Cell signaling cascades resulting from interactions between G-protein coupled receptors (GPCRs) and their ligands control a wide variety of intracellular processes, making them major targets of drug discovery efforts, particularly for cancer treatment. The cell signaling cascade takes the form of a complex, multifaceted series of second-messenger systems that act in concert. One important second messenger in GPCR-mediated cell signaling is calcium (Ca2+), which plays a critical role in cell response to intracellular and environmental cues. The ability to detect rapid changes in intracellular levels of calcium with high temporal resolution is key in characterizing GPCR activation.
BioTek recently launched the Lionheart™ FX Automated Microscope – a compact, inclusive microscopy system – ideal for the calcium imaging workflow. In this article, SelectScience® speaks with Dr. Peter Banks, Scientific Director of BioTek Instruments, Inc. to learn how the company’s latest cell imaging technology can advance calcium flux analysis, by imaging individual cells with sub-second temporal resolution.
We are primarily interested in calcium mobilization as it pertains to G protein-coupled receptor (GPCR) activation. This gene family has been and remains one of the most actively pursued for drug discovery. Many different types of instruments have been used to detect calcium mobilization, including microplate readers measuring fluorescence intensity with dyes like Fluo-4. Imaging which enables the visualization of each cell in the field of view, used in conjunction with sophisticated image analysis routines, allows for the segregation of cells that respond fully to GPCR activation, providing greater assay performance than that achieved using an average response of all cells.
Tell us about some of the features of the Lionheart that have helped to advance calcium imaging?
What feedback have you had from scientists about using the Lionheart for calcium mobilization?
What do you see for the future of GPCR signaling pathway analysis and the technologies involved?
GPCRs use multiple pathways for cellular signaling involving a number of second messengers. Being able to simultaneously measure transient responses in live cells from multiple second messengers will provide a much greater understanding of GPCR signaling and this is where I think the future of GPCR analysis is heading. BioTek is currently collaborating with a reagent company called Montana Molecular that provides GPCR biosensors for many GPCR second messengers, including calcium, cyclic-AMP, diacyl glycerol and PIP2. These biosensors are provided with different spectral characteristics that allow for the multiplexed analysis of GPCR second messengers.
How do you hope BioTek technology will advance drug development/treatments for disease?
BioTek’s Automated Cell Imaging product line brings highly flexible, cost-effective cellular imaging to just about any laboratory, whether it’s those in big budgeted Pharma, the more cost-conscious academic lab or spin-off start-up. We hope to enable a wide range of customers interested in better understanding disease onset, progression, diagnosis and treatment.
Learn more about the cell imaging technology available from BioTek and how it can be used to characterize calcium flux.