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The rise of microscopy in the seventeenth century allowed scientists to discover a new world of microorganisms and achieve great physiological advances. One of the first microscopes of the epoch was Antonie van Leeuwenhoek’s microscope, a deceptively simple device that contains a single ball lens housed in a metal plate allowing the observation of samples at up to ×250 magnification. Such magnification was much greater than that achieved by rudimentary compound microscopes of the era, allowing for the discovery of microscopic, single-celled life, an achievement that marked the study of biology up to the nineteenth century. Since Leeuwenhoek’s design uses a single ball lens, it is possible to fabricate variations for educational activities in physics and biology university and high school classrooms. A fundamental problem, however, with home-built microscopes is that it is difficult to work with glass. We developed a simple protocol to make ball lenses of glass and gelatin with high magnification that can be done in a university/high school classroom, and we designed an optimized support for focusing and taking photographs with a smartphone. The protocol details a simple, easily accessible, low-cost, and effective tool for the observation of microscopic samples, possible to perform anywhere without the need for a laboratory or complex tools. Our protocol has been implemented in classrooms in Chile to a favorable reception.

Development of advanced chronic, high-fidelity neural interfaces is accelerating research into brain function[1] and more effective treatments for neurological conditions.[2–5] The primary functional requirements of these interfaces include recording and/or stimulating from a number of discretely sampled volumes of the brain at requisite spatial resolutions for specific time periods that may extend from hours to years.[6–9] This translates to a push towards smaller electrodes that are more biologically transparent and biocompatible[10–12] with a high density of electrode sites that remain functional for long period of time.[13–15] As electrode size goes to the microscale (higher spatial selectivity), the impedance of electrode site increases, and consequently, the quality of signal recordings decreases (lower sensitivity). Thus, there is a trade off between the size (spatial selectivity) and quality of signal recordings (sensitivity) in neural microelectrodes.[16–18] Studies have also shown that the response of brain tissue to implanted microelectrodes includes an acute injury and a chronic reactive tissue response. The chronic response is characterized by the presence of both activated microglia and reactive astrocytes, which eventually encapsulate the electrode to some degree.[10,12,19–21] Consequently, in addition to the initial high impedance of microelectrodes, these reactive tissue responses have been associated with a progressive increase in the impedance of the electrode/tissue interface.[10,12] Therefore, achieving a very low impedance electrode/tissue interface is important for maintaining and improving signal quality.

One of the great unsolved problems in science is deciphering the neural code for long-term memory or how the brain stores and represents information. Simply put: How are we that we are? How is longterm memory stored? How is it encoded? How is it lost? Can we hope to build a machine to do what we do naturally? This is an introduction to the both ancient and contemporary problem of the neural code and we explore a series of thought experiments examining how the brain encodes longterm memory and possible theoretical storage limits.

The introduction of affordable, consumer-oriented 3-D printers is a milestone in the current "maker movement," which has been heralded as the next industrial revolution. Combined with free and open sharing of detailed design blueprints and accessible development tools, rapid prototypes of complex products can now be assembled in one's own garage--a game-changer reminiscent of the early days of personal computing. At the same time, 3-D printing has also allowed the scientific and engineering community to build the "little things" that help a lab get up and running much faster and easier than ever before.

Recent neuroprosthetic work has focused on the motor cortex as a source of voluntary control signals. However, the motor cortex can be damaged in upper motor neuron degenerative diseases such as primary lateral sclerosis and amyotrophic lateral sclerosis. The possibility exists that prefrontal areas may also be used in neuroprosthetic devices. Here, we report the use of the cingulate cortex in a neuroprosthetic model. Seven rats were able to significantly modulate spiking activity in the cingulate cortex in order to receive reward. Furthermore, experiments with single neurons provide evidence that the cingulate cortex neuronal modulation is highly flexible and thus useful for a neuroprosthetic device.

Mars is a compelling astrobiological target, and a human mission would provide an opportunity to collect immense amounts of scientific data. Exploration alone, however, cannot justify the increased risk. Instead, three factors drive a human mission: economics, education, and exploration. A human mission has a unique potential to inspire the next generation of young people to enter critically needed science and engineering disciplines. A mission is economically feasible, and the research and development program put in place for a human mission would propel growth in related high-technology industries. The main hurdles are human physiological responses to 1-2 years of radiation and microgravity exposure. However, enabling technologies are sufficiently mature in these areas that they can be developed within a few decade timescale. Hence, the decision of whether or not to undertake a human mission to Mars is a political decision, and thus, educational and economic benefits are the crucial factors.

Severed segments of rat peripheral (PNS; sciatic) and central nervous system (CNS; spinal) axons continue to conduct action potentials when maintained in vitro at 6-9 degrees C for up to 7 (sciatic axons) and 2 days (spinal axons), compared with only 36 h at 37-38 degrees C for sciatic axons and 6 h for spinal axons. These PNS and CNS axonal segments can be crushed and then treated with polyethylene glycol (PEG), resulting in a rapid reconnection (fusion) of the surviving axons at the crush site, as assessed by conduction of action potentials through the crush site within minutes after PEG administration. Severed PNS or CNS axons maintained in vitro at 6-9 degrees C prior to crushing can be successfully PEG-fused for up to 4 and 1.5 days, respectively, compared with only 24 (sciatic) and 3 h (spinal) at 37-38 degrees C. These data demonstrate that cooling significantly increases both the survival time of severed mammalian PNS and CNS axons and the time that severed axons can still be PEG-fused (rejoined) to rapidly re-establish axonal continuity in vitro.

The advent of easy-to-use open source microcontrollers, off-the-shelf electronics and customizable manufacturing technologies has facilitated the development of inexpensive scientific devices and laboratory equipment. In this study, we describe an imaging system that integrates low-cost and open-source hardware, software and genetic resources. The multi-fluorescence imaging system consists of readily available 470 nm LEDs, a Raspberry Pi camera and a set of filters made with low cost acrylics. This device allows imaging in scales ranging from single colonies to entire plates. We developed a set of genetic components (e.g. promoters, coding sequences, terminators) and vectors following the standard framework of Golden Gate, which allowed the fabrication of genetic constructs in a combinatorial, low cost and robust manner. In order to provide simultaneous imaging of multiple wavelength signals, we screened a series of long stokes shift fluorescent proteins that could be combined with cyan/green fluorescent proteins. We found CyOFP1, mBeRFP and sfGFP to be the most compatible set for 3-channel fluorescent imaging. We developed open source Python code to operate the hardware to run time-lapse experiments with automated control of illumination and camera and a Python module to analyze data and extract meaningful biological information. To demonstrate the potential application of this integral system, we tested its performance on a diverse range of imaging assays often used in disciplines such as microbial ecology, microbiology and synthetic biology. We also assessed its potential for STEM teaching in a high school environment, using it to teach biology, hardware design, optics, and programming. Together, these results demonstrate the successful integration of open source hardware, software, genetic resources and customizable manufacturing to obtain a powerful, low cost and robust system for STEM education, scientific research and bioengineering. All the resources developed here are available under open source licenses.

The earthworm is ideal for studying action potential conduction velocity in a classroom setting, as its simple linear anatomy allows easy axon length measurements and the worm's sparse coding allows single action potentials to be easily identified. The earthworm has two giant fiber systems (lateral and medial) with different conduction velocities that can be easily measured by manipulating electrode placement and the tactile stimulus. Here, we present a portable and robust experimental setup that allows students to perform conduction velocity measurements within a 30-min to 1-h laboratory session. Our improvement over this well-known preparation is the combination of behaviorally relevant tactile stimuli (avoiding electrical stimulation) with the invention of minimal, low-cost, and portable equipment. We tested these experiments during workshops in both a high school and college classroom environment and found positive learning outcomes when we compared pre- and posttests taken by the students.

While the development of microelectrode arrays has enabled access to disparate regions of a cortex for neurorehabilitation, neuroprosthetic and basic neuroscience research, accurate interpretation of the signals and manipulation of the cortical neurons depend upon the anatomical placement of the electrode arrays in a layered cortex. Toward this end, this report compares two in vivo methods for identifying the placement of electrodes in a linear array spaced 100 µm apart based on in situ laminar analysis of (1) ketamine-xylazine-induced field potential oscillations in a rat motor cortex and (2) an intracortical electrical stimulation-induced movement threshold. The first method is based on finding the polarity reversal in laminar oscillations which is reported to appear at the transition between layers IV and V in laminar 'high voltage spindles' of the rat cortical column. Analysis of histological images in our dataset indicates that polarity reversal is detected 150.1 ± 104.2 µm below the start of layer V. The second method compares the intracortical microstimulation currents that elicit a physical movement for anodic versus cathodic stimulation. It is based on the hypothesis that neural elements perpendicular to the electrode surface are preferentially excited by anodic stimulation while cathodic stimulation excites those with a direction component parallel to its surface. With this method, we expect to see a change in the stimulation currents that elicits a movement at the beginning of layer V when comparing anodic versus cathodic stimulation as the upper cortical layers contain neuronal structures that are primarily parallel to the cortical surface and lower layers contain structures that are primarily perpendicular. Using this method, there was a 78.7 ± 68 µm offset in the estimate of the depth of the start of layer V. The polarity reversal method estimates the beginning of layer V within ±90 µm with 95% confidence and the intracortical stimulation method estimates it within ±69.3 µm. We propose that these methods can be used to estimate the in situ location of laminar electrodes implanted in the rat motor cortex.

Closed-loop neural interface technology that combines neural ensemble decoding with simultaneous electrical microstimulation feedback is hypothesized to improve deep brain stimulation techniques, neuromotor prosthetic applications, and epilepsy treatment. Here we describe our iterative results in a rat model of a sensory and motor neurophysiological feedback control system. Three rats were chronically implanted with microelectrode arrays in both the motor and visual cortices. The rats were subsequently trained over a period of weeks to modulate their motor cortex ensemble unit activity upon delivery of intra-cortical microstimulation (ICMS) of the visual cortex in order to receive a food reward. Rats were given continuous feedback via visual cortex ICMS during the response periods that was representative of the motor cortex ensemble dynamics. Analysis revealed that the feedback provided the animals with indicators of the behavioral trials. At the hardware level, this preparation provides a tractable test model for improving the technology of closed-loop neural devices.

A small (1cm(3)), light-weight (1g including batteries), low power (10mW, lasts 25 hrs), long range (22 ft.), 3-channel wireless neural recording system is designed, fabricated and characterized through in-vitro and in-vivo experiments. The transmitter comprises of an ASIC fabricated in 2-Poly, 2-Metal 1.5 microm AMIS process which can transmit data out wirelessly using on-chip VCO or low power commercial off-the-shelf (COTS) transmitters. The microsystem is employed in collecting neural signals from two different animal models: axons in cockroach leg and forelimb area of the motor cortex of a mature Long Evans rat.

Local field potentials (LFPs) have been proposed for use in controlling neural prosthetic devices because they can provide reliable motor and sensory-related information, and can easily be recorded over long periods of time. While studies have shown that directional information about motor movements can be inferred from LFPs, it is not known at what depth these signals should be recorded from in order to maximize the amount of movement information. Towards this end, we used a directional motor task in Long Evans rats, while sampling LFPs with an electrode consisting of 16 vertical recording sites that were evenly-spaced 100μm apart. This allowed for simultaneous recording of all layers of the motor cortex. The frequency components of LFPs were then analyzed using k-means clustering to determine directional information as a function of depth. Here we report our initial findings that superficial layers (II/III) of motor cortex may provide more information about movement directions then deeper layers (V). Information analysis of LFPs was conducted using a non- parametric clustering algorithm that estimated the statistical similarity of LFP frequency components associated with leftward and rightward movements using data from all layers of the rat primary motor cortex. Our findings suggest that the superficial layers of the motor cortex may be optimal for determining the movement direction from LFPs. We also provide evidence, using current source density (CSD) analysis, that the better decoding may be due to the synaptic activity in the upper layers.

This paper presents a review of our neural interface research program and provides a brief introduction to the field. We focus on three key areas which aim to minimize encapsulating tissue response to chronic neural interfaces: interface architecture, mechanical stresses, and stem cell-seeded probes. We explore these areas and present our current solutions which have led to the demonstration of cortical control of neuromotor prostheses (NMPs).

While the development of silicon-substrate microelectrode arrays has enabled chronic recording of single unit activity from multiple neurons simultaneously, accurate interpretation of the signals depend on the anatomical placement of the electrodes. Toward this end, this paper develops an in vivo method for identifying the placement of electrodes based on laminar analysis of ketamine-xylazien-induced field potential oscillations in rat motor cortex. The proposed method is based on finding the polarity reversal in laminar oscillations which is reported to appear in upper part of layer V in laminar High Voltage Spindles (HVSs) of rat cortical column. Analysis of histological images showed a 21 mum error in the estimate of the polarity reversal depth compared to the expected range (850-1050 mum). One out of the four rats did not undergo a phase reversal. Histology verified that the electrode was placed deeper than 1050 jim. We propose that this method can be used to determine an estimate of laminar electrodes implanted in rat motor cortex

Developments of the past decades have shown it is possible to decode and extract control signals directly from the brain for use in neuromotor prosthetics. Currently, there is a push towards developing closed loop interface systems. Here we describe our early results in a rat model of a sensory/motor neurophysiological feedback control system. Traditional operant conditioning techniques historically train an animal to increase or decrease the occurrence of a behavior in response to a stimulus using various types of reinforcement. Here we demonstrate a rat performing an operant conditioning task with the modification that the rat's behavioral output and stimulus input are accessed via direct electrical connections with the brain. Our preliminary results here show a rat successfully trained to modulate the firing rates of its motor cortex in response to intra-cortical microstimulation of the visual cortex.

The interface between neural prostheses and neural tissue plays a significant role in the long term performance of these devices. Conducting polymers have been used to modify the electrical properties of neural microelectrodes. The objective of this study was to evaluate recording chronic neural activity of neural microelectrodes that were modified with nanofibers-templated of poly (3,4-ethylenedioxythiophene) (PEDOT) nanotubes over seven week periods using impedance spectroscopy and signal-to-noise ratio measurements. PEDOT nanotubes-coated sites were found to have lower impedance and higher signal-to-noise ratio than control site.

Brain machine interface development typically falls into two arenas, invasive extracellular recording and non-invasive electroencephalogram recording methods. The relationship between action potentials and field potentials is not well understood, and investigation of interrelationships may improve design of neuroprosthetic control systems. Rats were trained on a motor learning task whereby they had to insert their noses into an aperture while simultaneously pressing down on levers with their forepaws; spikes, local field potentials (LFPs), and electrocorticograms (ECoGs) over the motor cortex were recorded and characterized. Preliminary results suggest that the LFP activity in lower cortical layers oscillates with the ECoG.

Traditional brain-machine interfaces have typically focused on methods that use rate-based codes as a source for control signals. Opposed to rate, timing of firing across different neurons and within each neuron could also provide information that can be used for controlling brain-machine interfaces or neuroprosthetic devices. Findings have indicated that synchronization of individual spike discharges may help serve the organization of cortical motor processes. We are investigating neural firing synchrony in the context of using it for real-time control for neuroprostheses systems. Our results with rats suggest that subjects can be trained to synchronize neural firing and increase unitary events i.e. spike coincidence patterns that are significantly above chance. Temporal coding methods could be used as additional or alternative cortical control signals for neuroprostheses and brain machine interfaces.

Protein synthesis inhibitors (PSIs) increase the rate of degeneration, as measured by compound action potential (CAP) conduction, in segments of rat PNS and CNS axons. Sciatic axonal segments maintained in vitro in Krebs at 37-38 degrees C generate CAPs for 24 h compared to 8 h for axons exposed to Krebs containing two PSIs, 100 microM anisomycin and/or 35 microM cycloheximide. Spinal axonal segments at 37-38 degrees C generate CAPs for 3 h compared to 2 h for axons exposed to Krebs containing PSIs. While cooling (6-9 degrees C) slows degeneration rate, cooled sciatic axons exposed to PSIs exhibit lower peak CAPs compared to cooled control segments (P0.005).

Brain–machine interface decoding algorithms need to be predicated on assumptions that are easily met outside of an experimental setting to enable a practical clinical device. Given present technological limitations, there is a need for decoding algorithms which (a) are not dependent upon a large number of neurons for control, (b) are adaptable to alternative sources of neuronal input such as local field potentials (LFPs), and (c) require only marginal training data for daily calibrations. Moreover, practical algorithms must recognize when the user is not intending to generate a control output and eliminate poor training data. In this paper, we introduce and evaluate a Bayesian maximum-likelihood estimation strategy to address the issues of isolating quality training data and self-paced control. Six animal subjects demonstrate that a multiple state classification task, loosely based on the standard center-out task, can be accomplished with fewer than five engaged neurons while requiring less than ten trials for algorithm training. In addition, untrained animals quickly obtained accurate device control, utilizing LFPs as well as neurons in cingulate cortex, two non-traditional neural inputs.

Penetrating neural probe technologies allow investigators to record electrical signals in the brain. The implantation of probes causes acute tissue damage, partially due to vasculature disruption during probe implantation. This trauma can cause abnormal electrophysiological responses and temporary increases in neurotransmitter levels, and perpetuate chronic immune responses. A significant challenge for investigators is to examine neurovascular features below the surface of the brain in vivo. The objective of this study was to investigate localized bleeding resulting from inserting microscale neural probes into the cortex using two-photon microscopy (TPM) and to explore an approach to minimize blood vessel disruption through insertion methods and probe design. 3D TPM images of cortical neurovasculature were obtained from mice and used to select preferred insertion positions for probe insertion to reduce neurovasculature damage. There was an 82.8 +/- 14.3% reduction in neurovascular damage for probes inserted in regions devoid of major (5 microm) sub-surface vessels. Also, the deviation of surface vessels from the vector normal to the surface as a function of depth and vessel diameter was measured and characterized. 68% of the major vessels were found to deviate less than 49 microm from their surface origin up to a depth of 500 microm. Inserting probes more than 49 microm from major surface vessels can reduce the chances of severing major sub-surface neurovasculature without using TPM.

Improving cortical prostheses requires the development of recording neural interfaces that are efficient in terms of providing maximal control information with minimal interface complexity. While the typical approaches have targeted neurons in the motor cortex with multiple penetrating shanks, an alternative approach is to determine an efficient distribution of electrode sites within the layers of the cortex with fewer penetrating shanks. The objective of this study was to compare unit activity in the upper and lower layers of the cortex with respect to movement and direction in order to inform the design of penetrating microelectrodes. Four rats were implanted bilaterally with multi-site single-shank silicon microelectrode arrays in the neck/shoulder region of the motor cortex. We simultaneously recorded unit activity across all layers of the motor cortex while the animal was engaged in a movement direction task. Localization of the electrode array within the different layers of the cortex was determined by histology. We denoted units from layers 2 and 3 and units as upper layer units, and units from layers 5 and 6 as lower layer units. Analysis of unit spiking activity demonstrated that both the upper and lower layers encode movement and direction information. Unit responses in either cortical layer of the cortex were not preferentially associated with contralateral or ipsilateral movement. Aggregate analysis (633 neurons) and best session analysis (75 neurons) indicated that units in the lower layers (layers 5, 6) are more likely to encode direction information when compared to units in the upper layers (layers 2, 3) (p< 0.05). These results suggest that electrode sites clustered in the lower layers provide access to more salient control information for cortical neuroprostheses.

The neocortex is a six-layered tissue consisting of different cell types. How does unit activity in the different layers of the motor cortex relate to movement? Does implantation in a particular layer improve direction decoding ability for a neuroprosthetic device? We simultaneously recorded unit activity in different layers of the rat motor cortex using chronic multi-site silicon electrodes. We used a combination of histology and electrophysiological signatures of Local Field Potentials (LFPs) to accurately localize the electrode sites in the different layers of the cortex. We analyzed 142 units from two animals and found that 40 units (28%) in Layers II to V showed significant modulation with respect to movement. Of these units that showed significant modulation, 9/20 (45%) of units in Layers II/III encoded directional information as compared to 15/19 (79%) of the units in Layers IV/V. These preliminary results suggest that units in Layers IV/V relatively contain more directional information than other layers of the cortex.