Day 1: Fundamentals in Calcium Imaging
Day 1 of the ACCELERATE workshop began with general roundtable introductions and research interests from all of the attendees. While all of the scientists that participated in the event perform systems-based neuroscience, specific scientific interests included sleep and circadian rhythms, addictive behaviors (drugs and alcohol), learning and memory, appetitive taste behaviors, hypothalamic activity, as well as metabolism and obesity.
The goal of the 1st day session was to provide our scientists with solid fundamentals and steps to take in order to achieve a successful nVista user experience. Vania Cao, Ph.D., Inscopix’s Scientific Content and Training Manager and Camille de Solages, Ph.D., Inscopix’s Application Scientist each gave detailed presentations on the ideal nVista experimental workflow and a review of best practices in optimizing neuronal labeling using calcium (Ca2+) indicators. A number of critical steps are essential for imaging effective cell dynamics with the nVista system including stereotaxic precision, optimizing viral dilution and transfection, and careful planning of your experimental timeline. Vania also discussed various pros and cons to consider when choosing the appropriate lens for optical access along with useful surgical tips and guidelines to enable visualization of neuronal ensembles in different brain regions of interest.
The workshop then segued into a detailed review of the nVista software and set-up guidelines for performing imaging experiments led by Lara Cardy, Inscopix’s Customer Support Manager. The scientists were trained on how best to navigate multiple windows in the software, data collection, and optimal data management and file saving in order to achieve the best images for subsequent analysis. The 1st day workshop concluded with an overview of Inscopix’s Mosaic software, along with an example set of data to analyze, and was presented by our Application Engineer, Michele Jenkins. Michele provided everyone with a review on the ideal Mosaic workflow to streamline data analysis from image processing, motion correction, signal normalization, cell identification, and the use of PCA/ICA algorithms to enable accurate cell activity detection.
Day 2: Hands-on Calcium Imaging in Freely Behaving Subjects
Day 2 of the ACCELERATE workshop commenced with a trip to the Inscopix research laboratory. Srishti Gulati, Inscopix’s Lead Research Specialist, showed all of the attendees how to optimize their stereotaxic neurosurgical techniques to enhance live cell viewing of hundreds of neurons in the hippocampus through a lens probe. Many of the attending scientists asked questions and requested advice for optimizing their techniques related to their particular research interests. Chris Tsang, Ph.D., Inscopix’s Field Application Scientist then showed everyone how to optimally attach the nVista baseplate to ensure secure positioning of the miniature microscope. Some critical features of this procedure were reviewed in detail with all of the attendees such as ensuring that the microscope and baseplate are parallel to the face of the lens probe, along with proper positioning of the microscope’s focusing turret and gap distance between the objective and the lens.
The last live demonstration of the day showed the attendees how hundreds of neurons in the somatosensory cortex of mouse actively fired while the animal freely moved around in its home cage. Our Inscopix scientists also provided examples of how to optimize ongoing image and data collection using the nVista software.
The workshop concluded with a fun evening of food and drinks at Coconuts in Palo Alto and an opportunity for everyone involved to share their thoughts and experiences with the Inscopix platform. We thoroughly enjoyed having our valued Inscopix customers join us for our in-house workshop and we look forward to assisting them to achieve continued success!