The capability to distinguish the unique behavior of individual cells within tissues has enabled progress in many fields of biology. This new single cell perspective has the potential to transform our understanding of many critical biological processes and diseases, from development and regeneration to cancer. While the field of single cell biology has shown enormous growth in technical capabilities over the last decade, many conceptual questions remain, such as how to best apply and develop these technologies to answer key biological questions. Key areas of biological interest include: (1) Developmental processes-- how do individual cells differentiate to generate cell types and structures that comprise adult organisms? (2) Tissue ecosystems-- how cells interact with and respond to their neighbors within organized tissues to support tissue function and homeostasis? (3) Disease--how does the misbehavior of single cells impact disease development and manifestation in cancer and other ailments? (4) Technology Development—what kinds of new data collection and analysis tools are needed to fill current biological knowledge gaps? The mission of this meeting is to bring together researchers who develop single cell biological tools, with those from neighboring fields who are utilizing these technologies, to address these questions and derive new ones in a collaborative manner. Cross-talk between these disciplines will identify new opportunities for both technology development, and applications to answer novel biological questions, that will accelerate advances across fields. Discussions and collaborations seeded at this meeting will drive the field in innovative and fruitful directions long into the future, with far more powerful impacts than could be achieved in isolation.