—which relies on the same hemodynamic signal underlying BOLD fMRI—has been developed as a complementary approach to assess RSFC in mice. Since it is difficult to ensure that an animal is in a truly resting state while awake, RSFC measurements under anesthesia remain an important approach. Therefore, we systematically examined measures of RSFC using non-invasive, widefield optical intrinsic signal imaging under five different anesthetics in male C57BL/6J mice. We find excellent seed-based, global, and interhemispheric connectivity using tribromoethanol (Avertin) and ketamine–xylazine, comparable to results in the literature including awake animals. Urethane anesthesia yielded intermediate results, while chloral hydrate and isoflurane were both associated with poor RSFC. Furthermore, we found a correspondence between the strength of RSFC and the power of low-frequency hemodynamic fluctuations. In conclusion, Avertin and ketamine–xylazine provide robust and reproducible measures of RSFC in mice, whereas chloral hydrate and isoflurane do not.

Optical intrinsic signal imaging with optogenetics reveals functional cortico-cortical connectivity at the columnar level in living macaques

Despite extensive research on primate cognitive function, understanding how anatomical connectivity at a neural circuit level relates to information transformation across different cortical areas remains primitive. New technology is needed to visualize inter-areal anatomical connectivity in living monkeys and to tie this directly to neurophysiological function. Here, we developed a novel method to investigate this structure-function relationship, by combining optical intrinsic signal imaging (OISI) with optogenetic stimulation in living monkeys (opto-OISI). The method involves expressing channelrhodophsin-2 in one area (source) followed by optical imaging of optogenetic activations in the other area (target). We successfully demonstrated the potential of the method with interhemispheric columnar projection patterns between V1/V2 border regions. Unlike the combination of optogenetics and functional magnetic resonance imaging (opto-fMRI), opto-OISI has the advantage of enabling us to detect responses of small clusters of neurons, even if the clusters are sparsely distributed. We suggest that opto-OISI can be a powerful approach to understanding cognitive function at the neural circuit level, directly linking inter-areal circuitry to fine-scale structure and function.

Depth-Resolved Physiological Response of Retina to Transcorneal Electrical Stimulation Measured With Optical Coherence Tomography.

Transcorneal electrical stimulation (TES) has become an effective strategy to modulate retinal neural activities and partially restore visual function in ophthalmic diseases. However, the exact responses in different retinal layers still need to be clarified. This paper's goal was to evaluate the depth-resolved retinal physiological responses evoked by TES by using optical coherence tomography (OCT). A custom-built spectral-domain OCT system was used to record the intrinsic optical signals (IOSs) in different retinal layers. TES and flickers were used to stimulate the retina electrically and visually. Tetrodotoxin was used to inhibit the retinal neural activity for confirming the origin of TES-induced IOSs. We found both positive and negative IOSs could be evoked by TES in three segmented retinal layers, especially in the inner retina and subretinal space. The TES-induced IOSs correlated with the TES intensity. After tetrodotoxin injection, the IOSs evoked by TES were significantly declined, peculiarly in the inner retina. The IOSs elicited by flickers kept increasing during the stimulation, while those evoked by TES kept at a stable level. In conclusion, TES could elicit IOSs that originated from retinal neural activity in all segmented layers. The TES-induced IOSs were highly synchronized to the electrical field in the retina.

Cortical spreading depolarizations induced by surgical field blood in a mouse model of neurosurgery.

Cortical spreading depolarization (CSD) has been linked to poor clinical outcomes in the setting of traumatic brain injury, malignant stroke, and subarachnoid hemorrhage. There is evidence that electrocautery during neurosurgical procedures can also evoke CSD waves in the brain. It is unknown whether blood contacting the cortical surface during surgical bleeding affects the frequency of spontaneous or surgery-induced CSDs. Using a mouse neurosurgical model, the authors tested the hypothesis that electrocautery can induce CSD waves and that surgical field blood (SFB) is associated with more CSDs. The authors also investigated whether CSD can be reliably observed by monitoring the fluorescence of GCaMP6f expressed in neurons. CSD waves were monitored by using confocal microscopy to detect fluorescence increases at the cortical surface in mice expressing GCaMP6f in CamKII-positive neurons. The cortical surface was electrocauterized through an adjacent burr hole. SFB was simulated by applying a drop of tail vein blood to the brain through the same burr hole. CSD waves were readily detected in GCaMP6f-expressing mice. Monitoring GCaMP6f fluorescence provided far better sensitivity and spatial resolution than detecting CSD events by observing changes in the intrinsic optical signal (IOS). Forty-nine percent of the CSD waves identified by GCaMP6f had no corresponding IOS signal. Electrocautery evoked CSD waves. On average, 0.67 ± 0.08 CSD events were generated per electrocautery episode, and multiple CSD waves could be induced in the same mouse by repeated cauterization (average, 7.9 ± 1.3 events; maximum number in 1 animal, 13 events). In the presence of SFB, significantly more spontaneous CSDs were generated (1.35 ± 0.37 vs 0.13 ± 0.16 events per hour, p = 0.002). Ketamine effectively decreased the frequency of spontaneous CSD waves (1.35 ± 0.37 to 0.36 ± 0.15 CSD waves per hour, p = 0.016) and electrocautery-stimulated CSD waves (0.80 ± 0.05 to 0.18 ± 0.08 CSD waves per electrocautery, p = 0.00002). CSD waves are detected with far greater sensitivity and fidelity by monitoring GCaMP6f signals in neurons than by monitoring IOSs. Electrocautery reliably evokes CSD waves, and the frequency of spontaneous CSD waves is increased when blood is applied to the cortical surface. These experimental conditions recapitulate common scenarios in the neurosurgical operating room. Ketamine, a clinically available pharmaceutical agent, can block stimulated and spontaneous CSDs. More research is required to understand the clinical importance of intraoperative CSD.

Causal relationship between local field potential and intrinsic optical signal in epileptiform activity in vitro

The directed causal relationship were examined between the local field potential (LFP) and the intrinsic optical signal (IOS) during induced epileptiform activity in in vitro cortical slices by the convergent cross-mapping causality analysis method. Two components of the IOS signal have been distinguished: a faster, activity dependent component (IOSh) which changes its sign between transmitted and reflected measurement, thus it is related to the reflectance or the scattering of the tissue and a slower component (IOSl), which is negative in both cases, thus it is resulted by the increase of the absorption of the tissue. We have found a strong, unidirectional, delayed causal effect from LFP to IOSh with 0.5-1s delay, without signs of feedback from the IOSh to the LFP, while the correlation was small and the peaks of the cross correlation function did not reflect the actual causal dependency. Based on these observations, a model has been set up to describe the dependency of the IOSh on the LFP power and IOSh was reconstructed, based on the LFP signal. This study demonstrates that causality analysis can lead to better understanding the physiological interactions, even in case of two data series with drastically different time scales.

Pharmacological Dissection of Intrinsic Optical Signal Reveals a Functional Coupling between Synaptic Activity and Astrocytic Volume Transient.

The neuronal activity-dependent change in the manner in which light is absorbed or scattered in brain tissue is called the intrinsic optical signal (IOS), and provides label-free, minimally invasive, and high spatial (~100 µm) resolution imaging for visualizing neuronal activity patterns. IOS imaging in isolated brain slices measured at an infrared wavelength (>700 nm) has recently been attributed to the changes in light scattering and transmittance due to aquaporin-4 (AQP4)-dependent astrocytic swelling. The complexity of functional interactions between neurons and astrocytes, however, has prevented the elucidation of the series of molecular mechanisms leading to the generation of IOS. Here, we pharmacologically dissected the IOS in the acutely prepared brain slices of the stratum radiatum of the hippocampus, induced by 1 s/20 Hz electrical stimulation of Schaffer-collateral pathway with simultaneous measurement of the activity of the neuronal population by field potential recordings. We found that 55% of IOSs peak upon stimulation and originate from postsynaptic AMPA and NMDA receptors. The remaining originated from presynaptic action potentials and vesicle fusion. Mechanistically, the elevated extracellular glutamate and K+ during synaptic transmission were taken up by astrocytes via a glutamate transporter and quinine-sensitive K2P channel, followed by an influx of water via AQP-4. We also found that the decay of IOS is mediated by the DCPIB- and NPPB-sensitive anion channels in astrocytes. Altogether, our results demonstrate that the functional coupling between synaptic activity and astrocytic transient volume change during excitatory synaptic transmission is the major source of IOS.

Near infrared imaging of intrinsic signals in cortical spreading depression observed through the intact scalp in hairless mice

Brain cooling was inevitable in both thinned and intact skull windows in neuroimaging in vivo of mice. Thus we proposed the novel imaging method leaving intact scalp on the skull using the light at 670, 785, and 975 nm. In this study, we used hairless mice (Hos:HR-1) since the deterioration of image quality was resulted from the hair. Cortical spreading depression was induced by KCl application through small incision and burr hole on the frontal bone. Intrinsic optical signals through the intact scalp in the observation area were detected. Time course of the signal showed a triphasic feature which was consistent with the intrinsic optical signals through the intact skull. In three pairs of signal amplitudes at the different wavelengths, no significant differences were observed. Although the intact scalp weakened the amplitudes significantly, e.g., 4.0 from 6.9 at 975 nm, the signals during cortical spreading depression were sufficient to be detected.

Abstract P6-09-12: Using multiphoton laser scanning microscopy to assess neoadjuvant therapy outcome in core needle biopsies: A novel methodology

Abstract Background: Over-expression of Human Epidermal Growth Factor receptor-2 (HER2) in breast cancer is associated with an aggressive clinical course and poor prognosis. Targeting HER2 over-expression has been shown to be a remarkably effective therapeutic modality in the metastatic, adjuvant and neoadjuvant setting and the pathologic response to neoadjuvant treatment in HER2-positive breast cancer has been shown to be an excellent surrogate for a good outcome. The stromal tumor microenvironment is implicated in fostering tumor growth, facilitating cell migration and ultimately resulting in metastatic disease. Specifically, the collagenous extracellular matrix, which includes fibrillar collagen, has been suggested to play a role in the migration of malignant breast epithelial cells within the surrounding stroma. We have developed a novel methodology which uses an intrinsic optical signature to quantitatively evaluate fibrillar collagen (Burke et al. BMC Cancer 15 (2015): 929). Here, we evaluate the ability of this quantitative methodology to predict the pathologic response after neoadjuvant HER2-targeted treatment as assessed by the Residual Cancer Burden score/class (RCB). This quantitative evaluation in pre-treatment biopsies is then correlated with the pathologic response to treatment in the post-therapy resection. Material and Methods: Clinical pathologic variables for 29 cases of HER2-positive breast cancer that had undergone neoadjuvant chemotherapy plus HER2-targeted therapy were retrieved from the medical record database at URMC, including the post-treatment RCB score and ER/PR/HER2 status. Second harmonic generation (SHG) is an intrinsic optical signal produced by fibrillar collagen. To quantify collagen microstructure in the pre-treatment core biopsy, we used SHG imaging to determine the average forward to backward-light scattering ratio (F/B). The F/B ratio is sensitive to structural properties of collagen fibers. Results: Logistic regression was used to assess the association between F/B and the binary response variable RCB class (0/1 or 2/3). A likelihood ratio test was used to calculate the p-value to test whether the regression coefficient for F/B was zero (i.e. no effect) in the tumor-stromal interface. The average F/B ratio at the leading edge of the tumor stratified by RCB class is shown in Table 1. When evaluated in the bulk of the tumor tissue, F/B was not correlated with RCB status; however, when evaluated at the leading edge of the tumor stromal interface, F/B was significantly correlated with RCB status (p=0.035). Table 1:RCB class and average F/BRCB class (n)Average F/B ± SEM0/1 (19)16.95 ± 1.062/3 (10)12.32 ± 1.84 Conclusions: We have previously shown that the measurement of F/B in the primary tumor after resection is an independent prognostic indicator of metastasis-free survival in breast cancer. Our results in the current study furthers these observations and suggests that the evaluations of the microstructure of collagen fibers by F/B measurement from the pre-treatment biopsy, specifically at the leading edge of the tumor-stroma interface, may be useful for predicting pathologic response to trastuzumab-based neoadjuvant therapy. Further studies in a larger patient cohort are warranted. Citation Format: Desa DE, Turner BM, Buscaglia B, Hill RL, Strawderman RL, Hicks DG, Brown EB. Using multiphoton laser scanning microscopy to assess neoadjuvant therapy outcome in core needle biopsies: A novel methodology [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P6-09-12.

In vivo imaging for neurovascular disease research

Connections between various cell types in the brain enable cognitive function. The neurovascular unit is a structure composed of different cell types that regulate neurovascular coupling, blood-brain barrier permeability, and other interactions with peripheral systems. The relationship among the components of the neurovascular unit is complex and difficult to study without the use of in vivo neurovascular disease imaging. In this review, we introduce principles and examples of various in vivo optical imaging techniques including laser Doppler flowmetry, laser speckle contrast imaging, intrinsic optical signal imaging, optical coherence tomography, and two-photon microscopy. Furthermore, we introduce recent advances of in vivo imaging and future directions for promoting neurovascular disease research.

Local Changes in Water Balance as a Marker of Neuron Activity in the Somatosensory System in Neonatal Rat Pups

Recording of intrinsic optical signals (IOS) is widely used for functional mapping in the central nervous system. However, the effectiveness of the IOS method depends critically on the chromophore used. The most commonly used chromophores for detection of IOS in living organisms are the oxygenated and deoxygenated forms of hemoglobin, which provide for indirect assessment of neuron activity in terms of changes in oxygenated blood levels. However, in some cases these changes are weak, decreasing the effectiveness of the IOS method. We report here our studies of the potential for using water as the chromophore in the spectral recording of IOS. Stimulation of the sensory whiskers on the snouts of neonatal rat pups induced IOS in the corresponding parts of the animals’ sensorimotor cortex. Evoked IOS were recorded in the near infrared (IR) part of the spectrum. Shifts in the recorded spectral range were accompanied by decreases in the spontaneous noise level which, despite the low signal amplitude, led to a significant increase in the signal: noise ratio. These results provide evidence of the potential for using water as a marker for neural activity using the IOS recording method.

More Light on the Brain: 30 Years Later

Recent decades have been characteristics by the rapid development of optical methods in neurobiology. Historically, the first optical methods of recording neuron activity were those using potential-dependent dyes with recording of the overall activity of small areas of the brain exploiting intrinsic optical signals. With the development of genetic technologies, optogenetics appeared, allowing not only neuron activity to be recorded optically but also cell operation to be controlled using photostimulation. The wide use of optogenetics in neurobiological research has filled a significant gap in approaches to studies of brain function in the last decade. Active attempts are now being made to use optogenetic approaches in the treatment of epilepsy and a series of neurodegenerative diseases in clinical practice.

Modulating brain functional connectivity using transcranial ultrasound stimulation

Transcranial ultrasound stimulation can modulate the brain non-invasively using focused acoustic waves with high resolution. In previous studies, the stimulation has been shown to increase cortical excitability and improve task performance [1], as well as induce mood changes [2] without lasting negative effects. In addition, changes were observed to persist for more than half an hour after the stimulation [3]. These outcomes show possible evidence of alterations in the brain functional connectivity induced by stimulation. In this work, we present an ultrasound stimulation system coupled with neuroimaging based on optical intrinsic signals for examining the brain functional connectivity. Optical intrinsic signal imaging (OISI) is a convenient, low-cost and less bulky alternative to study the effects of ultrasound neuromodulation in small animals compared to the conventional fMRI. Ultrasound stimulation was achieved using a custom high frequency 2 MHz ultrasound transducer with a sub-1 mm focal radius. Using the combined system, we examine how ultrasound stimulation affects the brain functional connectivity in a mouse model.