The Zebrafish testing site includes experts in genetic engineering and live optimal imaging (confocal, multi-photon, video-wide field) in zebrafish. The team is specialized in structural and functional multi-plane whole zebrafish brain imaging with single neuron resolution, as well as synaptic imaging in individual neurons in zebrafish brain. Eggs are injected with the gene(s) of interest upon fertilization with plasmids encoding various sensors, such as those developed by our colleagues from the Protein Engineering Core. Tests are conducted to verify their expression, functionality, and properties; providing a rapid and critical feedback on the suitability of the newly developed tools for in vivo applications in this model.

The Tadpole (Xenopus) testing site has been studying Xenopus neurodevelopment for two decades.  The testing model starts with any plasmid that bears the sequence of a novel fluorescent protein (FP) or reporter.  The gene is subcloned into an mRNA expression vector, mRNA synthesized in vitro, and microinjected into in vitro fertilized Xenopus embryos at the one- or two-cell stage of development.  This typically results in rapid whole-animal expression of the gene of interest.  Using in vivo 2-photon microscopy, the function of the gene product can be assessed in any organ of the rapidly developing embryo – typically the central nervous system – and its activity quantitatively compared to other benchmark FPs such as GCaMP6s.  The entire pipeline from DNA sequence to in vivo CNS characterization takes approximately two weeks.

The Invertebrate Testing Site is largely based on the fruitfly (Drosophila) model. Drosophila larvae have a rich repertoire of behaviors, responding to specific stimuli and using various strategies to escape from negative stimuli.  Escape behaviors are critical for survival and require optimal performance.  To achieve such performance, the larval brain must produce specific spatiotemporal patterns of output appropriate to the corresponding patterns of input.  Such decisions require multisensory integration—a process that is difficult to study in a large nervous system because the convergence of neural signals must be tracked at many sites. The neuron-behavior relationships analysis combines 1) genetic tools to manipulate individual neurons; 2) a high-throughput behavioral tracking system that allows temporally controlled stimulation of many freely moving larvae at once; 3) TEM neuron reconstruction; and 4) unsupervised structure learning methods to categorize behaviors in an unbiased fashion.

The IPSC-Derived Neuronal Cultures testing site relies on a team of highly qualified personnel, and comprises a park of multimodal microscopes allowing the high-content label-free testing and optimization of the optogenetic tools that are designed, produced, and packaged by colleagues of the protein engineering and viral vector cores of the COVF . Those tests are conducted on cortical neural networks derived from a biobank of highly phenotyped human induced pluripotent cell lines (hiPSCs).

The IPSC-derived Organoids testing node is part of the Early Drug Discovery Unit (EDDU) at The Neuro. The EDDU specializes in generating different types of brain cells and 3D brain organoids for fundamental and translational discovery, with workflows and the infrastructure available for automation, small molecule screening and single cell phenotyping by flow cytometry. Together with Canadian and international academic and industrial partners, we work with high quality patient-derived, gene-edited, and isogenic control iPSC-derived cells to build new disease-relevant tools and assays for discovery purposes. With a rapidly growing base of both active academic and industry users, the EDDU has grown steadily over the past six years, slowly becoming a hub for training HQP from labs across Canada on everything from iPSCs to discovery assays.

The Live Human Dorsal Root Ganglion and Spinal Cord Tissue is located at McGill. It is equipped for testing viral transduction and optogenetics tools in live human dorsal root ganglion and spinal cord tissue harvested thanks to a partnership between the Alan-Edwards Centre for Research on Pain at McGill and Transplant Quebec through a consortium of organ donors across several hospitals in Montreal.

Located at UBC Djavad Mowafaghian Centre for Brain Health, the home-cage Home-Cage Mesoscopic Functional Imaging of Mouse Cortex site provides an in vivo testing platform specializing in fiber photometry, which allows probes to be assessed in specific brain regions/nuclei during behavioral engagement, in addition to expertise in mesoscale cortical widefield imaging. We combine these imaging techniques with automated home cage motor assessment to provide high throughput testing of new reagents and animal phenotyping in models of human neurological disease.  New infrastructure in 2022 will expand our testing node to include new modalities: 3-photon imaging and mesoscale cellular imaging with ultra-wide field of view 2-photon imaging.  A strong training component exists within the NeuroImaging and NeuroComputation Center, where a weekly technology forum — Databinge, meets to discuss and implement data analysis approaches and advanced training.  We are currently working to extend Databinge to a multi-institution platform through interactions with the Canadian Brain Research Strategy, other national groups and our international partners in the International Network for BioInspired Computing (USA, France, Canada).  Recent success in CFI competitions has led to the development of a multi-scale imaging platform termed iMAP that provides testing across multiple scales of resolution. Recent development of open science principles ensure sharing of collected data and increase transparency in research.

Located at the Hotchkiss Brain Research Center (University of Calgary), the Photometry/Optogenetics of Neuroendocrine Signalling facility is composed of an extended team of experts in rodent stereotaxic brain surgery, in vivo validation, fibre photometry using calcium or other biosensors, optogenetic stimulation or inhibition, in vivo cellular calcium imaging and a variety of behavioural procedures and computational methods. The Borgland lab is a member of the optogenetic core facility and has expertise in validating viral vectors using immunohistochemistry, brain slice electrophysiology, and in vivo behavioural models with a focus on reward learning. The Borgland lab is a COVF testing site and has used several viral vector tools disseminated from the Laval core facility to test in rodent in vitro brain slice preparation to test synaptic connectivity and in in vivo behavioural models of reward learning. Results from novel vectors are shared with the COVF team.

Located in uOttawa’s Faculty of Medicine (FOM), the Synaptic Release and Plasticity testing site is part of the Cell Biology and Image Acquisition core (CBIA); (https://med.uottawa.ca/core-facilities/facilities/cbia) it oversees the use and maintenance of several optical imaging systems (eg., Confocal and multiphoton microscopes; STED system; Spinning disk; imaging software; analysis workstations and others). The CBIA facility’s operating budget (>$300K per annum) is based on a partial cost-recovery model, with direct institutional contributions from uOttawa and the FOM that allows competitive user fees to the community. The CBIA facility has a broad mandate, including covering service contracts for major equipment, overseeing a preventative maintenance program, providing optical and imaging training to HQPs and providing funds for the procurement of new instruments or replacement parts. The CBIA is operating under a Planning and Priorities Committee and is staffed by a Senior Imaging specialist and an imaging technician that carry out the day-to-day activity. The Béïque lab will act as the formal contact between this core, uOttawa’s neuroscience community and members of the COVF. His lab has developed expertise in employing cellular electrophysiological techniques in combination with several optical approaches (optogenetics and multiphoton imaging) to study neuronal dynamics over different time scales. The uOttawa testing site will provide experimental testing of viral vectors and optical sensors in rodents both in vivo (cellular imaging in learning tasks by 2P or miniendoscope imaging), and in vitro (electrophysiological calibration of 1 or 2P signals). The performance of the newly developed sensors will be further examined by concurrent advanced analytical and network simulations tools by the computational neuroscientists at uOttawa’s Center for Neural Dynamics.

Located at the CERVO Brain Research Centre (Université Laval), the spinal optogenetics site is composed of an extended team of experts in optogenetics, imaging and electrophysiological approaches in both anaesthetized and freely moving animal. Our goal is to offer to every researcher requiring our help a testing node to certify the efficiency of their hardware or molecular tools for optogenetic experiments. To do so, our core is specialized in the acquisition of calcium signals, optogenetic mediated neuronal manipulation with innovative technologies such as in vivo micro-endoscopy, photometry combined or not with electrophysiological recording. Our expertise allows us to acquire and analyze signal from a wide range of brain regions down to the cellular resolution in rodents. By working in close collaboration with the viral vectors platform we can validate custom-tailored tools before implementation in larger scale experiments thus further nurturing the feedback loop between production site and customers, ensuring the constant improvement of the tools developed.

The facility allows access to cutting-edge in vivo infrastructure and technical know-how for McGill, Canadian, and international researchers engaged in neurophysiology, neuroimaging, behavioural analysis, and disease modeling to effect meaningful translational neuroscience research. A main goal of the platform is to close the ‘translational gap’ between rodent and human species through the utilization of the common marmoset (Callithrix jacchus) model. The complex brain anatomy, cognition, and behaviour of marmosets and their amenability for genetic engineering make them a powerful model for understanding brain activity underlying behaviour and for confronting brain disorders/diseases. The MGH platform is making accessible cutting-edge viral delivery approaches and in vivo cellular imaging using miniscope hardware in marmosets. Coupled to the marmoset imaging aspect of the platform will be the establishment of neurophenotyping capacity involving behavioural testing, movement tracking, and pharmacokinetic analysis for drug studies. Furthermore, the platform is also developing services for robotic brain injections, magnetic resonance imaging, and pharmacokinetic analysis in marmosets. Thus, the MGH testing site has important capacity for investigating neural activity in the primate brain using optogenetic tools developed by COVF and offers unique services that do not exist elsewhere in Canada.