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The Gordon Conference on Single Molecule Approaches to Biology covers the rapidly evolving field of development and application of single molecule techniques to biological problems. Single molecule techniques provide previously unobtainable data on fundamental biochemical processes that are essential for all forms of life, including human and human pathogens. Recently, tremendous technical developments have made it possible to detect, identify, track, and manipulate single biomolecules in an ambient environment or even in a live cell. Single molecule approaches have changed the way many biological problems are addressed. New knowledge derived from these approaches continues to emerge. The ability of single molecule approaches to avoid ensemble averaging and to capture transient intermediates and heterogeneous behavior renders them particularly powerful in elucidating mechanisms of molecular machineries: what they do, how they work individually, how they work together, and finally, how they work inside live cells. The burgeoning use of single molecule approaches to elucidate biological problems is a highly multidisciplinary pursuit. This conference seeks to bring together top experts in molecular and cell biology with innovators in the measurement and manipulation of single molecules. The intention is to better promote the use of the single-molecule approach to generate new biological knowledge and to better orient the development and implementation of single molecule techniques for compelling biological problems.