A novel GCaMP6f-RCS rat model for studying electrical stimulation in the degenerated retina.

Background: Retinal prostheses aim to restore vision by electrically stimulating the remaining viable retinal cells in Retinal Degeneration (RD) cases. Research in this field necessitates a comprehensive analysis of retinal ganglion cells' (RGCs) responses to assess the obtained visual acuity and quality. Here we present a novel animal model which facilitates the optical recording of RGCs activity in an RD rat. This model can significantly enhance the functional evaluation of vision restoration treatments. Methods: The development of the novel rat model is based on crossbreeding a retinal degenerated Royal College of Surgeons (RCS) rat with a transgenic line expressing the genetic calcium indicator GCaMP6f in the RGCs. Characterization of the model was achieved using Optical Coherence Tomography (OCT) imaging, histology, and electroretinography (ERG) at the ages of 4, 8, and 12 weeks. Additionally, optical recordings of RGCs function in response to ex-vivo subretinal electrical stimulations were performed. Results: Histological investigations confirmed the high expression of GCaMP6f in the RGCs and minimal expression in the inner nuclear layer (INL). OCT imaging and histological studies revealed the expected gradual retinal degeneration, as evident by the decrease in retinal thickness with age and the formation of subretinal debris. This degeneration was further confirmed by ERG recordings, which demonstrated a significant decrease in the b-wave amplitude throughout the degeneration process, culminating in its absence at 12 weeks in the GCaMP6f-RCS rat. Importantly, the feasibility of investigating subretinal stimulation was demonstrated, revealing a consistent increase in activation threshold throughout degeneration. Furthermore, an increase in the diameter of the activated area with increasing currents was observed. The spatial spread of the activation area in the GCaMP6f-RCS rat was found to be smaller and exhibited faster activation dynamics compared with the GCaMP6f-LE strain. Conclusion: This novel animal model offers an opportunity to deepen our understanding of prosthetically induced retinal responses, potentially leading to significant advancements in prosthetic interventions in visual impairments.

Optimizing the fabrication of a 3D high-resolution implant for neural stimulation.

BACKGROUND: Tissue-integrated micro-electronic devices for neural stimulation hold great potential in restoring the functionality of degenerated organs, specifically, retinal prostheses, which are aimed at vision restoration. The fabrication process of 3D polymer-metal devices with high resolution and a high aspect-ratio (AR) is very complex and faces many challenges that impair its functionality. APPROACH: Here we describe the optimization of the fabrication process of a bio-functionalized 3D high-resolution 1mm circular subretinal implant composed of SU-8 polymer integrated with dense gold microelectrodes (23µm pitch) passivated with 3D micro-well-like structures (20µm diameter, 3µm resolution). The main challenges were overcome by step-by-step planning and optimization while utilizing a two-step bi-layer lift-off process; bio-functionalization was carried out by N2 plasma treatment and the addition of a bio-adhesion molecule. MAIN RESULTS: In-vitro and in-vivo investigations, including SEM and FIB cross section examinations, revealed a good structural design, as well as a good long-term integration of the device in the rat sub-retinal space and cell migration into the wells. Moreover, the feasibility of subretinal neural stimulation using the fabricated device was demonstrated in-vitro by electrical activation of rat's retina. CONCLUSIONS: The reported process and optimization steps described here in detail can aid in designing and fabricating retinal prosthetic devices or similar neural implants.

Perceptual learning based on a temporal stimulus enhances visual function in adult amblyopic subjects.

Studies have shown that Perceptual Learning (PL) can lead to enhancement of spatial visual functions in amblyopic subjects. Here we aimed to determine whether a simple flickering stimulus can be utilized in PL to enhance temporal function performance and whether enhancement will transfer to spatial functions in amblyopic subjects. Six adult amblyopic and six normally sighted subjects underwent an evaluation of their performance of baseline psychophysics spatial functions (Visual acuity (VA), contrast sensitivity (CS), temporal functions (critical fusion frequency (CFF) test), as well as a static and flickering stereopsis test, and an electrophysiological evaluation (VEP). The subjects then underwent 5 training sessions (on average, a total of 150 min over 2.5 weeks), which included a task similar to the CFF test using the method of constant stimuli. After completing the training sessions, subjects repeated the initial performance evaluation tasks. All amblyopic subjects showed improved temporal visual performance (CFF) in the amblyopic eye (on average, 17%, p << 0.01) following temporal PL. Generalization to spatial, spatio-temporal, and binocular tasks was also found: VA increased by 0.12 logMAR (p = 0.004), CS in backward masking significantly increased (by up to 19%, p = 0.003), and flickering stereopsis increased by 85 arcsec (p = 0.048). These results were further electrophysiologically manifested by an increase in VEP amplitude (by 43%, p = 0.03), increased Signal-to-Noise ratio (SNR) (by 39%, p = 0.024) to levels not different from normally sighted subjects, along with an improvement in inter-ocular delay (by 5.8 ms, p = 0.003). In contrast, no significant effect of training was found in the normally sighted group. These results highlight the potential of PL based on a temporal stimulus to improve the temporal and spatial visual performance in amblyopes. Future work is needed to optimize this method for clinical applications.

Temporal synchronization elicits enhancement of binocular vision functions.

Integration of information over the CNS is an important neural process that affects our ability to perceive and react to the environment. The visual system is required to continuously integrate information arriving from two different sources (the eyes) to create a coherent percept with high spatiotemporal precision. Although this neural integration of information is assumed to be critical for visual performance, it can be impaired under some pathological or developmental conditions. Here we took advantage of a unique developmental condition, amblyopia ("lazy eye"), which is characterized by an impaired temporal synchronization between the two eyes, to meticulously study the effect of synchronization on the integration of binocular visual information. We measured the eyes' asynchrony and compensated for it (with millisecond temporal resolution) by providing time-shifted stimuli to the eyes. We found that the re-synchronization of the ocular input elicited a significant improvement in visual functions, and binocular functions, such as binocular summation and stereopsis, were regained. This phenomenon was also evident in neurophysiological measures. Our results can shed light on other neural processing aspects and might also have translational relevance for the field of training, rehabilitation, and perceptual learning.

Remote photonic sensing of action potential in mammalian nerve cells via histogram-based analysis of temporal spatial acoustic vibrations.

Label free and remote action potential detection in neurons can be of great importance in the neuroscience research field. This paper presents a novel label free imaging modality based on the detection of temporal vibrations of speckle patterns illuminating the sample. We demonstrated the feasibility of detecting action potentials originating from spontaneous and stimulated activity in cortical cell culture. The spatiotemporal vibrations of isolated cortical cells were extracted by illuminating the culture with a laser beam while the vibrations of the random back scattered secondary speckle patterns are captured by a camera. The postulated action potentials were estimated following correlation-based analysis on the captured vibrations, where the variance deviation of the signal from a Gaussian distribution is directly associated with the action potential events. The technique was validated in a series of experiments in which the optical signals were acquired concurrently with microelectrode array (MEA) recordings. Our results demonstrate the ability of detecting action potential events in mammalian cells remotely via extraction of acoustic vibrations.

Evaluation and Optimization of Methods for Generating High-Resolution Retinotopic Maps Using Visual Cortex Voltage-Sensitive Dye Imaging.

The localization and measurement of neuronal activity magnitude at high spatial and temporal resolution are essential for mapping and better understanding neuronal systems and mechanisms. One such example is the generation of retinotopic maps, which correlates localized retinal stimulation with the corresponding specific visual cortex responses. Here we evaluated and compared seven different methods for extracting and localizing cortical responses from voltage-sensitive dye imaging recordings, elicited by visual stimuli projected directly on the rat retina by a customized projection system. The performance of these methods was evaluated both qualitatively and quantitatively by means of two cluster separation metrics, namely, the (adjusted) Silhouette Index (SI) and the (adjusted) Davies-Bouldin Index (DBI). These metrics were validated using simulated data, which showed that Temporally Structured Component Analysis (TSCA) outperformed all other analysis methods for localizing cortical responses and generating high-resolution retinotopic maps. The analysis methods, as well as the use of cluster separation metrics proposed here, can facilitate future research aiming to localize specific activity at high resolution in the visual cortex or other brain areas.

Cortical responses to prosthetic retinal stimulation are significantly affected by the light-adaptive state of the surrounding normal retina.

Objective.Restoration of central vision loss in patients with age-related macular degeneration (AMD) by implanting a retinal prosthesis is associated with an intriguing situation wherein the central prosthetic vision co-exists with natural normal vision. Of major interest are the interactions between the prosthetic and natural vision. Here we studied the effect of the light-adaptive state of the normal retina on the electrical visual evoked potentials (VEPs) arising from the retinal prosthesis.Approach.We recorded electrical VEP elicited by prosthetic retinal stimulation in wild-type rats implanted with a 1 mm photovoltaic subretinal array. Cortical responses were recorded following overnight dark adaption and compared to those recorded following bleaching of the retina by light (520 nm) at various intensities and durations.Main results.Compared to dark-adapted responses, bleaching induced a 2-fold decrease in the prosthetic cortical response, which returned to the dark-adapted baseline within 30 min to several hours, depending on the degree of bleaching. This reduction was neither observed in Royal College of Surgeons (RCS) rats with a degenerated photoreceptor layer nor following intravitreal injection of a GABAa receptor blocker (bicuculine), suggesting the involvement of photoreceptors and a GABAa-mediated mechanism.Significance.These findings show a robust effect of the retinal light-adaptive state on the obtained prosthetic responses. If a similar effect is found in humans, this will have immediate implications on the design of prosthetic devices, where both natural and prosthetic vision co-exist, such as in AMD patients receiving a photovoltaic retinal implant. Similarly, standardization of the retinal light-adaptive state in prosthetic clinical trials should be considered.

A dichoptic presentation device and a method for measuring binocular temporal function in the visual system.

Recent studies highlight the importance of the temporal domain in visual processing. Critical Flicker Frequency (CFF), the frequency at which a flickering light is perceived as continuous, is a widely used measure for evaluating visual temporal processing. Another important issue to investigate is the cortical interactions arising between the flicker stimuli of both eyes. This paper presents a robust and reliable dichoptic tool for evaluating the CFF threshold in both eyes. This system is based on an analog output device used to independently drive two LEDs through a custom-written MATLAB code (using a laptop PC) for eliciting sinusoidal flickering stimuli and for psychophysically measuring the perceived CFF threshold. The luminance and phases of each LED are individually controlled, enabling the investigation of the effect of phase and luminance differences on binocular summation in subjects with different ocular pathologies. Experiments were designed to evaluate the CFF threshold through a psychophysical test, based on a discrimination task with a stimulus duration of 1 s, based on a temporal alternative forced-choice paradigm. The target stimulus temporal features were modulated using the staircase method. Subjects were requested to discriminate between a target stimulus (a flickering light at various frequencies) and a flickering light at a frequency of 120 Hz, which is significantly higher than the CFF in humans; therefore, it is perceived as constant. One of the main advantages of the introduced dichoptic presentation system is that it enables the visual temporal performance to be measured under both monocular and binocular conditions where phenomena such as temporal binocular summation (BS) can be evaluated. Moreover, the system offers great flexibility by introducing a stimulus phase shift, which enables studying how stimulus timing affects the temporal function at millisecond scale resolution. Our results confirm that no crosstalk exists between the eyes and that the system can reliably separate the stimuli presented to the eyes. Using this set-up, we observed the binocular summation of CFF for low target luminance levels. The CFF was significantly (p = 0.011) higher (5.2%) under binocular compared with monocular viewing conditions. More importantly, introducing an inter - ocular phase shift reduced the binocular CFF in normally sighted subjects. Finally, in amblyopic subjects the amblyopic eye showed a decrease of 3.9 Hz (15%) in CFF, compared with the fellow eye (p = 0.001). The ability to assess binocular temporal performance using a dichoptic display can shed light on visual temporal performance in general, and on binocular temporal summation processes in particular, both for subjects with normal binocular vision and for subjects with impaired binocular vision (e.g., amblyopic subjects). Furthermore, such a presentation set-up may facilitate the development of training paradigms aimed at improving binocular vision performance. In this paper we describe the system and methods in detail and provide all necessary computer code and other details that will enable an easy and quick adaptation of the method by scientists interested in studying the temporal resolution of the visual system in general, and in studying inter-ocular differences or interactions in particular.

Electrophysiologic Characterization of Developing Human Embryonic Stem Cell-Derived Photoreceptor Precursors.

Purpose: Photoreceptor precursor cells (PRPs) differentiated from human embryonic stem cells can serve as a source for cell replacement therapy aimed at vision restoration in patients suffering from degenerative diseases of the outer retina, such as retinitis pigmentosa and AMD. In this work, we studied the electrophysiologic maturation of PRPs throughout the differentiation process. Methods: Human embryonic stem cells were differentiated into PRPs and whole-cell recordings were performed for electrophysiologic characterization at days 0, 30, 60, and 90 along with quantitative PCR analysis to characterize the expression level of various ion channels, which shape the electrophysiologic response. Finally, to characterize the electrically induced calcium currents, we employed calcium imaging (rhod4) to visualize intracellular calcium dynamics in response to electrical activation. Results: Our results revealed an early and steady presence (approximately 100% of responsive cells) of the delayed potassium rectifier current. In contrast, the percentage of cells exhibiting voltage-gated sodium currents increased with maturation (from 0% to almost 90% of responsive cells at 90 days). Moreover, calcium imaging revealed the presence of voltage-gated calcium currents, which play a major role in vision formation. These results were further supported by quantitative PCR analysis, which revealed a significant and continuous (3- to 50-fold) increase in the expression of various voltage-gated channels concomitantly with the increase in the expression of the photoreceptor marker CRX. Conclusions: These results can shed light on the electrophysiologic maturation of neurons in general and PRP in particular and can form the basis for devising and optimizing cell replacement-based vision restoration strategies.

Carbon nanostructures as a scaffold for human embryonic stem cell differentiation toward photoreceptor precursors.

Carbon nanomaterials have been introduced as a scaffold for various biological applications due to their unique physical and electrical properties. Here we studied carbon nanotubes (CNTs) and carbon nanofibers (CNFs) as scaffold materials for the differentiation of human embryonic stem cells (hESCs) towards photoreceptor precursor cells (PRPs). We report on their cytoxicity, their effect on cell morphology, cell-surface interface and the differentiation process. To this end, hESCs were differentiated into PRPs on carbon nanofibers (CNFs), long horizontal CNTs (LHCNTs), vertically aligned CNTs (VACNTs) or glass (control) surfaces. The differentiated cells were investigated by immunohistochemistry, fluorescence imaging and electron microscopy. Our results revealed that the investigated nanomaterials were not cytotoxic to the cells during the differentiation process. The surface interface effect on the cells was apparent, affecting cell directionality, migration and morphology. Interestingly, cell fate was not dependent on the substrate type, as inferred from the similar dynamics of the loss of pluripotency and the comparable expression levels of the photoreceptor marker Crx for all investigated substrates. These results are important for better understanding the effect of nanomaterial surface interaction with differentiating neural cells in general, and for future use of these materials as scaffolds for differentiating photoreceptors for vision restoration in particular.

SEM/FIB Imaging for Studying Neural Interfaces.

Tissue and neural engineering for various regenerative therapies are rapidly growing fields. Of major interest is studying the complex interface between cells and various 3D structures by scanning electron microscopy with focused ion beam. Notwithstanding its unrivaled resolution, the optimal fixation, dehydration, and staining protocols of the samples while preserving the complex cell interface in its natural form, are highly challenging. The aim of this work was to compare and optimize staining and sample drying procedures in order to preserve the cells in their "life-like state" for studying the cell interface with either 3D well-like structures or gold-coated mushroom-shaped electrodes. The process involved chemical fixation using a combination of glutaraldehyde and formaldehyde, followed by gentle drying techniques in which we compared four methods: (critical point drying, hexamethyldisiloxane, repeats of osmium tetroxide-thiocarbohydrazide [OTOTO], and resin) in order to determine the method that best preserves the cell and cell interface morphology. Finally, to visualize the intracellular organelles and membrane, we compared the efficacy of four staining techniques: osmium tetroxide, osmium tetroxide and salts, osmium and uranyl acetate, and OTOTO. Experiments were performed on embryonic stem cell-derived photoreceptor precursors, neural cells, and a human retinal pigment epithelial cell line, which revealed that the optimal processing combination was resin drying and OTOTO staining, as manifested by preservation of cell morphology, the lowest percentage of cellular protrusion breakage as well as a high-quality image. The obtained results pave the way for better understanding the cell interface with various structures for enhancing various biomedical applications.

Cortical Interactions between Prosthetic and Natural Vision.

Outer retinal degenerative diseases, such as retinitis pigmentosa (RP) and age-related macular degeneration (AMD), are among the leading causes of incurable blindness in the Western world [1]. Retinal prostheses have been shown to restore some useful vision by electrically stimulating the remaining retinal neurons [2]. In contrast to inherited retinal degenerative diseases (e.g., RP), typically leading to a complete loss of the visual field, in AMD patients the disease is localized to the macula, leaving the peripheral vision intact. Implanting a retinal prosthesis in the central macula in AMD patients [3, 4] leads to an intriguing situation where the patient's central retina is stimulated electrically, whereas the peripheral healthy retina responds to natural light stimulation. An important question is whether the visual cortex responds to these two concurrent stimuli similarly to the interaction between two adjacent natural light stimuli projected onto healthy retina. Here, we investigated the cortical interactions between prosthetic and natural vision based on visually evoked potentials (VEPs) recorded in rats implanted with photovoltaic subretinal implants. Using this model, where prosthetic and natural vision information are combined in the visual cortex, we observed striking similarities in the interactions of natural and prosthetic vision, including similar effect of background illumination, linear summation of non-patterned stimuli, and lateral inhibition with spatial patterns [5], which increased with target contrast. These results support the idea of combined prosthetic and natural vision in restoration of sight for AMD patients.

High-resolution VSDI retinotopic mapping via a DLP-based projection system.

High-resolution recording of visual cortex activity is an important tool for vision research. Using a customized digital mirror device (DMD) - based system equipped with retinal imaging, we projected visual stimuli directly on the rat retina and recorded cortical responses by voltage-sensitive dye imaging. We obtained robust cortical responses and generated high-resolution retinotopic maps at an unprecedented retinal resolution of 4.6 degrees in the field of view, while further distinguishing between normal and pathological retinal areas. This system is a useful tool for studying the cortical response to localized retinal stimulation and may shed light on various cortical plasticity processes.

Gold nanoparticles for multimodal high-resolution imaging of transplanted cells for retinal replacement therapy.

Aim: Longitudinal tracking of transplanted cells in clinical and experimental setups is crucial for evaluating the efficiency of retinal cell replacement therapies. Materials & methods: Gold nanoparticle-labeled photoreceptor precursors were transplanted in the vitreous and subretinal space of rats and were longitudinally tracked for over a month using optical coherence tomography, computed tomography and fluorescence fundus imaging. Results: This multimodal imaging approach enabled high-resolution long-term tracking and estimation of cell survival in the retina and vitreous, while displaying no toxic effects on the cells or the retina. Conclusion: These observations highlight the applicability of using gold nanoparticle for retinal cell tracking in existing experimental settings and its translational potential for providing more efficient retinal cell therapy in humans.

Active photonic sensing for super-resolved reading performance in simulated prosthetic vision.

In this work, we study the enhancement of simulated prosthetic reading performance through "active photonic sensing" in normally sighted subjects. Three sensing paradigms were implemented: active sensing, in which the subject actively scanned the presented words using the computer mouse, with an option to control text size; passive scanning produced by software-initiated horizontal movements of words; and no scanning. Our findings reveal a 30% increase in word recognition rate with active scanning as compared to no or passive scanning and up to 14-fold increase with zooming. These results highlight the importance of a patient interactive interface and shed light on techniques that can greatly enhance prosthetic vision quality.

An optimized protocol for generating labeled and transplantable photoreceptor precursors from human embryonic stem cells.

Cell replacement therapy is a promising approach for treatment of retinal degenerative diseases. Several protocols for the generation of photoreceptor precursors (PRP) from human embryonic stem cells (hESC) have been reported with variable efficiency. Herein, we show the advantages of use of size-controlled embryoid bodies in the ESC differentiation process using two differentiation protocols. We further explored cell-labeling methods for following the survival of PRP transplanted subretinally in rat eyes. Size-controlled embryoid bodies (EBs) generated using microwell dishes and non-size-controlled EBs generated using V-shaped 96-well plates were differentiated into PRP using two differentiation protocols. The differentiation protocols utilized two different combinations of growth factors. The first, Dkk1, Noggin, and IGF1, and the second protocol used IWR1e, SAG, and CHIR99021. Differentiation efficiency to PRP was analyzed by qPCR, immunocytochemistry, and fluorescence-assisted cell sorting (FACS). Size-controlled IWR1e yielded a significantly higher percent (86.4%) of PRP cells expressing CRX, compared with non-size-controlled IWR1e (51.4%, P = 0.026) or the size-controlled DKK1 protocol (70.5%, p = 0.007). In addition, the IWR1e differentiated cells exhibited a significantly higher fluorescence intensity of CRX immunostaining, compared with the DKK1 protocol, consistent with higher protein expression levels. The IWR1e cells exhibited higher maturation levels, as manifested by lower early neuronal marker PAX6 and pluripotency marker OCT4 levels compared with the DKK1 protocol. The expression of other late photoreceptor markers (NRL, recoverin) were similar among the differentiation groups. PRP cells were labeled by using hESC constitutively expressing EGFP or by AAV-GFP transduction. Finally, we transplanted the cells in the subretinal space of wild-type rats and monitored their survival over several weeks. The AAV2 serotype efficiently transduced the PRP cells, whereas other serotypes yielded low or no transduction. Following subretinal transplantation of GFP-labeled PRP, 63% of the cells were detected at 4 weeks post-transplantation. In conclusion, we show here that the IWR1e protocol using size-controlled EBs efficiently generated of PRP that could be labeled and followed in-vivo for weeks. The data from this study is an advance toward the goal of PRP transplantation therapy for retinal degenerative diseases.

Endovascular Electrical Stimulation - A Novel Hemorrhage Control Technique.

OBJECTIVE: In this study we present a novel approach for inducing vasoconstriction by pulsed electrical treatment delivered via endovascular electrodes, which can be used in cases where external access to the vessel is limited. METHODS: Using computer simulations, we optimized various geometries of endovascular electrodes to maximize the induced electric field on the arterial wall. Using the optimal configuration parameters, we investigated endovascular induced vasoconstriction in both the carotid and femoral sheep arteries. RESULTS: Endovascular electrodes induced robust vasoconstriction in the carotid artery of sheep, showing gradual recovery following treatment. Moreover, the obtained vasoconstriction was accompanied by a sevenfold decrease in blood loss for 100% constriction, compared with no treatment (6ml vs 42ml, p<0.001). The femoral artery was less amenable to the electrical treatment, which we hypothesize results from the reduced density of the sympathetic system's innervation of the adventitia of the sheep femoral artery, as was validated by immunohistochemical analysis. Finally, treatment safety was validated through arterial histological studies, in which no adverse effect was observed, and through computer modeling, which depicted a negligible temperature increase. SIGNIFICANCE: These results are an important step toward developing a novel approach for inducing reversible and controlled vasoconstriction in arteries that are remote from access.

Evaluation of Critical Flicker-Fusion Frequency Measurement Methods for the Investigation of Visual Temporal Resolution.

Recent studies highlight the importance of the temporal domain in visual processing. Critical Flicker-Fusion Frequency (CFF), the frequency at which a flickering light is perceived as continuous, is widely used for evaluating visual temporal processing. However, substantial variability in the psychophysical paradigms, used for measuring CFF, leads to substantial variability in the reported results. Here, we report on a comprehensive comparison of CFF measurements through three different psychophysical paradigms: methods of limits; method of constant stimuli, and staircase method. Our results demonstrate that the CFF can be reliably measured with high repeatability by all three psychophysics methods. However, correlations (r = 0.92, p≪0.001) and agreement (Bland Altman test indicated 95% confidence limit variation of ±3.6 Hz), were highest between the staircase and the constant stimuli methods. The time required to complete the test was significantly longer for the constant stimuli method as compared to other methods (p < 0.001). Our results highlight the suitability of the adaptive paradigm for efficiently measuring temporal resolution in the visual system.

Head mounted DMD based projection system for natural and prosthetic visual stimulation in freely moving rats.

Novel technologies are constantly under development for vision restoration in blind patients. Many of these emerging technologies are based on the projection of high intensity light patterns at specific wavelengths, raising the need for the development of specialized projection systems. Here we present and characterize a novel projection system that meets the requirements for artificial retinal stimulation in rats and enables the recording of cortical responses. The system is based on a customized miniature Digital Mirror Device (DMD) for pattern projection, in both visible (525 nm) and NIR (915 nm) wavelengths, and a lens periscope for relaying the pattern directly onto the animal's retina. Thorough system characterization and the investigation of the effect of various parameters on obtained image quality were performed using ZEMAX. Simulation results revealed that images with an MTF higher than 0.8 were obtained with little effect of the vertex distance. Increased image quality was obtained at an optimal pupil diameter and smaller field of view. Visual cortex activity data was recorded simultaneously with pattern projection, further highlighting the importance of the system for prosthetic vision studies. This novel head mounted projection system may prove to be a vital tool in studying natural and artificial vision in behaving animals.

Endovascular Electrodes for Electrical Stimulation of Blood Vessels for Vasoconstriction - a Finite Element Simulation Study.

Hemorrhagic shock accounts for 30-40 percent of trauma mortality, as bleeding may sometimes be hard to control. Application of short electrical pulses on blood vessels was recently shown to elicit robust vasoconstriction and reduction of blood loss following vascular injury. In this study we present a novel approach for vasoconstriction based on endovascular application of electrical pulses for situations where access to the vessel is limited. In addition to ease of access, we hypothesize that this novel approach will result in a localized and efficient vasoconstriction. Using computer modeling (COMSOL Multiphysics, Electric Currents Module), we studied the effect of endovascular pulsed electrical treatment on abdominal aorta of pigs, and compared the efficiency of different electrodes configurations on the electric field amplitude, homogeneity and locality when applied on a blood vessel wall. Results reveal that the optimal configuration is the endovascular approach where four electrodes are used, spaced 13 mm apart. Furthermore, computer based temperature investigations (bio-heat model, COMSOL Multiphysics) show that the maximum expected temperature rise is of 1.2 degrees; highlighting the safety of the four endovascular electrodes configuration. These results can aid in planning the application of endovascular pulsed electrical treatment as an efficient and safe vasoconstriction approach.