CDg4, a novel green fluorescent mESC probe was discovered through a high-content image based screening of 160 members of the chalcone library. Interestingly, the molecular binding target of CDg4 was identified as the glycogen of the stem cell colony surface, rather than a conventional protein target from an intracellular source.

To investigate intracellular binders of myotube‐specific probes, a thiol‐reactive derivative, CDy2, was prepared. In live cells, the derivative selectively localizes in mitochondria and covalently labels its binding partners.

The first BODIPY library (BD) was synthesized, and a highly selective glucagon sensor, Glucagon Yellow (BD-105), was discovered by fluorescence image-based screening method. Glucagon Yellow exhibited selective staining only in AlphaTC1 cells. The selectivity of Glucagon Yellow toward glucagon was confirmed in vitro by comparison of its fluorescence intensity change against 19 biologically relevant analytes.

Heparin Blue was developed by a diversity-oriented fluorescence library approach (DOFLA) from a benzimidazolium library; the discovered compound showed remarkable properties and have the potential to be applied to monitoring heparin levels in clinical plasma samples for point-of-care detection.

A fluorescent dye library approach for the development of a bioanalyte sensor was sought. The screening of a rosamine dye library against diverse macromolecules led to the discovery of a highly sensitive human serum albumin binder, G13, with ∼36-fold fluorescence intensity change. G13 showed a highly selective response to HSA over other macromolecules including albumins from other species. The potential use of G13 for the detection of HSA in biofluids is described.

A group of styryl‐based neutral compounds has been developed as potential imaging agent candidates for the β‐amyloid plaques seen in Alzheimer’s disease (AD). The representative compound STB‐8 successfully penetrated the blood-brain barrier and specifically stained amyloid plaques of AD model transgenic mice ex vivo and in vivo.

Combinatorial approach on fluorescent rosamine library was developed using solid-phase chemistry. A highly selective library member (H22) toward reduced glutathione (GSH) was discovered, demonstrating the capability of in vivo GSH level sensing in live cells.

This protocol outlines a methodology for the preparation and characterization of three RNA-specific fluorescent probes (E36, E144 and F22) and their use in live cell imaging. It describes a detailed procedure for their chemical synthesis and purification; serial product characterization and quality control tests, including measurements of their fluorescence properties in solution, measurement of RNA specificity and analysis of cellular toxicity; and live cell staining and counterstaining with Hoechst or DAPI. Preparation and application of these RNA imaging probes take 1 week.

Highly selective fluorescence turn-on GTP sensor, GTP Green, was discovered by a diversity directed sensor approach, combined by solid-phase combinatorial synthesis of a benzimidazolium library and high-throughput screening.

A library of fluorescent styryl dyes (320 compounds) was prepared by solid‐phase chemistry. The dyes were screened for their detection of amyloid aggregates, which are associated with diseases such as Alzheimer’s, and two of the 320 compounds screened, 2C40 and 2E10, showed promise as brain‐imaging agents.