Diversity-Oriented Fluorescence Library Approach (DOFLA) for Discovery of Cell-Permeable Probes for Applications in Live Cell Imaging

Abstract

Fluorescent small-molecule probes can enter living cells and interact with molecules of interest to monitor specific organelle functions. The ideal imaging probes should be able to penetrate cellular membranes and should exhibit minimal background fluorescence signal in order to allow monitoring specific cellular functions without disrupting cell physiology or viability. In this chapter, we will introduce a powerful strategy, namely the diversity-oriented fluorescence library approach (DOFLA), which involves synthesizing a large number of fluorescent molecules using combinatorial chemistry. We present the concepts of DOFLA, DOFL construction techniques, and focus on the applications of cell-based DOFLA screening in probe discovery and subcellular behaviors analysis of small-molecule probes in living cells. This chapter highlights the advantages of DOFLA and provides a series of case studies of discovery of cell-permeable specific probes.