2. TRD#1: New Approaches to Specimen Preparation Abstract We are developing novel methods for specimen preparation that provide several advantages over traditional methods. These advantages include consumption of very small amounts of sample, and providing for more control over the quality of the prepared grids. The system uses a robotic device, called Spotiton, to dispense drops with volumes on the order of nanoliters to precise locations on an EM grid. The success of the device is coupled to a new “self-blotting” grid that we have developed to provide a method for spreading sample to a thin film without the use of externally applied filter paper. Over the next 5 years we will design, develop, test and validate a generalized Spotiton system capable of multiple tasks including making well controlled vitrified grids, managing time resolved and fast mixing experiments, and preparing ~100 independent negatively stained samples on a single grid. We anticipate that a version of this device will be manufactured by a commercial company so that it becomes generally available to the research community.