Automated Robotic Microscopy System Combining Longitudinal Single Cell Image Analysis with High Throughput/ High Content Screening. INVENTORS

SUMMARY Gladstone scientists developed automated robotic microscope technology that combines the sensitivity and resolution of longitudinal single cell analysis with the power and capacity of high throughput/ high content screening (HTS/HCS). HTS/HCS is a mainstay of the drug discovery process and our systems provide unique additional power over other commercially available arrangements. Most recently, our computer controlled automated robotic microscopy system has undergone a series of modifications to improve HTS/HCS analysis of biological samples (cells or tissues), while reducing user intervention relative to existing technologies. New configurations facilitate a precise return to, and re-imaging of, the same field (e.g. the same cell). The increased capabilities of the system enable sophisticated experimental designs to test hypotheses dealing with causality over time with greater precision and throughput than existing technologies. Our system incorporates computer software and hardware for control of the chosen microscope system (or other specific imaging device), a bulk sample storage subsystem and a robotic arm (or alternative transport device). The robotic arm can be programmed to select barcoded samples within the storage subsystem, then transport and place these samples in the sample holder on the imaging device. The robotic arm can transport samples of varied size using adjustable grippers. Fiduciary marks on the samples enable a precise automatic alignment within the sample holder by matching up with marks from previously positioned samples. Software programs orchestrate fully automated image acquisition and analysis; stitching together images of microscope fields taken from each single well of a micro-titer plate. These montages are temporally organized then used to assign each cell the equivalent of a “social security number”. When fluorescent reporter proteins are introduced into live cells to probe the activity of biological pathways of interest, large data sets can be automatically analyzed at great speed with the same rigorous statistical tools used for clinical studies. Highly quantitative cell biology with unprecedented sensitivity is automated using the robotic microscope, such that a study of 300,000 cells in a typical experiment takes only 15 minutes to analyze. The criteria used to identify features of interest in cells are explicitly defined and uniformly applied by the software, eliminating any user bias. This automated robotic microscope system has been applied with great effect in primary cells exhibiting time dependent stochastic cell changes relating to normal physiology and disease states in neurodegeneration.