Three-micrometer-diameter needle electrode with an amplifier for extracellular in vivo recordings.

Microscale needle-electrode devices offer neuronal signal recording capability in brain tissue; however, using needles of smaller geometry to minimize tissue damage causes degradation of electrical properties, including high electrical impedance and low signal-to-noise ratio (SNR) recording. We overcome these limitations using a device assembly technique that uses a single needle-topped amplifier package, called STACK, within a device of ∼1 × 1 mm2 Based on silicon (Si) growth technology, a <3-µm-tip-diameter, 400-µm-length needle electrode was fabricated on a Si block as the module. The high electrical impedance characteristics of the needle electrode were improved by stacking it on the other module of the amplifier. The STACK device exhibited a voltage gain of >0.98 (-0.175 dB), enabling recording of the local field potential and action potentials from the mouse brain in vivo with an improved SNR of 6.2. Additionally, the device allowed us to use a Bluetooth module to demonstrate wireless recording of these neuronal signals; the chronic experiment was also conducted using STACK-implanted mice.

Robust brightness enhancement across a luminance range of the glare illusion.

The glare illusion refers to brightness enhancement and the perception of a self-luminous appearance that occurs when a central region is surrounded by a luminance gradient. The center region appears to be a light source, with its light dispersing into the surrounding region. If the luminous edge is critical for generating the illusion, modulating the perceived luminance of the image, and switching its appearance from luminous to nonluminous, would have a strong impact on lightness and brightness estimation. Here, we quantified the illusion in two ways, by assessing brightness enhancement and examining whether the center region appeared luminous. Thus, we could determine whether the two effects occurred jointly or independently. We examined a wide luminance range of center regions, from 0 to 200% relative to background. Brightness enhancement in the illusion was observed for a wide range of luminances (20% to 200% relative to background), while a luminous-white appearance was observed when the center region luminance was 145% of the background. These results exclude the possibility that brightness enhancement occurs because the stimuli appear self-luminous. We suggest that restoring the original image intensity precedes the perceptual process of lightness estimation.

Color vision test for dichromatic and trichromatic macaque monkeys.

Dichromacy is a color vision defect in which one of the three cone photoreceptors is absent. Individuals with dichromacy are called dichromats (or sometimes "color-blind"), and their color discrimination performance has contributed significantly to our understanding of color vision. Macaque monkeys, which normally have trichromatic color vision that is nearly identical to humans, have been used extensively in neurophysiological studies of color vision. In the present study we employed two tests, a pseudoisochromatic color discrimination test and a monochromatic light detection test, to compare the color vision of genetically identified dichromatic macaques (Macaca fascicularis) with that of normal trichromatic macaques. In the color discrimination test, dichromats could not discriminate colors along the protanopic confusion line, though trichromats could. In the light detection test, the relative thresholds for longer wavelength light were higher in the dichromats than the trichromats, indicating dichromats to be less sensitive to longer wavelength light. Because the dichromatic macaque is very rare, the present study provides valuable new information on the color vision behavior of dichromatic macaques, which may be a useful animal model of human dichromacy. The behavioral tests used in the present study have been previously used to characterize the color behaviors of trichromatic as well as dichromatic new world monkeys. The present results show that comparative studies of color vision employing similar tests may be feasible to examine the difference in color behaviors between trichromatic and dichromatic individuals, although the genetic mechanisms of trichromacy/dichromacy is quite different between new world monkeys and macaques.

A Fine-Scale and Minimally Invasive Marking Method for Use with Conventional Tungsten Microelectrodes.

In neurophysiology, achieving precise correlation between physiological responses and anatomic structures is a significant challenge. Therefore, the accuracy of the electrode marking method is crucial. In this study, we describe a tungsten-deposition method, in which tungsten oxide is generated by applying biphasic current pulses to conventional tungsten electrodes. The electrical current used was 40-50 µA, which is similar to that used in electrical microstimulation experiments. The size of the markings ranged from 10 to 100 µm, corresponding to the size of the electrode tip, which is smaller than that of existing marking methods. Despite the small size of the markings, detection is easy as the marking appears in bright red under dark-field observation after Nissl staining. This marking technique resulted in low tissue damage and was maintained in vivo for at least two years. The feasibility of this method was tested in mouse and macaque brains.

Low-invasive neural recording in mouse models with diabetes via an ultrasmall needle-electrode.

Diabetes is known to cause a variety of complications, having a high correlation with Alzheimer's disease. Electrophysiological recording using a microscale needle electrode is a promising technology for the study, however, diabetic brain tissue is more difficult to record neuronal activities than normal tissue due to these complications including the development of cerebrovascular disease. Here we show an electrophysiological methodology for diabetic db/db mice (+Leprdb/+Leprdb) using a 4-µm-tip diameter needle-electrode device. The needle electrode minimized the tissue injury when compared to a typical larger metal electrode, as confirmed by bleeding during penetration. The proposed electrode device showed both acute and chronic in vivo recording capabilities for diabetic mice while reducing the glial cells' responses. Because of these device characteristics, the 4-µm-tip diameter needle-electrode will allow electrophysiological studies on diabetes models of not only mice, as proven in this study, but also other animals.

Influence of Stimulus Size on Simultaneous Chromatic Induction.

Chromatic induction is a major contextual effect of color appearance. Patterned backgrounds are known to induce strong chromatic induction effects. However, it has not been clarified whether the spatial extent of the chromatic surrounding induces a chromatic contrast or assimilation effects. In this study, we examined the influence of the width of a center line and its flanking white contour on the color appearance when the line was surrounded by chromatic backgrounds. A strong color shift was observed when the center line was flanked by white contours with the L/M- and S-cone chromatic backgrounds. There was a difference between the optimal widths of the center line and the contour for the shift in color appearance for the L/M-cone chromaticity (0.9 and 1.1-1.7 min, respectively) and the S-cone chromaticity (8.2-17.5 and 0.9-2.5 min, respectively). The optimal width of the center line for the L/M-cone was finer than the resolution-limit width of the chromatic contrast sensitivity and coarser than that of the luminance contrast sensitivity. Thus, the color appearance of the center line could be obtained by integrating broad chromatic information and fine luminance details. Due to blurring and chromatic aberrations, the simulated artifact was large for the darker center line and S-cone background, thus suggesting that the artifact could explain the luminance dependency of the induction along the S-cone chromaticity. Moreover, the findings of this study reveal that the dominant factor of the color shift is neural instead of optical.

The Role of Specular Reflections and Illumination in the Perception of Thickness in Solid Transparent Objects.

Specular reflections and refractive distortions are complex image properties of solid transparent objects, but despite this complexity, we readily perceive the 3D shapes of these objects (e.g., glass and clear plastic). We have found in past work that relevant sources of scene complexity have differential effects on 3D shape perception, with specular reflections increasing perceived thickness, and refractive distortions decreasing perceived thickness. In an object with both elements, such as glass, the two optical properties may complement each other to support reliable perception of 3D shape. We investigated the relative dominance of specular reflection and refractive distortions in the perception of shape. Surprisingly, the ratio of specular reflection to refractive component was almost equal to that of ordinary glass and ice, which promote correct percepts of 3D shape. The results were also explained by the variance in local RMS contrast in stimulus images but may depend on overall luminance and contrast of the surrounding light field.

A floating 5 µm-diameter needle electrode on the tissue for damage-reduced chronic neuronal recording in mice.

Microelectrode technology is essential in electrophysiology and has made contributions to neuroscience as well as to medical applications. However, it is necessary to minimize tissue damage associated with needle-like electrode on the brain tissue and the implantation surgery, which makes stable chronic recording impossible. Here, we report on an approach for using a 5 µm-diameter needle electrode, which enables the following of tissue motions, via a surgical method. The electrode is placed on the brain tissue of a mouse with a dissolvable material, reducing the physical stress to the tissue; this is followed by the implantation of the electrode device in the brain without fixing it to the cranium, achieving a floating electrode architecture on the tissue. The electrode shows stable recording with no significant degradation of the signal-to-noise ratios for 6 months, and minimized tissue damage is confirmed compared to that when using a cranium-fixed electrode with the same needle geometry.

Nanoneedle-Electrode Devices for In Vivo Recording of Extracellular Action Potentials.

Microscale needle-like electrode technologies offer in vivo extracellular recording with a high spatiotemporal resolution. Further miniaturization of needles to nanoscale minimizes tissue injuries; however, a reduced electrode area increases electrical impedance that degrades the quality of neuronal signal recording. We overcome this limitation by fabricating a 300 nm tip diameter and 200 µm long needle electrode where the amplitude gain with a high-impedance electrode (>15 MΩ, 1 kHz) was improved from 0.54 (-5.4 dB) to 0.89 (-1.0 dB) by stacking it on an amplifier module of source follower. The nanoelectrode provided the recording of both local field potential (<300 Hz) and action potential (>500 Hz) in the mouse cortex, in contrast to the electrode without the amplifier. These results suggest that microelectrodes can be further minimized by the proposed amplifier configuration for low-invasive recording and electrophysiological studies in submicron areas in tissues, such as dendrites and axons.

Advances in color science: from retina to behavior.

Color has become a premier model system for understanding how information is processed by neural circuits, and for investigating the relationships among genes, neural circuits, and perception. Both the physical stimulus for color and the perceptual output experienced as color are quite well characterized, but the neural mechanisms that underlie the transformation from stimulus to perception are incompletely understood. The past several years have seen important scientific and technical advances that are changing our understanding of these mechanisms. Here, and in the accompanying minisymposium, we review the latest findings and hypotheses regarding color computations in the retina, primary visual cortex, and higher-order visual areas, focusing on non-human primates, a model of human color vision.

Categorical properties of the color term "GOLD".

Humans are able to categorize an infinite variety of surface colors into a small number of color terms. Previous studies have shown that 11 basic color terms are commonly used in fully developed languages. These studies usually used flat matte color plates as stimuli, but we can also perceive the colors of glossy surfaces by discounting the effect of the gloss. However, color terms such as GOLD and SILVER are specifically associated with glossy surfaces. In this study, we conducted a categorical color-naming task to examine whether the color terms GOLD and SILVER could be located in a stimulus space defined by combining CIE xy chromaticity coordinates and surface reflectance and whether they had categorical properties like ordinary basic color terms. We found that GOLD and SILVER were used for specific ranges of chromaticities with stimuli having large specular reflectances. Moreover, the strengths of the categorical properties, as assessed using measures of consistency, consensus, and reaction time, were comparable to those of the basic color terms, indicating that GOLD and SILVER are categorical color terms specifically associated with glossy surfaces. This also indicates that humans do not always discount surface gloss to identify colors but can utilize this information to categorize colors.

Effects of task demands on the responses of color-selective neurons in the inferior temporal cortex.

Categorization and fine discrimination are two different functions in visual perception, and we can switch between these two functions depending on the situation or task demands. To explore how visual cortical neurons behave in such situations, we recorded the activities of color-selective neurons in the inferior temporal (IT) cortex of two monkeys trained to perform a color categorization task, a color discrimination task and a simple fixation task. Many IT neurons changed their activity depending upon the task, although color selectivity was well conserved. A majority of neurons showed stronger responses during the categorization task. Moreover, for the population of IT neurons as a whole, signals contributing to performing the categorization task were enhanced. These results imply that judgment of color category by color-selective IT neurons is facilitated during the categorization task and suppressed during the discrimination task as a consequence of task-dependent modulation of their activities.

Relating neuronal firing patterns to functional differentiation of cerebral cortex.

It has been empirically established that the cerebral cortical areas defined by Brodmann one hundred years ago solely on the basis of cellular organization are closely correlated to their function, such as sensation, association, and motion. Cytoarchitectonically distinct cortical areas have different densities and types of neurons. Thus, signaling patterns may also vary among cytoarchitectonically unique cortical areas. To examine how neuronal signaling patterns are related to innate cortical functions, we detected intrinsic features of cortical firing by devising a metric that efficiently isolates non-Poisson irregular characteristics, independent of spike rate fluctuations that are caused extrinsically by ever-changing behavioral conditions. Using the new metric, we analyzed spike trains from over 1,000 neurons in 15 cortical areas sampled by eight independent neurophysiological laboratories. Analysis of firing-pattern dissimilarities across cortical areas revealed a gradient of firing regularity that corresponded closely to the functional category of the cortical area; neuronal spiking patterns are regular in motor areas, random in the visual areas, and bursty in the prefrontal area. Thus, signaling patterns may play an important role in function-specific cerebral cortical computation.

Large enhancement of simultaneous color contrast by white flanking contours.

Simultaneous color contrast and assimilation are mutually opposing effects on color appearance, and their magnitude depends on spatial context. The Monnier-Shevell illusion induces a large color shift by a synergy of simultaneous assimilation and contrast using the alternating color of proximal and distant surrounds. The illusion induces a prominent effect along the blue-yellow color axis, but a subtle effect along the orthogonal color axis. In this study, we report an illusion generated by an extremely thin gray line on a cyan background that appears reddish when the line is flanked by thin white contours. We quantified the color appearance of the gray line in a color matching experiment and found that the color shift of the gray line with white contours induced large color shifts. It is also known that luminance contrast between a center and its surrounds affects the magnitude of simultaneous color contrast. However, our color contrast effects were larger for a dark line rather than for a pale line. In contrast, the perceived color shift of the line without the contours increased as the luminance of the gray line increased, supporting the known effect of Kirschmann's third law. These results indicate that Kirschmann's third law fails to explain the perceived color shift of our illusion, even after accounting for optical factors like aberrations. Observed color shifts could be explained by an augmented synergy theory based on intensity space, rather than chromaticity.

The Effect of Material Properties on the Perceived Shape of Three-Dimensional Objects.

Perceiving the shape of three-dimensional objects is essential for interacting with them in daily life. If objects are constructed from different materials, can the human visual system accurately estimate their three-dimensional shape? We varied the thickness, motion, opacity, and specularity of globally convex objects rendered in a photorealistic environment. These objects were presented under either dynamic or static viewing condition. Observers rated the overall convexity of these objects along the depth axis. Our results show that observers perceived solid transparent objects as flatter than the same objects rendered with opaque reflectance properties. Regional variation in local root-mean-square image contrast was shown to provide information that is predictive of perceived surface convexity.

Color Perception in Protanomalous Female Macaca fascicularis.

Protanomalous females with X chromosome-linked color vision deficiency exhibit mild abnormalities, whereas dichromats show a distinct deficiency in discriminating certain color pairs. Dichromats have an advantage in detecting a textured target when it is camouflaged by red-green colors, owing to their insensitivity to these colors. However, it is not certain whether protanomalous females possess a similar advantage in breaking camouflage. Here, we introduce an animal model of dichromatic macaque monkeys and protanomalous females. We examined whether protanomalous females have the same advantage in breaking color camouflage as shown by dichromatic macaques. We also tested whether they could discriminate a certain color pair that trichromats could, where the dichromats are confused. Our experiments show that protanomalous macaques can break color camouflage, similar to dichromats, and can discriminate colors similarly to trichromats. Protanomalous females are thus thought to have the combined ecological advantages of being both trichromats and dichromats.

Donut-Shaped Stretchable Kirigami: Enabling Electronics to Integrate with the Deformable Muscle.

Electronic devices used to record biological signals are important in neuroscience, brain-machine interfaces, and medical applications. Placing electronic devices below the skin surface and recording the muscle offers accurate and robust electromyography (EMG) recordings. The device stretchability and flexibility must be similar to the tissues to achieve an intimate integration of the electronic device with the biological tissues. However, conventional elastomer-based EMG electrodes have a Young's modulus that is ≈20 times higher than that of muscle. In addition, these stretchable devices also have an issue of displacement on the tissue surface, thereby causing some challenges during accurate and robust EMG signal recordings. In general, devices with kirigami design solve the issue of the high Young's modulus of conventional EMG devices. In this study, donut-shaped kirigami bioprobes are proposed to reduce the device displacement on the muscle surface. The fabricated devices are tested on an expanding balloon and they show no significant device (microelectrode) displacement. As the package, the fabricated device is embedded in a dissolvable material-based scaffold for easy-to-use stretchable kirigami device in an animal experiment. Finally, the EMG signal recording capability and stability using the fabricated kirigami device is confirmed in in vivo experiments without significant device displacements.

A Magnetically Assembled High-Aspect-Ratio Needle Electrode for Recording Neuronal Activity.

Microelectrode devices, which enable the detection of neuronal signals in brain tissues, have made significant contributions in the field of neuroscience and the brain-machine interfaces. To further develop such microelectrode devices, the following requirements must be met: i) a fine needle's diameter (<30 µm) to reduce damage to tissues; ii) a long needle (e.g., ≈1 mm for rodents and ≈2 mm for macaques); and iii) multiple electrodes to achieve high spatial recording (<100 µm in pitch). In order to meet these requirements, this study herein reports an assembly technique for high-aspect-ratio microneedles, which employs a magnet. The assembly is demonstrated, in which nickel wires of length 750 µm and diameter 25 µm are produced on a silicon substrate. The impedance magnitude of the assembled needle-like electrode measured at 1 kHz is 5.6 kΩ, exhibiting output and input signal amplitudes of 96.7% at 1 kHz. To confirm the recording capability of the fabricated device, neuronal signal recordings are performed using mouse cerebra in vivo. The packaged single microneedle electrode penetrates the barrel field in the primary somatosensory cortex of the mouse and enables the detection of evoked neuronal activity of both local field potentials and action potentials.

Fiber bundle endomicroscopy with multi-illumination for three-dimensional reflectance image reconstruction.

Bundled fiber optics allow in vivo imaging at deep sites in a body. The intrinsic optical contrast detects detailed structures in blood vessels and organs. We developed a bundled-fiber-coupled endomicroscope, enabling stereoscopic three-dimensional (3-D) reflectance imaging with a multipositional illumination scheme. Two illumination sites were attached to obtain reflectance images with left and right illumination. Depth was estimated by the horizontal disparity between the two images under alternative illuminations and was calibrated by the targets with known depths. This depth reconstruction was applied to an animal model to obtain the 3-D structure of blood vessels of the cerebral cortex (Cereb cortex) and preputial gland (Pre gla). The 3-D endomicroscope could be instrumental to microlevel reflectance imaging, improving the precision in subjective depth perception, spatial orientation, and identification of anatomical structures.

Ultrastretchable Kirigami Bioprobes.

An ultrastretchable film device is developed that can follow the shape of spherical and large deformable biological samples such as heart and brain tissues. Although the film is composed of biocompatible parylene for the device substrate and metal layers of platinum (Pt)/titanium (Ti), which are unstretchable materials, the film shows a high stretchability by patterning slits as a "Kirigami" design. A Pt/Ti-microelectrode array embedded in 11 µm thick parylene film with 5 × 91 slits exhibits a film strain of ≈250% at 9 mN strain-force (0.08 MPa in stress) with a Young's modulus of 23 kPa, while the 3 × 91-slit film shows a Young's modulus of 3.6 kPa. The maximum strains of these devices are ≈470% and ≈840%, respectively. It is demonstrated that the Kirigami-based microelectrode device can simultaneously record in vivo electrocorticogram signals from the visual and barrel cortices of a mouse by stretching the film and tuning the electrode gap. Moreover, wrapping the Kirigami device around a beating mouse's heart, which shows large and rapid changes in the volume and the surface area, can record the in vivo epicardial electrocardiogram signals. Such a small Young's modulus for a stretchable device reduces the device's strain-force, minimizing the device-induced stress to soft biological tissues.