Single Quantum Dot Imaging in Living Cells

Direct visualization of biological processes at single-molecule level provides a detailed perspective which conventional bulk measurements are hard to achieve. Among various classes of fluorescent tags used in single-molecule tracking, quantum dots are particularly useful due to their unique photophysical properties. In this chapter, we describe the principles, methodologies, and experimental protocols for qdot-based single-molecule imaging. The first half provides an overview of fluorescent microscopy and advances in single-molecule tracking using quantum dots. The remainder of this chapter describes methods to carry out qdot-based single-molecule experiments. Detailed protocols including qdot labeling, microscopy setup, and single-molecule analysis using appropriate computational programs are given.