High‐Throughput Confocal Microscopy for β‐Arrestin–Green Fluorescent Protein Translocation G Protein‐Coupled Receptor Assays Using the Evotec Opera

Translation of a Tumor Microenvironment Mimicking 3D Tumor Growth Co-culture Assay Platform to High-Content Screening

“…For automated fluorescent image acquisition, the Opera QEHS (PerkinElmer, Waltham, MA), [15][16][17] the MIAS-2 (Digilab, Holliston, MA), 18 or the IN Cell Analyzer 2000 (GE Healthcare Life Sciences, Little Chalfont, UK) 19 was used. The MIAS-2 and IN Cell 2000 can also acquire brightfield images.…”

“…The PerkinElmer Opera QEHS [15][16][17] is a microlensenhanced Nipkow spinning disk confocal system for fluorescence imaging. It is powered by four lasers and images of three colors can be acquired in parallel by the three cameras.…”

Scintillation proximity assay in lead discovery

“…Optimization of the Transfluor assay for HTS applications is growing in popularity but will require more time to demonstrate its full potential in a high-throughput setting [58,59] . The demands for functional assays to measure GPCRmediated second messenger systems (e.g., IP 3 , calcium, cAMP) are now achievable through a variety of fluorescencebased approaches of which many can be fully automated for robotic fluid dispensing and HTS.…”

The Roles of High Content Cellular Imaging in Lead Optimization

“…Recently, HCCI has been effectively utilized in conjunction with a genomics approach for target identification and validation of orphan GPCRs. In this study, bioinformatics and tissue dependent expression of orphan GPCR mRNA was used in conjunction with the G-protein pathway independent b-arrestin-GFP assay to identify, prioritize, screen and characterize orphan GPCRs related to obesity (9). The multiplex readout capability of HCCI has been used to measure the activity of the mitogen activated protein kinases (MAPK), p38, ERK, and JNK and related biology such as receptor internalization, transcription factor translocation, and cytotoxicity (12 -20).…”