Research articles and laboratory protocol organization often lack detailed instructions for replicating experiments. protocols.io is an open-access platform where researchers collaboratively create dynamic, interactive, step-by-step protocols that can be executed on mobile devices or the web. Researchers can easily and efficiently share protocols with colleagues, collaborators, the scientific community, or make them public. Real-time communication and interaction keep protocols up to date. Public protocols receive a DOI and enable open communication with authors and researchers to foster efficient experimentation and reproducibility.

Neuroplasticity is essential for the establishment and strengthening of neural circuits. Repetitive transcranial magnetic stimulation (rTMS) is commonly used to modulate cortical excitability and shows promise in the treatment of some neurological disorders. Low intensity magnetic stimulation (LI-rTMS), which does not directly elicit action potentials in the stimulated neurons, have also shown some therapeutic effects, and it is important to determine the biological mechanisms underlying the effects of these low intensity magnetic fields, such as would occur in the regions surrounding the central high-intensity focus of rTMS. Our team has used a focal low-intensity (10mT) magnetic stimulation approach to address some of these questions and to identify cellular mechanisms. I will present several studies from our laboratory, addressing (1) effects of LIrTMS on neuronal activity and excitability ; and (2) neuronal morphology and post-lesion repair. The ensemble of our results indicate that the effects of LI-rTMS depend upon the stimulation pattern, the age of the animal, and the presence of cellular magnetoreceptors.

Explore how microfluidic chips can enhance your imaging experiments by increasing control, throughput, or flexibility. In this remote, personalized workshop, participants will receive expert guidance, support and chips to run tests on their own microscopes.

Nervous system function relies on the polarized architecture of neurons, established by directional transport of pre- and postsynaptic cargoes. While delivery of postsynaptic components depends on the secretory pathway, the identity of the membrane compartment(s) that supply presynaptic active zone (AZ) and synaptic vesicle (SV) proteins is largely unknown. I will discuss our recent advances in our understanding of how key components of the presynaptic machinery for neurotransmitter release are transported and assembled focussing on our studies in genome-engineered human induced pluripotent stem cell-derived neurons. Specifically, I will focus on the composition and cell biological identity of the axonal transport vesicles that shuttle key components of neurotransmission to nascent synapses and on machinery for axonal transport and its control by signaling lipids. Our studies identify a crucial mechanism mediating the delivery of SV and active zone proteins to developing synapses and reveal connections to neurological disorders. In the second part of my talk, I will discuss how exocytosis and endocytosis are coupled to maintain presynaptic membrane homeostasis. I will present unpublished data regarding the role of membrane tension in the coupling of exocytosis and endocytosis at synapses. We have identified an endocytic BAR domain protein that is capable of sensing alterations in membrane tension caused by the exocytotic fusion of SVs to initiate compensatory endocytosis to restore plasma membrane area. Interference with this mechanism results in defects in the coupling of presynaptic exocytosis and SV recycling at human synapses.

Advancements in imaging speed, depth and resolution have made structured illumination microscopy (SIM) an increasingly powerful optical sectioning (OS) and super-resolution (SR) technique, but these developments remain inaccessible to many life science researchers due to the cost, optical complexity and delicacy of these instruments. We address these limitations by redesigning the optical path using in-line fibre components that are compact, lightweight and easily assembled in a “Plug & Play” modality, without compromising imaging performance. They can be integrated into an existing widefield microscope with a minimum of optical components and alignment, making OS-SIM more accessible to researchers with less optics experience. We also demonstrate a complete SR-SIM imaging system with dimensions 300 mm × 300 mm × 450 mm. We propose to enable accessible SIM imaging by utilising its compact, lightweight and robust design to transport it where it is needed, and image in “unconventional” environments where factors such as temperature and biosafety considerations currently limit imaging experiments.

OpenSPM aims to democratize innovation in the field of scanning probe microscopy (SPM), which is currently dominated by a few proprietary, closed systems that limit user-driven development. Our platform includes a high-speed OpenAFM head and base optimized for small cantilevers, an OpenAFM controller, a high-voltage amplifier, and interfaces compatible with several commercial AFM systems such as the Bruker Multimode, Nanosurf DriveAFM, Witec Alpha SNOM, Zeiss FIB-SEM XB550, and Nenovision Litescope. We have created a fully documented and community-driven OpenSPM platform, with training resources and sourcing information, which has already enabled the construction of more than 15 systems outside our lab. The controller is integrated with open-source tools like Gwyddion, HDF5, and Pycroscopy. We have also engaged external companies, two of which are integrating our controller into their products or interfaces. We see growing interest in applying parts of the OpenSPM platform to related techniques such as correlated microscopy, nanoindentation, and scanning electron/confocal microscopy. To support this, we are developing more generic and modular software, alongside a structured development workflow. A key feature of the OpenSPM system is its Python-based API, which makes the platform fully scriptable and ideal for AI and machine learning applications. This enables, for instance, automatic control and optimization of PID parameters, setpoints, and experiment workflows. With a growing contributor base and industry involvement, OpenSPM is well positioned to become a global, open platform for next-generation SPM innovation.

Sleep is an active state critical for processing emotional memories encoded during waking in both humans and animals. There is a remarkable overlap between the brain structures and circuits active during sleep, particularly rapid eye-movement (REM) sleep, and the those encoding emotions. Accordingly, disruptions in sleep quality or quantity, including REM sleep, are often associated with, and precede the onset of, nearly all affective psychiatric and mood disorders. In this context, a major biomedical challenge is to better understand the underlying mechanisms of the relationship between (REM) sleep and emotion encoding to improve treatments for mental health. This lecture will summarize our investigation of the cellular and circuit mechanisms underlying sleep architecture, sleep oscillations, and local brain dynamics across sleep-wake states using electrophysiological recordings combined with single-cell calcium imaging or optogenetics. The presentation will detail the discovery of a 'somato-dendritic decoupling'in prefrontal cortex pyramidal neurons underlying REM sleep-dependent stabilization of optimal emotional memory traces. This decoupling reflects a tonic inhibition at the somas of pyramidal cells, occurring simultaneously with a selective disinhibition of their dendritic arbors selectively during REM sleep. Recent findings on REM sleep-dependent subcortical inputs and neuromodulation of this decoupling will be discussed in the context of synaptic plasticity and the optimization of emotional responses in the maintenance of mental health.

What’s the point of having scientific and technological innovations when only a few can benefit from them? How can we make science more inclusive? Those questions are always in the back of my mind when we perform research in our laboratory, and we have a strong focus on the scientific accessibility of our developed methods from microfabrication to sensor development.

Astrocytes release glutamate by regulated exocytosis in health and disease Vladimir Parpura, International Translational Neuroscience Research Institute, Zhejiang Chinese Medical University, Hangzhou, P.R. China Parpura will present you with the evidence that astrocytes, a subtype of glial cells in the brain, can exocytotically release the neurotransmitter glutamate and how this release is regulated. Spatiotemporal characteristic of vesicular fusion that underlie glutamate release in astrocytes will be discussed. He will also present data on a translational project in which this release pathway can be targeted for the treatment of glioblastoma, the deadliest brain cancer.

A hallmark pathological feature of the neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) is the depletion of RNA-binding protein TDP-43 from the nucleus of neurons in the brain and spinal cord. A major function of TDP-43 is as a repressor of cryptic exon inclusion during RNA splicing. By re-analyzing RNA-sequencing datasets from human FTD/ALS brains, we discovered dozens of novel cryptic splicing events in important neuronal genes. Single nucleotide polymorphisms in UNC13A are among the strongest hits associated with FTD and ALS in human genome-wide association studies, but how those variants increase risk for disease is unknown. We discovered that TDP-43 represses a cryptic exon-splicing event in UNC13A. Loss of TDP-43 from the nucleus in human brain, neuronal cell lines and motor neurons derived from induced pluripotent stem cells resulted in the inclusion of a cryptic exon in UNC13A mRNA and reduced UNC13A protein expression. The top variants associated with FTD or ALS risk in humans are located in the intron harboring the cryptic exon, and we show that they increase UNC13A cryptic exon splicing in the face of TDP-43 dysfunction. Together, our data provide a direct functional link between one of the strongest genetic risk factors for FTD and ALS (UNC13A genetic variants), and loss of TDP-43 function. Recent analyses have revealed even further changes in TDP-43 target genes, including widespread changes in alternative polyadenylation, impacting expression of disease-relevant genes (e.g., ELP1, NEFL, and TMEM106B) and providing evidence that alternative polyadenylation is a new facet of TDP-43 pathology.

Visual restoration after decades of blindness is now becoming possible by means of retinal and cortical prostheses, as well as emerging stem cell and gene therapeutic approaches. After restoring visual perception, however, a key question remains. Are there optimal means and methods for retraining the visual cortex to process visual inputs, and for learning or relearning to “see”? Up to this point, it has been largely assumed that if the sensory loss is visual, then the rehabilitation focus should also be primarily visual. However, the other senses play a key role in visual rehabilitation due to the plastic repurposing of visual cortex during blindness by audition and somatosensation, and also to the reintegration of restored vision with the other senses. I will present multisensory neuroimaging results, cortical thickness changes, as well as behavioral outcomes for patients with Retinitis Pigmentosa (RP), which causes blindness by destroying photoreceptors in the retina. These patients have had their vision partially restored by the implantation of a retinal prosthesis, which electrically stimulates still viable retinal ganglion cells in the eye. Our multisensory and structural neuroimaging and behavioral results suggest a new, holistic concept of visual rehabilitation that leverages rather than neglects audition, somatosensation, and other sensory modalities.

High-quality glass lenses are commonplace in the design of optical instrumentation used across the biosciences. However, research-grade glass lenses are often costly, delicate and, depending on the prescription, can involve intricate and lengthy manufacturing - even more so in bioimaging applications. This seminar will outline 3D printing as a viable low-cost alternative for the manufacture of high-performance optical elements, where I will also discuss the creation of the world’s first fully 3D printed microscope and other implementations of 3D printed lenses. Our 3D printed lenses were generated using consumer-grade 3D printers and pose a 225x materials cost-saving compared to glass optics. Moreover, they can be produced in any lab or home environment and offer great potential for education and outreach. Following performance validation, our 3D printed optics were implemented in the production of a fully 3D printed microscope and demonstrated in histological imaging applications. We also applied low-cost fabrication methods to exotic lens geometries to enhance resolution and contrast across spatial scales and reveal new biological structures. Across these applications, our findings showed that 3D printed lenses are a viable substitute for commercial glass lenses, with the advantage of being relatively low-cost, accessible, and suitable for use in optical instruments. Combining 3D printed lenses with open-source 3D printed microscope chassis designs opens the doors for low-cost applications for rapid prototyping, low-resource field diagnostics, and the creation of cheap educational tools.

Fast periodic visual stimulation (FPVS) has emerged as a promising tool for assessing cognitive function in individuals with dementia. This technique leverages electroencephalography (EEG) to measure brain responses to rapidly presented visual stimuli, offering a non-invasive and objective method for evaluating a range of cognitive functions. Unlike traditional cognitive assessments, FPVS does not rely on behavioural responses, making it particularly suitable for individuals with cognitive impairment. In this talk I will highlight a series of studies that have demonstrated its ability to detect subtle deficits in recognition memory, visual processing and attention in dementia patients using EEG in the lab, at home and in clinic. The method is quick, cost-effective, and scalable, utilizing widely available EEG technology. FPVS holds significant potential as a functional biomarker for early diagnosis and monitoring of dementia, paving the way for timely interventions and improved patient outcomes.

Pharmacokinetics in the eye is an important factor for the success of ocular drug delivery and treatment. Pharmacokinetic features determine the feasible routes of drug administration, dosing levels and intervals, and it has impact on eventual drug responses. Several physical, biochemical, and flow-related barriers limit drug exposure of anterior and posterior ocular target tissues during treatment during local (topical, subconjunctival, intravitreal) and systemic administration (intravenous, per oral). Mathematical models integrate joint impact of various barriers on ocular pharmacokinetics (PKs) thereby helping drug development. The models are useful in describing (top-down) and predicting (bottom-up) pharmacokinetics of ocular drugs. This is useful also in the design and development of new drug molecules and drug delivery systems. Furthermore, the models can be used for interspecies translation and probing of disease effects on pharmacokinetics. In this lecture, ocular pharmacokinetics and current modelling methods (noncompartmental analyses, compartmental, physiologically based, and finite element models) are introduced. Future challenges are also highlighted (e.g. intra-tissue distribution, prediction of drug responses, active transport).

Overview of neurosurgery specialty interplay between neurology, psychiatry and neurosurgery. Discussion on benefits and disadvantages of classifications. Presentation of sub-specialties: trauma, oncology, functional, pediatric, vascular and spine. How does an ordinary day of a neurosurgeon look like; outpatient clinic, emergencies, pre/intra/post operative patient care. An ordinary operation. Myth-busting and practical insights of every day practice. An ordinary operation. Hint for research on clinical problems to be solved. The coming ethical frontiers of neuroprosthetics. In part two we will explore the explanatory gap and its significance. We will review the more than 200 theories of the hard problem of consciousness, from the prevailing to the unconventional. Finally, we are going to reflect on the AI advancements and the claims of LLMs becoming conscious

Photoreceptor loss is the primary cause behind vision impairment and blindness in diseases such as retinitis pigmentosa and age-related macular degeneration. However, the death of rods and cones allows retinoids to permeate the inner retina, causing retinal ganglion cells to become spontaneously hyperactive, severely reducing the signal-to-noise ratio, and creating interference in the communication between the surviving retina and the brain. Treatments aimed at blocking or reducing hyperactivity improve vision initiated from surviving photoreceptors and could enhance the signal fidelity generated by vision restoration methodologies.

Unlike traditional monoamine-based antidepressants that require weeks to exert effects, ketamine alleviates depression within hours, though its clinical use is limited by side effects. While ketamine was initially thought to work primarily through NMDA receptor (NMDAR) inhibition, our research reveals a more complex mechanism. We demonstrate that NMDAR inhibition alone cannot explain ketamine's sustained antidepressant effects, as other NMDAR antagonists like MK-801 lack similar efficacy. Instead, the (2R,6R)-hydroxynorketamine (HNK) metabolite appears critical, exhibiting antidepressant effects without ketamine's side effects. Paradoxically, our findings suggest an inverted U-shaped dose-response relationship where excessive NMDAR inhibition may actually impede antidepressant efficacy, while some level of NMDAR activation is necessary. The antidepressant actions of ketamine and (2R,6R)-HNK require AMPA receptor activation, leading to synaptic potentiation and upregulation of AMPA receptor subunits GluA1 and GluA2. Furthermore, NMDAR subunit GluN2A appears necessary and possibly sufficient for these effects. This research establishes NMDAR-GluN2A activation as a common downstream effector for rapid-acting antidepressants, regardless of their initial targets, offering promising directions for developing next-generation antidepressants with improved efficacy and reduced side effects.

La resonancia magnética nuclear está basada en el fenómeno del magnetismo nuclear que más aplicaciones ha encontrado para el estudio de enfermedades humanas. Usualmente la señal de RM es recibida y transmitida a distancias cercanas al objeto del que se quiere obtener una imagen. Otra alternativa es emitir y recibir la misma señal de manera remota haciendo uso de guías de onda. Este enfoque tiene la ventaja que se puede aplicar a altos campos magnéticos, la absorción de energía es menor, además es posible cubrir mayores regiones de interés y comodidad para el paciente. Por otro lado, sufre de baja calidad de imagen en algunos casos. En esta ocasión hablaremos de nuestra experiencia haciendo uso de este enfoque empleando una guía de ondas abierta y metamateriales tanto para sistemas clínicos como preclínicos de IRM.

Neurotrophins (NGF, BDNF, NT-3) are endogenous growth factors that exert neuroprotective effects by preventing neuronal death and promoting neurogenesis. They act by binding to their respective high-affinity, pro-survival receptors TrkA, TrkB or TrkC, as well as to p75NTR death receptor. While these molecules have been shown to significantly slow or prevent neurodegeneration, their reduced bioavailability and inability to penetrate the blood-brain-barrier limit their use as potential therapeutics. To bypass these limitations, our research team has developed and patented small-sized, lipophilic compounds which selectively resemble neurotrophins’ effects, presenting preferable pharmacological properties and promoting neuroprotection and repair against neurodegeneration. In addition, the combination of these molecules with 3D cultured human neuronal cells, and their targeted delivery in the brain ventricles through soft robotic systems, could offer novel therapeutic approaches against neurodegenerative diseases and brain trauma.

Do You Know Your Blood Glucose Level? You Probably Should! A single measurement is not enough to truly understand your metabolic health. Blood glucose levels fluctuate dynamically, and meaningful insights require continuous monitoring over time. But glucose is just one example. Many other molecular concentrations in the body are not static. Their variations are influenced by individual physiology and overall health. PhenoSign, a Swiss MedTech startup, is on a mission to become the leader in real-time molecular analysis of complex fluids, supporting clinical decision-making and life sciences applications. By providing real-time, in-situ molecular insights, we aim to advance medicine and transform life sciences research. This talk will provide an overview of PhenoSign’s journey since its inception in 2022—our achievements, challenges, and the strategic roadmap we are executing to shape the future of real-time molecular diagnostics.