Confocal Microscopy of Diverse Protocells

Modern life is composed of compartmentalization, genetics, metabolism, and self- replication. The precursors to all of this are thought to be protocells, simpler cells that have a lipid membrane with RNA on the inside. This report focuses on the lipid membrane aspect of protocells. Protocells can be made spontaneously if the right environmental conditions are met (eg. pH and concentration of chemicals). We made these protocells in the lab as a model system for the origin of cellular life. Different environmental conditions can make protocells with different membrane characteristics (eg. number of layers, shape, and size). We analyzed these protocells by staining the membrane and inside of the protocells with dyes and imaging them on a confocal microscope. The confocal microscope gives high-resolution in speed, pixel resolution, and allows for videos. To help image accurately, we utilized biotin- avidin conjugations to anchor protocells to the glass surface. Through our confocal microscopy experiments, we observed several complex protocell morphologies and shape transformations, these give new insights into how diverse protocell shapes have different functions.