Nanosecond Time-Resolved Fluorescence Assays

Abstract

Fluorescence-based assays are widely used in clinical diagnosis, drug development, environmental pollution detection, and chemical biology research due to their high sensitivity and rapid read-out times. Undesirable background fluorescence from complex matrices such as biological samples can be easily suppressed by time-resolved fluorescence (TRF) detection.Therein, long-lived probes are used and a delay time, after which all short-live background fluorescence has decayed, is applied to selectively detect the long-lived emission. In addition, to the established lanthanide-based probes with lifetimes in the millisecond time range, probes with much shorter lifetimes (>10 ns) can be used as well, for which the name Nano-TRF assays has been coined. This book chapter reviews the current state-of-the-art of Nano-TRF assays including probes based on Ru(II) complexes, pyrenes, and fluorazophores, as well as their applications.