ROBOTIC PLATFORM FOR THE DELIVERY OF GENE PRODUCTS INTO SINGLE CELLS IN ORGANOTYPIC SLICES OF THE DEVELOPING MOUSE BRAIN

摘要

Microinjection of genetic components and dye into organotypic slices provides excellent single cell resolution for unraveling biological complexities, but is extremely difficult and time consuming to perform manually resulting in low yield and low use in the developmental biology field. We developed a computer vision guided platform to inject specimen with mRNA, and/or dye and investigated the efficiency of the process using organotypic slices of the mouse developing neocortex. We demonstrate that the system significantly increases yield of injection relative to manual use by an order of magnitude, allows for cell tracking over 0, 24, and 48 hours post injection in culture, and enables mRNA translation of injected product. The autoinjector platform thus can open the door to new types of experiments including investigating effects of mRNA concentration, and composition on cell fate, and tracking these effects on cell reprogramming and lineage.