Multidimensional Robotic Flow Platform

We have developed a flexible, programmable, robotic flow platform that allows hands-free processing of biological and chemical samples. This approach not only allows high-throughput sample processing, but also accuracy, leading to fewer errors and more reproducible findings. The automated platform is currently being used to run simultaneously on the same samples (from healthy subjects or patients with autoimmune diseases) different projects for different investigators and postdocs interested in interrogating the immune system using multiparametric flow cytometry and gene expression. Polychromatic flow cytometric assays, using six to 18 different fluorochromes is a powerful tool to interrogate the cells of the immune system (fig A), ex vivo or after different conditions of culture on effects such as surface and intracellular markers, cytokine profile and differentiation, gene expression. Our aim is to apply this state of the art multidimensional robotic flow platform in a more robust and precise way to be used in fundamental and clinical research.

We have used validated methods on the Biomek FX and NX to thaw and immunophenotype (under sterile enclosure) batches of 24 or 48 coded West Nile and Type 1 diabetes samples before and after treatment. Samples were washed and counted using an automated counter (Guava) and the cell suspension adjusted to the appropriate concentration, processed and stained or stimulated and stained in a 96 well format using an automated platform. Data was acquired using 2 LSR Fortessa flow cytometers equipped with high throughput 96 well samplers, cell populations were also sorted using Aria. We use CST beads and specific application settings to minimize the variations that could be introduced by the flow cytometer laser performance on different days. We use automated compensation set on compbeads to standardize the flow data between different batches. To track the reproducibility of the assays and to monitor the acceptability of the %CV between different runs, one cryovial from a bank of frozen PBMC vials generated from single leukopack is processed and run simultaneously on the platform.

Current and future projects include: immune system phenogenetic profiling, ACE autoimmune diseases project, influenza vaccine project, different MS projects in the lab, collaborative projects with companies making new drugs to treat MS.