Suppression of natural lens fluorescence in fundus autofluorescence measurements: review of hardware solutions.

Fluorescence lifetime imaging ophthalmoscopy (FLIO), a technique for investigating metabolic changes in the eye ground, can reveal the first signs of diseases related to metabolism. The fluorescence of the natural lens overlies the fundus fluorescence. Although the influence of natural lens fluorescence can be somewhat decreased with mathematical models, excluding this influence during the measurement by using hardware enables more exact estimation of the fundus fluorescence. Here, we analyze four 1-photon excitation hardware solutions to suppress the influence of natural lens fluorescence: aperture stop separation, confocal scanning laser ophthalmoscopy, combined confocal scanning laser ophthalmoscopy and aperture stop separation, and dual point confocal scanning laser ophthalmoscopy. The effect of each principle is demonstrated in examples. The best suppression is provided by the dual point principle, realized with a confocal scanning laser ophthalmoscope. In this case, in addition to the fluorescence of the whole eye, the fluorescence of the anterior part of the eye is detected from a non-excited spot of the fundus. The intensity and time-resolved fluorescence spectral data of the fundus are derived through the subtraction of the simultaneously measured fluorescence of the excited and non-excited spots. Advantages of future 2-photon fluorescence excitation are also discussed. This study provides the first quantitative evaluation of hardware principles to suppress the fluorescence of the natural lens during measurements of fundus autofluorescence.

Evaluating Current Density Modeling of Non-Invasive Eye and Brain Electrical Stimulation Using Phosphene Thresholds.

Because current flow cannot be measured directly in the intact retina or brain, current density distribution models were developed to estimate it during magnetic or electrical stimulation. A paradigm is now needed to evaluate if current flow modeling can be related to physiologically meaningful signs of true current distribution in the human brain. We used phosphene threshold measurements (PTs) as surrogate markers of current-flow to determine if PTs, evoked by transcranial alternating current stimulation (tACS), can be matched with current density estimates generated by head model-based computer simulations. Healthy, male subjects (n=15) were subjected to three-staged PT measurements comparing six unilateral and one bilateral stimulation electrode montages according to the 10/20 system: Fp2-Suborbital right (So), Fp2-right shoulder (rS), Fp2-Cz, Fp2- O2, So-rS, Cz-F8 and F7-F8. The stimulation frequency was set at 16 Hz. Subjects were asked to report the appearance and localization of phosphenes in their visual field for every montage. Current density models were built using multi-modal imaging data of a standard brain, meshed with isotropic conductivities of different tissues of the head using the SimBio and SCIRun software packages. We observed that lower PTs were associated with higher simulated current levels in the unilateral montages of the model head, and shorter electrode distances to the eye had lower PTs. The lowest mean PT and the lowest variability were found in the F7-F8 montage ( [Formula: see text]). Our results confirm the hypothesis that phosphenes are primarily of retinal origin, and they provide the first in vivo evidence that computer models of current flow using head models are a valid tool to estimate real current flow in the human eye and brain.

Comparison of algorithms to suppress artifacts from the natural lens in fluorescence lifetime imaging ophthalmoscopy (FLIO).

Fluorescence lifetime imaging ophthalmoscopy (FLIO) has developed as a new diagnostic tool in ophthalmology. FLIO measurements are taken from 30° retinal fields in two spectral channels (short spectral channel (SSC): 498-560 nm, long spectral channel (LSC): 560-720 nm). Because of the layered structure of the eye, the detected signal is an interaction of the fluorescence decay of the anterior part and of the fundus. By comparing FLIO measurements before and after cataract surgery, the impact of the natural lens was proven, despite the application of a confocal laser scanning (cSLO) technique. The goal of this work was to determine the best algorithmic solution to isolate the sole fundus fluorescence lifetime from the measured signal, suppressing artifacts from the natural lens. Three principles based on a tri-exponential model were investigated: a tailfit, a layer-based approach with a temporally shifted component, and the inclusion of a separately measured fluorescence decay of the natural lens. The mean fluorescence lifetime τm,12 is calculated using only the shortest and the intermediate exponential component. τm,all is calculated using all three exponential components. The results of tri-exponential tailfit after cataract surgery were considered as a reference, because the implanted artificial lens can be assumed as non-fluorescent. In SSC, the best accordance of τm,all of the reference was determined with τm,12 of the tailfit before surgery. If high-quality natural lens measurements are available, the correspondence of τm,12 is best with τm,all of the reference. In LSC, there is a good accordance for all models between τm,12 before and after surgery. To study the pure fundus fluorescence decay in eyes with natural lenses, we advise to utilize fluorescence lifetime τm,12 of a triple-exponential tailfit, as it corresponds well with the mean fluorescence lifetime τm,all of eyes with fluorescence-less artificial intraocular lenses.

Low intensity transcranial electric stimulation: Safety, ethical, legal regulatory and application guidelines.

Low intensity transcranial electrical stimulation (TES) in humans, encompassing transcranial direct current (tDCS), transcutaneous spinal Direct Current Stimulation (tsDCS), transcranial alternating current (tACS), and transcranial random noise (tRNS) stimulation or their combinations, appears to be safe. No serious adverse events (SAEs) have been reported so far in over 18,000 sessions administered to healthy subjects, neurological and psychiatric patients, as summarized here. Moderate adverse events (AEs), as defined by the necessity to intervene, are rare, and include skin burns with tDCS due to suboptimal electrode-skin contact. Very rarely mania or hypomania was induced in patients with depression (11 documented cases), yet a causal relationship is difficult to prove because of the low incidence rate and limited numbers of subjects in controlled trials. Mild AEs (MAEs) include headache and fatigue following stimulation as well as prickling and burning sensations occurring during tDCS at peak-to-baseline intensities of 1-2mA and during tACS at higher peak-to-peak intensities above 2mA. The prevalence of published AEs is different in studies specifically assessing AEs vs. those not assessing them, being higher in the former. AEs are frequently reported by individuals receiving placebo stimulation. The profile of AEs in terms of frequency, magnitude and type is comparable in healthy and clinical populations, and this is also the case for more vulnerable populations, such as children, elderly persons, or pregnant women. Combined interventions (e.g., co-application of drugs, electrophysiological measurements, neuroimaging) were not associated with further safety issues. Safety is established for low-intensity 'conventional' TES defined as <4mA, up to 60min duration per day. Animal studies and modeling evidence indicate that brain injury could occur at predicted current densities in the brain of 6.3-13A/m2 that are over an order of magnitude above those produced by tDCS in humans. Using AC stimulation fewer AEs were reported compared to DC. In specific paradigms with amplitudes of up to 10mA, frequencies in the kHz range appear to be safe. In this paper we provide structured interviews and recommend their use in future controlled studies, in particular when trying to extend the parameters applied. We also discuss recent regulatory issues, reporting practices and ethical issues. These recommendations achieved consensus in a meeting, which took place in Göttingen, Germany, on September 6-7, 2016 and were refined thereafter by email correspondence.

[Flexible Guidance of Ultra-Short Laser Pulses in Ophthalmic Therapy Systems]. / Flexible Führung ultrakurzer Laserpulse in ophthalmologischen Therapiesystemen.

In the last 20 years, the role of ultrashort pulsed lasers in ophthalmology has become increasingly important. However, it is still impossible to guide ultra-short laser pulses with standard glass fibres. The highly energetic femtosecond pulses would destroy the fibre material, and non-linear dispersion effects would significantly change beam parameters. In contrast, photonic crystal fibres mainly guide the laser pulses in air, so that absorption and dispersive pulse broadening have essentially no effect. This article compares classical beam guidance with mirrors, lenses and prisms with photonic crystal fibres and describes the underlying concepts and the current state of technology. A classical mirror arm possesses more variable optical properties, while the HCF (Hollow-Core Photonic Crystal Fibre) must be matched in terms of the laser energy and the laser spectrum. In contrast, the HCF has more advantages in respect of handling, system integration and costs. For applications based on photodisruptive laser-tissue interaction, the relatively low damage threshold of photonic crystal fibres compared to classic beam guiding systems is unacceptable. If, however, pulsed laser radiation has a sufficiently low peak intensity, e.g. as used for plasma-induced ablation, photonic crystal fibres can definitely be considered as an alternative solution to classic beam guidance.

EEG and MEG: sensitivity to epileptic spike activity as function of source orientation and depth.

Simultaneous electroencephalography (EEG) and magnetoencephalography (MEG) recordings of neuronal activity from epileptic patients reveal situations in which either EEG or MEG or both modalities show visible interictal spikes. While different signal-to-noise ratios (SNRs) of the spikes in EEG and MEG have been reported, a quantitative relation of spike source orientation and depth as well as the background brain activity to the SNR has not been established. We investigated this quantitative relationship for both dipole and patch sources in an anatomically realistic cortex model. Altogether, 5600 dipole and 3300 patch sources were distributed on the segmented cortical surfaces of two volunteers. The sources were classified according to their quantified depths and orientations, ranging from 20 mm to 60 mm below the skin surface and radial and tangential, respectively. The source time-courses mimicked an interictal spike, and the simulated background activity emulated resting activity. Simulations were conducted with individual three-compartment boundary element models. The SNR was evaluated for 128 EEG, 102 MEG magnetometer, and 204 MEG gradiometer channels. For superficial dipole and superficial patch sources, EEG showed higher SNRs for dominantly radial orientations, and MEG showed higher values for dominantly tangential orientations. Gradiometers provided higher SNR than magnetometers for superficial sources, particularly for those with dominantly tangential orientations. The orientation dependent difference in SNR in EEG and MEG gradually changed as the sources were located deeper, where the interictal spikes generated higher SNRs in EEG compared to those in MEG for all source orientations. With deep sources, the SNRs in gradiometers and magnetometers were of the same order. To better detect spikes, both EEG and MEG should be used.

Modular multipin electrodes for comfortable dry EEG.

Electrode and cap concepts for continuous and ubiquitous monitoring of brain activity will open up new fields of application and contribute to increased use of electroencephalography (EEG) in clinical routine, neurosciences, brain-computer-interfacing and out-of-the-lab monitoring. However, mobile and unobtrusive applications are currently hindered by the lack of applicable convenient and reliable electrode and cap systems. We propose a novel modular electrode concept based on a flexible polymer substrate, coated with electrically conductive metallic films. The overall concept enables design adaptation to different head regions and cap designs. We describe the single modules of the system and investigate the influence of electrode pin number, coating material and adduction force on electrode-skin impedance and perceived wearing comfort. Our results contribute to rapid and comfortable multichannel dry EEG.

Development of a vector-tensor system to measure the absolute magnetic flux density and its gradient in magnetically shielded rooms.

Several experiments in fundamental physics demand an environment of very low, homogeneous, and stable magnetic fields. For the magnetic characterization of such environments, we present a portable SQUID system that measures the absolute magnetic flux density vector and the gradient tensor. This vector-tensor system contains 13 integrated low-critical temperature (LTc) superconducting quantum interference devices (SQUIDs) inside a small cylindrical liquid helium Dewar with a height of 31 cm and 37 cm in diameter. The achievable resolution depends on the flux density of the field under investigation and its temporal drift. Inside a seven-layer mu-metal shield, an accuracy better than ±23 pT for the components of the static magnetic field vector and ±2 pT/cm for each of the nine components of the gradient tensor is reached by using the shifting method.

Novel Multipin Electrode Cap System for Dry Electroencephalography.

Current usage of electroencephalography (EEG) is limited to laboratory environments. Self-application of a multichannel wet EEG caps is practically impossible, since the application of state-of-the-art wet EEG sensors requires trained laboratory staff. We propose a novel EEG cap system with multipin dry electrodes overcoming this problem. We describe the design of a novel 24-pin dry electrode made from polyurethane and coated with Ag/AgCl. A textile cap system holds 97 of these dry electrodes. An EEG study with 20 volunteers compares the 97-channel dry EEG cap with a conventional 128-channel wet EEG cap for resting state EEG, alpha activity, eye blink artifacts and checkerboard pattern reversal visual evoked potentials. All volunteers report a good cap fit and good wearing comfort. Average impedances are below 150 kΩ for 92 out of 97 dry electrodes, enabling recording with standard EEG amplifiers. No significant differences are observed between wet and dry power spectral densities for all EEG bands. No significant differences are observed between the wet and dry global field power time courses of visual evoked potentials. The 2D interpolated topographic maps show significant differences of 3.52 and 0.44% of the map areas for the N75 and N145 VEP components, respectively. For the P100 component, no significant differences are observed. Dry multipin electrodes integrated in a textile EEG cap overcome the principle limitations of wet electrodes, allow rapid application of EEG multichannel caps by non-trained persons, and thus enable new fields of application for multichannel EEG acquisition.

Primary somatosensory contextual modulation is encoded by oscillation frequency change.

OBJECTIVE: This study characterized thalamo-cortical communication by assessing the effect of context-dependent modulation on the very early somatosensory evoked high-frequency oscillations (HF oscillations). METHODS: We applied electrical stimuli to the median nerve together with an auditory oddball paradigm, presenting standard and deviant target tones representing differential cognitive contexts to the constantly repeated electrical stimulation. Median nerve stimulation without auditory stimulation served as unimodal control. RESULTS: A model consisting of one subcortical (near thalamus) and two cortical (Brodmann areas 1 and 3b) dipolar sources explained the measured HF oscillations. Both at subcortical and the cortical levels HF oscillations were significantly smaller during bimodal (somatosensory plus auditory) than unimodal (somatosensory only) stimulation. A delay differential equation model was developed to investigate interactions within the 3-node thalamo-cortical network. Importantly, a significant change in the eigenfrequency of Brodmann area 3b was related to the context-dependent modulation, while there was no change in the network coupling. CONCLUSION: This model strongly suggests cortico-thalamic feedback from both cortical Brodmann areas 1 and 3b to the thalamus. With the 3-node network model, thalamo-cortical feedback could be described. SIGNIFICANCE: Frequency encoding plays an important role in contextual modulation in the somatosensory thalamo-cortical network.

Induced schizophrenic like breathing pattern leads to impaired cardiorespiratory coupling in healthy subjects.

Schizophrenia is referred to as one of the most severe mental disorders in the world, and patients with this condition are associated with high cardiac mortality rates. However, the reasons for this high mortality rates are still under debate. One major contributing factor seems to be that a dysfunction of the autonomic nervous system (ANS) is evident in schizophrenia. Thereby, recent investigations focused on the analyses of respiration and cardiorespiratory coupling (CRC) in these patients.The objective of this study was to characterize the causal strength and direction of CRC applying the normalized short time partial directed coherence (NSTPDC) approach in healthy subjects (CON). In this study 25 healthy control subjects were enrolled matching in terms of age and gender to schizophrenic patients. CON were measured in resting condition (pre), breathing with schizophrenic like breathing pattern (stress) and resting condition for recovery (post). During stress CON were asked to breathe with a controlled fixed respiratory frequency, inspirationand expiration time. We found during induced stress significantly increased heart rate and reduced heart rate variability, increased breathing rate and reduced respiratory variability as well as impaired CRC for CON. In conclusion, during induced pathological breathing pattern CON exhibit an impaired and altered heart rate and respiratory regulation and CRC as indicators of a vagal withdrawal and sympathetic overdrive by the ANS. CRC analyses revealed impaired coupling strength and direction, with a driver-responder relationship from respiration to heart rate during stress. These findings support the central role of the respiratory dysregulation in schizophrenia and the assumed suppression of higher regulatory centers of the brain stem due to arousals and permanent stress situations in acute schizophrenia.

Quantitative imaging of magnetic nanoparticles by magnetorelaxometry with multiple excitation coils.

New therapies against cancer based on magnetic nanoparticles (MNPs) require a quantitative spatially resolved imaging of MNPs inside a body. In magnetorelaxometry (MRX), a distribution of nanoparticles can be quantified non-invasively by measuring its relaxation after removal of an external magnetizing field. Conventionally, in MRX the sample is exposed to a homogeneous magnetizing field resulting in a quantitative reconstruction with rather poor spatial resolution. Theoretical work suggests an improvement of spatial resolution may be achieved by a sequential application of inhomogeneous fields magnetizing only parts of a sample. Here, we experimentally demonstrate the feasibility of this approach by reconstructing a nanoparticle distribution inside a compact three-dimensional volume phantom made of 54 gypsum cubes (1 cm(3) cube(-1)), of which 12 gypsum cubes were filled with MNPs. Using 48 small excitation coils surrounding the phantom, a sequence of MRX signals was obtained where only those MNPs near an individual coil contribute. By combined evaluation of these 48 MRX measurements, the positions and content of the 12 MNP-filled cubes could be determined accurately with a deviation below 4%, while by conventional homogeneous MRX only the MNP content was reconstructable with a deviation of about 9%. The results demonstrate the improvement of quantitative MRX imaging by using sequential activation of multiple magnetizing fields.

Magnetoencephalography signals are influenced by skull defects.

OBJECTIVE: Magnetoencephalography (MEG) signals had previously been hypothesized to have negligible sensitivity to skull defects. The objective is to experimentally investigate the influence of conducting skull defects on MEG and EEG signals. METHODS: A miniaturized electric dipole was implanted in vivo into rabbit brains. Simultaneous recording using 64-channel EEG and 16-channel MEG was conducted, first above the intact skull and then above a skull defect. Skull defects were filled with agar gels, which had been formulated to have tissue-like homogeneous conductivities. The dipole was moved beneath the skull defects, and measurements were taken at regularly spaced points. RESULTS: The EEG signal amplitude increased 2-10 times, whereas the MEG signal amplitude reduced by as much as 20%. The EEG signal amplitude deviated more when the source was under the edge of the defect, whereas the MEG signal amplitude deviated more when the source was central under the defect. The change in MEG field-map topography (relative difference measure, RDM(∗)=0.15) was geometrically related to the skull defect edge. CONCLUSIONS: MEG and EEG signals can be substantially affected by skull defects. SIGNIFICANCE: MEG source modeling requires realistic volume conductor head models that incorporate skull defects.

Novel flexible Dry multipin electrodes for EEG: Signal quality and interfacial impedance of Ti and TiN coatings.

Conventional Silver/Silver-Chloride electrodes are inappropriate for routine high-density EEG and emerging new fields of application like brain computer interfaces. A novel multipin electrode design is proposed. It enables rapid and easy application while maintaining signal quality and patient comfort. The electrode design is described and impedance and EEG tests are performed with Titanium and Titanium Nitride coated electrodes. The results are compared to conventional reference electrodes in a multi-volunteer study. The calculated signal parameters prove the multipin electrode concept to reproducibly acquire EEG signal quality comparable to Ag/AgCl electrodes. The promising results encourage further investigation and can provide a technological base for future preparation-free multichannel EEG systems.

Time-frequency mixed-norm estimates: sparse M/EEG imaging with non-stationary source activations.

Magnetoencephalography (MEG) and electroencephalography (EEG) allow functional brain imaging with high temporal resolution. While solving the inverse problem independently at every time point can give an image of the active brain at every millisecond, such a procedure does not capitalize on the temporal dynamics of the signal. Linear inverse methods (minimum-norm, dSPM, sLORETA, beamformers) typically assume that the signal is stationary: regularization parameter and data covariance are independent of time and the time varying signal-to-noise ratio (SNR). Other recently proposed non-linear inverse solvers promoting focal activations estimate the sources in both space and time while also assuming stationary sources during a time interval. However such a hypothesis holds only for short time intervals. To overcome this limitation, we propose time-frequency mixed-norm estimates (TF-MxNE), which use time-frequency analysis to regularize the ill-posed inverse problem. This method makes use of structured sparse priors defined in the time-frequency domain, offering more accurate estimates by capturing the non-stationary and transient nature of brain signals. State-of-the-art convex optimization procedures based on proximal operators are employed, allowing the derivation of a fast estimation algorithm. The accuracy of the TF-MxNE is compared with recently proposed inverse solvers with help of simulations and by analyzing publicly available MEG datasets.