Abstract: In recent years novel single-cell sequencing methods have allowed an in-depth analysis of the diversity of cell types and states in a wide range of organisms. Due to the continuous optimization of experimental and computational methods by many research groups, it is now possible to sequence the transcriptomes of thousands to millions of individual cells. Albeit an exciting development, transcription only covers the first step in the central dogma. The second step, the process of translation, is currently much harder to explore in single cells. Despite recent progress in detecting proteins by mass spectrometry with single-cell resolution, it remains a major challenge to measure translation in individual cells. Building upon existing ribosome profiling protocols our laboratory recently increased the sensitivity of these assays allowing ribosome profiling in single cells. I will present our ongoing efforts to determine translation efficiency in single cells and to correlate translation efficiency to tRNA levels and tRNA modifications. Additionally, I will explore how these approaches could provide insights into ribosome assembly and ribosome structure at the level of an individual cell.