Biophysics at the single-molecule level provides the means to study complex, heterogeneous biological systems and to overcome many hurdles that are common to ensemble studies. Single-molecule biophysics has therefore continued to be one of the most exciting and powerful scientific disciplines in the life sciences.
Throughout this course, we will show how single-molecule methods uncover mechanistic insights that are often inaccessible by bulk techniques and explore their growing impact on central biophysical discoveries.
The course will begin with a lecture or two that outline the motivation and evolution of single-molecule experiments. This will be followed by a discussion and analysis of key papers in the field, which we will treat as case studies highlighting the use of single-molecule approaches to investigate fundamental biological processes.
The papers will be presented by the students, and each presentation will be followed by a general assessment of the contribution and importance of the work to the field.
In the discussion, we will emphasize two important aspects of the papers:
- Applications of single-molecule techniques to biological questions, including protein-DNA interactions, protein folding dynamics, enzymatic activity, the mechanisms underlying molecular motor action, and more. The significant advantage of single-molecule studies in deciphering biological dynamic processes will be highlighted.
- Single-molecule detection and manipulation techniques. We will analyze key experimental approaches, including fluorescence-based methods (e.g., single-molecule FRET, total internal reflection microscopy), force spectroscopy (optical and magnetic tweezers, AFM), fluorescence labeling strategies, and data analysis workflows. Emphasis will be placed on the principles, instrumentation, and practical challenges of each technique.