Brain metabolism changes in cases of impaired breathing or blood circulation in rodents evaluated by real time optical spectroscopy methods

The aim of the study was to compare the metabolic activity of brain cortex after the acute hypoxia caused by the impairment of breathing or blood circulation. Male Wistar rats were randomized in two groups: impaired breathing and blood circulation failure groups. Fluorescence under 365 and 450 nm excitation and diffuse reflectance intensity at 550-820 nm range were estimated. We found that after long-term hypoxic conditions, notable metabolic changes occur. We suppose that oxygen deficiency causes an activation of the GABA shunt mechanism. In cases of blood circulation failure, fluorescence intensity changes faster than in cases of breathing impairment

Impairments of cerebral blood flow microcirculation in rats brought on by cardiac cessation and respiratory arrest

The impairments of cerebral blood flow microcirculation brought on by cardiac and respiratory arrest were assessed with multi-modal diagnostic facilities, utilising laser speckle contrast imaging, fluorescence spectroscopy and diffuse reflectance spectroscopy. The results of laser speckle contrast imaging show a notable reduction of cerebral blood flow in small and medium size vessels during a few minutes of respiratory arrest, while the same effect was observed in large sinuses and their branches during the circulatory cessation. Concurrently, the redox ratio assessed with fluorescence spectroscopy indicates progressing hypoxia, NADH accumulation and increase of FAD consumption. The results of diffuse reflectance spectra measurements display a more rapid grow of the perfusion of deoxygenated blood in case of circulatory impairment. In addition, consequent histopathological analysis performed by using new tissue staining procedure developed in-house. It shows notably higher reduction of size of the neurons due to their wrinkling within brain tissues influenced by circulation impair. Whereas, the brain tissues altered with the respiratory arrest demonstrate focal perivascular oedema and mild hypoxic changes of neuronal morphology. Thus, the study suggests that consequences of a cessation of cerebral blood flow become more dramatic and dangerous compare to respiratory arrest

Optical imaging and spectroscopy for the study of the human brain: status report

This report is the second part of a comprehensive two-part series aimed at reviewing an extensive and diverse toolkit of novel methods to explore brain health and function. While the first report focused on neurophotonic tools mostly applicable to animal studies, here, we highlight optical spectroscopy and imaging methods relevant to noninvasive human brain studies. We outline current state-of-the-art technologies and software advances, explore the most recent impact of these technologies on neuroscience and clinical applications, identify the areas where innovation is needed, and provide an outlook for the future directions

Optical imaging and spectroscopy for the study of the human brain: status report.

This report is the second part of a comprehensive two-part series aimed at reviewing an extensive and diverse toolkit of novel methods to explore brain health and function. While the first report focused on neurophotonic tools mostly applicable to animal studies, here, we highlight optical spectroscopy and imaging methods relevant to noninvasive human brain studies. We outline current state-of-the-art technologies and software advances, explore the most recent impact of these technologies on neuroscience and clinical applications, identify the areas where innovation is needed, and provide an outlook for the future directions

Optical imaging and spectroscopy for the study of the human brain: status report

This report is the second part of a comprehensive two-part series aimed at reviewing an extensive and diverse toolkit of novel methods to explore brain health and function. While the first report focused on neurophotonic tools mostly applicable to animal studies, here, we highlight optical spectroscopy and imaging methods relevant to noninvasive human brain studies. We outline current state-of-the-art technologies and software advances, explore the most recent impact of these technologies on neuroscience and clinical applications, identify the areas where innovation is needed, and provide an outlook for the future directions. Keywords: DCS; NIRS; diffuse optics; functional neuroscience; optical imaging; optical spectroscop

Advances in Dynamic Light Scattering Imaging of Blood Flow

Dynamic light scattering (DLS) is a well known experimental approach uniquely suited for the characterization of small particles undergoing Brownian motion in randomly inhomogeneous turbid scattering medium, including water suspension, polymers in solutions, cells cultures, and so on. DLS is based on the illuminating of turbid medium with a coherent laser light and further analyzes the intensity fluctuations caused by the motion of the scattering particles. The DLS-based spin-off derivative techniques, such laser Doppler flowmetry (LDF), diffusing wave spectroscopy (DWS), laser speckle contrast imaging (LSCI), and Doppler optical coherence tomography (DOCT), are exploited widely for non-invasive imaging of blood flow in brain, skin, muscles, and other biological tissues. The recent advancements in the DLS-based imaging technologies in frame of their application for brain blood flow monitoring, skin perfusion measurements, and non-invasive blood micro-circulation characterization are overviewed. The fundamentals, breakthrough potential, and practical findings revealed by DLS-based blood flow imaging studies, including the limitations and challenges of the approach such as movement artifacts, non-ergodicity, and overcoming high scattering properties of studied medium, are also discussed. It is concluded that continued research and further technological advancements in DLS-based imaging will pave the way for new exciting developments and insights into blood flow diagnostic imaging

Quantitative cerebral blood flow with optical coherence tomography

Absolute measurements of cerebral blood flow (CBF) are an important endpoint in studies of cerebral pathophysiology. Currently no accepted method exists for in vivo longitudinal monitoring of CBF with high resolution in rats and mice. Using three-dimensional Doppler Optical Coherence Tomography and cranial window preparations, we present methods and algorithms for regional CBF measurements in the rat cortex. Towards this end, we develop and validate a quantitative statistical model to describe the effect of static tissue on velocity sensitivity. This model is used to design scanning protocols and algorithms for sensitive 3D flow measurements and angiography of the cortex. We also introduce a method of absolute flow calculation that does not require explicit knowledge of vessel angles. We show that OCT estimates of absolute CBF values in rats agree with prior measures by autoradiography, suggesting that Doppler OCT can perform absolute flow measurements in animal models.National Institutes of Health (U.S.) (Grant number R01-NS057476)National Institutes of Health (U.S.) (Grant number P01NS055104)National Institutes of Health (U.S.) (Grant number P50NS010828)ational Institutes of Health (U.S.) (Grant number K99NS067050)National Institutes of Health (U.S.) (Grant number R01-CA075289-13)United States. Air Force Office of Scientific Research (FA9550-07-1-0014)United States. Dept. of Defense. Medical Free Electron Laser Program (FA9550-07-1-0101

Time resolved speckle contrast optical spectroscopy at quasi-null source-detector separation for non-invasive measurement of microvascular blood flow

Time (or path length) resolved speckle contrast optical spectroscopy (TD-SCOS) at quasi-null (2.85 mm) source-detector separation was developed and demonstrated. The method was illustrated by in vivo studies on the forearm muscle of an adult subject. The results have shown that selecting longer photon path lengths results in higher hyperemic blood flow change and a faster return to baseline by a factor of two after arterial cuff occlusion when compared to SCOS without time resolution. This indicates higher sensitivity to the deeper muscle tissue. In the long run, this approach may allow the use of simpler and cheaper detector arrays compared to time resolved diffuse correlation spectroscopy that are based on readily available technologies. Hence, TD-SCOS may increase the performance and decrease cost of devices for continuous non-invasive, deep tissue blood flow monitoring

Development of an Awake Behaving model for Laser Doppler Flowmetry in Mice

Bien que le cerveau ne constitue que 2% de la masse du corps chez les humains, il présente l’activité métabolique la plus élevée dans le corps, et en conséquence, constitue un organe hautement vascularisé. En fait, l’approvisionnement en sang dans le cerveau est strictement modulé au niveau régional par un mécanisme fondamental nommé couplage neurovasculaire (CNV), qui associe les besoins métaboliques locaux au flux sanguin cérébral [1, 2]. Notre compréhension du CNV sous des conditions physiologiques et pathologiques a été améliorée par un large éventail d’études menées chez les rongeurs. Néanmoins, ces études ont été réalisées soit sous anesthésie, soit chez la souris éveillée et immobilisée, afin d’éviter le mouvement de la tête pendant l'acquisition de l'image. Les anesthésiques, ainsi que le stress induit par la contention, peuvent altérer l'hémodynamique cérébrale, ce qui pourrait entraver les résultats obtenus. Par conséquent, il est essentiel de contrôler ces facteurs lors de recherches futures menées au sujet de la réponse neurovasculaire. Au cours de l’étude présente, nous avons développé un nouveau dispositif pour l'imagerie optique éveillée, où la tête de la souris est immobilisée, mais son corps est libre de marcher, courir ou se reposer sur une roue inclinée. En outre, nous avons testé plusieurs protocoles d'habituation, selon lesquels la souris a été progressivement entraînée pour tolérer l’immobilisation de tête, afin de minimiser le stress ressenti lors des sessions d'imagerie. Enfin, nous avons, pour la première fois, cherché à valider l'efficacité de ces protocoles d'habituation dans la réduction du stress, en mesurant l'évolution des taux plasmatiques de corticostérone tout au long de notre étude. Nous avons noté que les souris s'étaient rapidement adaptées à la course sur la roue et que les signes visibles de stress (luttes, vocalisations et urination) étaient nettement réduits suite à deux sessions d'habituation. Néanmoins, les taux de corticostérone n'ont pas été significativement réduits chez les souris habituées, par rapport aux souris naïves qui ont été retenues sur la roue sans entraînement préalable (p> 0,05). Ce projet met en évidence la nécessité d'une mesure quantitative du stress, car une réduction des comportements observables tels que l'agitation ou la lutte peut être indicative non pas d'un niveau de stress plus faible, mais plutôt d'un désespoir comportemental. Des recherches supplémentaires sont nécessaires pour déterminer si la fixation de la tête lors de l'imagerie optique chez la souris peut être obtenue avec des niveaux de stress plus faibles, et si le stress induit par la contrainte effectuée avec notre dispositif est associé à des changements de la réponse hémodynamique.Whilst the brain only constitutes 2% of total body weight in humans, it exhibits the highest metabolic activity in the body, and as such is a highly vascularized organ. In fact, regional blood supply within the brain is strictly modulated through a fundamental process termed neurovascular coupling (NVC), which couples local metabolic needs with cerebral blood flow [1, 2]. A wide array of optical imaging studies in rodents has enhanced our understanding of NVC under physiological and pathological conditions. Nevertheless, these studies have been performed either under anesthesia, or in the awake mouse using restraint to prevent head-motion during image acquisition. Both anesthetics and restraint-induced stress have been clearly shown to alter cerebral hemodynamics, thereby potentially interfering with the obtained results [3, 4]. Hence, it is essential to control for these factors during future research which investigates the neurovascular response. In the present study, we have developed a new apparatus for awake optical imaging, where the mouse is head-restraint whilst allowed to walk, run or rest on an inclined wheel. In addition, we have tested several habituation protocols, according to which the mouse was gradually trained to tolerate head-restraint, in order to minimize the stress experienced during imaging sessions. Lastly, we have, for the first time, sought to validate the efficiency of these habituation protocols in reducing stress, by measuring the evolution of plasma corticosterone levels throughout the study. We noted that the mice had quickly adapted to running on the wheel, and that the overt signs of stress (struggling, vocalizations and urination) were clearly reduced within two habituation sessions. Nevertheless, corticosterone levels were not significantly reduced in habituated mice, relative to naïve mice that were restrained on the wheel without prior training (p > 0.05). This project highlights the necessity for a quantitative measure of stress, as a reduction in observable behaviors such as agitation or struggling may be indicative not of lower stress, but rather, of behavioral despair. Further research is needed to determine whether head-fixation during optical imaging in mice can be achieved with lower stress levels, and if restraint-induced stress using our apparatus is associated with changes in the hemodynamic response

EVALUATION OF ELECTROPHYSIOLOGICAL PROGNOSTIC TOOLS TO TRACK BRAIN RECOVERY WITH TEMPERATURE MANAGEMENT AFTER CARDIAC ARREST

Cardiac arrest (CA) occurs in over 347,000 adult Americans annually and is a leading cause of morbidity and mortality. While targeted temperature management (TTM), a neuroprotective treatment, is often implemented following resuscitation, CA typically results in an ischemic brain injury that is detrimental to functional outcome. Prognosis shortly following CA is crucial as it may guide subsequent treatment. Although multiple prognostic tools exist, many have not been verified under TTM or have limitations in early prognostication. Thus, a reliable tool that predicts functional outcome during the early recovery period after CA is required. For this project, it was necessary to first review current literature detailing the existing prognostic tools and their limitations. Many of the prognostic tools that have been employed have limitations including subjective results interpretation and the confounding effects of the sedatives required for TTM. However, somatosensory evoked potentials (SSEP) and electroencephalogram (EEG) have potential to track recovery. The bilateral absence of the human N20 SSEP peak is currently the most reliable predictor of poor outcome, however, the signal interpretation is subjective and limited by the dichotomous categorization. Thus, the next step in this work included quantitative analyses of SSEP signals to identify objective prognostic markers. First, the peak amplitude and latency of SSEP signals were calculated. The amplitude of N10 peaks and latency of N7 and N10 peaks in rats were measured objectively and were distinct among temperature and outcome groups. A more complex and novel calculation of the SSEP phase space area (qSSEP-PSA), which considers multiple SSEP characteristics, was then performed. Animals treated with hypothermia tended to have higher qSSEP-PSA values and better qSSEP-PSA recovery over the early recovery period. These analyses demonstrate that quantitative SSEPs can track brain recovery shortly after resuscitation. EEG is a common brain-monitoring tool. The information quantity (IQ), a quantitative measure of the EEG information content, was calculated in post-CA rats that underwent laser speckle contrast imaging (LSCI), which measures relative cerebral blood flow (rCBF). IQ was significantly and negatively correlated with rCBF during the early recovery, suggesting that electrical activity recovery can be maintained by lower rCBF shortly after CA