Global and community-driven initiatives have recently achieved considerable success in overcoming key challenges that hinder the widespread adoption of advanced microscopy and bioimage analysis tools in under-resourced settings. To build upon this progress, we held a workshop in May 2025 at the University of York, UK to address the needs and barriers associated with implementing time-lapse imaging and machine learning-based phenotyping in low-resource research environments. We focussed on identifying the specific challenges faced by the existing networks represented at the meeting, emphasising how integrating combined imaging hardware and machine learning-based approaches can solve these problems. This article summarises the key observations and actionable strategies made at the workshop. These proposed steps aim to significantly increase the dissemination and uptake of these powerful technologies to advance biological research in low-resource settings globally.

Emerging spatial multi-omics technologies enable the profiling of molecular variation within its tissue context, yet existing methods for identifying spatially variable features lack principled approaches to experimental design and cross-sample inference. Here, we present STORM, a principled Statistical TOol for spatially Resolved Multi-omics, for rigorously analyzing spatial patterns in spatial multi-omics research. STORM incorporates a robust and efficient nonparametric test that quantifies local deviations in molecular feature measurements to detect spatial dependence across transcriptomic and proteomic data. It further estimates an interpretable spatial effect size, supports power calculations for both spatial locations and biological replicates, and enables formal group-level comparisons. In several simulated and experimental spatial multi-omics case studies, STORM demonstrates reliable performance in detecting spatial structures while offering quantitative support for study design decisions. Overall, STORM provides a principled statistical framework that unifies spatial hypothesis testing, effect size estimation, power analysis, and experimental design for spatial multi-omics data.

Motor systems must flexibly select between competing outputs while preserving stability of rhythmic outputs. In Drosophila larvae, the isolated central nervous system is capable of maintaining rhythmicity by generating multiple different fictive motor programs. The biogenic amines octopamine and tyramine are known to regulate larval locomotion, however, how the tdc2+ octopaminergic/tyraminergic system regulates motor program competition is not well understood. Here, we combine dual-colour calcium imaging and optogenetic manipulation to explore how tdc2+ neurons track, permit, and bias fictive motor activity in 3rd instar Drosophila larvae. We find that tdc2+ activity in the larval ventral nerve cord is strongly coupled to motor neuron activity across multiple fictive behaviours, indicating that the system is recruited broadly across the motor repertoire. Optogenetic depolarisation of tdc2+ neurons increases motor root bursting and induces a robust fictive forward bias, whereas optogenetic hyperpolarisation suppresses or abolishes fictive rhythms and generates a short-lasting, post-inhibitory rebound in fictive activity. Spatially-restricted stimulation reveals that posterior abdominal activation is especially effective at promoting fictive forward activity. Separating VNC-residing from brain-residing tdc2+ populations further shows that activation of descending brain-residing tdc2+ projections is sufficient to recapitulate this forward bias. Finally, tdc2+ activation induces short-lived post-stimulation changes in motor programme probability, including transient elevation of competing fictive backward instantaneous frequency. Together, these findings suggest that tdc2+ neurons act as a permissive and biasing modulatory layer within larval motor circuits, linking adrenergic-like signalling to motor programme competition.

The cellular distribution of mitochondria in response to stress and local energy needs is governed by the relative activities of the microtubule-based molecular motors kinesin and dynein. The mechanism for switching between these two opposite-polarity microtubule motors remains unknown. In this study, we coupled a cellular synthetic cargo transport assay with AlphaFold2-guided mutagenesis to identify a regulatory helix in the mitochondrial adaptor protein [trafficking kinesin-binding protein (TRAK)] that mediates switching between kinesin- and dynein-driven transport. Differences in the helix sequence explained why two near-identical TRAK isoforms transported mitochondria in predominantly opposite directions. Phosphorylation of the regulatory helix by stress-activated kinases caused the activation of dynein and dissociation of kinesin. Our results reveal a molecular mechanism for coordinating the directional transport of mitochondria in response to intracellular signals.

Postural control, the capacity of an animal to detect and correct an inappropriate body orientation, is a widespread and fundamental neurobiological function, yet the neural circuits that implement it remain poorly resolved in most species. The larva of Drosophila melanogaster performs a posture control stereotyped self-righting (SR) manoeuvre when turned upside-down, a behaviour previously shown to depend on a pair of segmentally repeated lateral transverse motor neurons (LT1/2-MNs) and on the normal expression of several Hox-targeting microRNAs. Here, we use a connectomics and a functional behavioural approach to trace and test the sensory, interneuronal and motor architecture of the SR circuit along the antero-posterior axis. Starting from the LT1/2-MNs, we identify a set of pre-motor interneurons, their principal upstream partners, and a population of class IV multidendritic sensory neurons as core components of the circuit, and show, through neuron-specific thermogenetic silencing, that inhibiting the great majority of these elements significantly impairs SR performance. We further find substantial overlap between the SR circuit and the previously described nociceptive rolling and touch crawling circuits, converging on shared interneurons including DnB, A02o (Wave-1) and TePn05. Network and axial connectivity analyses reveal a nested, hub-like organisation, a small number of integrator neurons bridging sensory and motor sub-networks, and a consistent decline in synapse number and density towards posterior segments. Together, these data yield the first axial wiring diagram for a postural control circuit in any animal and offer a set of structural principles including: hub organisation, shared sensory-motor structure, and antero-posterior connectivity gradients, which may extend to other segmentally organised nervous systems.

The brain orchestrates billions of neurons spanning hundreds of regions to drive behavior. High-density electrophysiology has provided insights into this process, but monitoring brain-wide patterns of spiking activity demands new tools. Here we present Neuropixels 2.0 Quad Base, a 1536-channel probe with four times the simultaneous recording capacity of state-of-art Neuropixels 2.0. Using two Quad Base probes, we performed 3072-channel recordings in mice trained to produce complex sequences of goal-directed licks. We decoded behavioral kinematics from neural populations across more than 20 brain regions and identified the spatiotemporal emergence of motor decision signals. The probes' high channel count further revealed expanded inter-neuronal functional subnetworks and latent dynamics across multiple areas-features significantly undersampled when channels were subsampled to mimic standard Neuropixels 2.0. Neuropixels 2.0 Quad Base offers a powerful tool to explore neural encoding of cognition and behavior at grand scale.

Light microscopy is one of the fundamental readout methods in biomedical research, supporting scientific discovery across research fields, spatial scales, and model organisms [1,2]. Modern microscopy excels at molecular tagging [3], enabling visualization of biomolecules with growing sensitivity and resolution [4,5]; yet many foundational questions hinge on properties that cannot be easily labelled and quantified with traditional methods: the mechanical properties of biological samples [6], the organization of extracellular matrix in live organisms [7], and the spatiotemporal ion dynamics during neuronal activity [8]. This talk highlights three technology development that extend light microscopy capabilities to such readouts: (i) Brillouin microscopy for all-optical biophysical characterization, (ii) the glycan-binding fluorophore Rhobo6 for live extracellular matrix (ECM) imaging, and (iii) a novel toolkit of small molecule sensors to map extracellular calcium dynamics.

Small GTPases are generally viewed as organizers of organelle identity, with different families recruited to specific membranes by compartment-selective targeting mechanisms. Here, we identify a membrane state that reverses this relationship. Excessive peptide S-palmitoylation at Golgi generates multilamellar, filipin-poor whorls that recruit ARF, Rab, and Rho family GTPases normally associated with distinct cellular compartments. The whorls simultaneously excluded Golgi transmembrane residents, coat proteins, ER proteins, a GPI-anchored protein, and other palmitoylated proteins, demonstrating that they were selective membrane compartments rather than nonspecific protein aggregates. Preventing myristoylation of ARF6 or geranylgeranylation of Rab11a strongly reduced recruitment, whereas prenylation alone was insufficient, indicating recognition of a composite membrane-targeting code. Lowering cellular cholesterol promoted whorl formation, whereas cholesterol supplementation suppressed it. Chemically distinct lipid perturbations generated GTPase-positive Golgi whorls, suggesting convergence on a shared membrane-remodeling state. As whorls accumulated, endocytic recycling and anterograde trafficking declined, survival signaling decreased, and viability became poorly reversible. These findings show that lipid imbalance can redirect small GTPases across normal organelle boundaries, converting Golgi-derived membranes into selective sinks for trafficking regulators.

We introduce a workflow to identify oligomeric structures that are recorded with single-molecule localization microscopy (SMLM) under cryogenic conditions. Typically, these oligomers are assumed to consist of protomers arranged as equilateral two-dimensional polygons and every protomer is labeled with a dye molecule for visualization. Unlike previous work, we consider scenarios in which the sample plane has an unknown orientation relative to the focal plane. Our contribution is a high-precision plane-fitting algorithm to determine the sample plane, combined with geometrical transformations and two circle-fitting algorithms to identify the oligomeric structures. Our simulations on synthetic data demonstrate that the proposed workflow achieves high accuracy in estimating both the unknown tilted plane and the oligomer size.

Glucose-6-phosphate (G6P) is a key intermediate in multiple energetic and anabolic pathways, and quantifying its dynamics is essential for understanding cellular physiology. We have previously developed a syndicate of intensity-based, genetically encoded sensors based on the insertion of circularly permuted GFP into a Venus-flytrap-like analyte-binding protein. Here we use the same approach to develop an intensity-based G6P Sensing Fluorescent Reporter (iG6PSnFR). We present two variants: a G6P-activated sensor that increases fluorescence and a G6P-inactivated sensor that decreases fluorescence. We validate performance across progressively more complex preparations, including purified protein in vitro, immortalized and primary neuronal cultures, isolated pancreatic islets, in an intravital liver model, and finally in vivo in neurons. In each context, iG6PSnFR reports G6P changes consistent with expected responses to physiological perturbations.