Calcium Flux

G protein coupled receptors (GPCRs) are among the most heavily investigated drug targets in the pharmaceutical industry, and Calcium flux is a key functional assay for monitoring GPCR activation.

High Content Analysis is a powerful tool for image-based quantification of drug induced GPCR activation. It provides cell by cell analysis leading to reliable data independent of the transfection rate for transiently transfected cells. It also generates additional readout parameters for calcium response, such as the number of responders in a population. Compared to systems that average the response across the entire population within a well, such approaches can significantly decrease the number of false positives identified.

Schematic showing labeling of intracellular free calcium upon glutamate-induced activation of G-protein coupled receptor stimulation by glutamate. Calcium ions are detected using the calcium sensitive dye, Fluo-4. The graph shows the time course of Fluo-4 mean fluorescence intensity for the whole cell population (purple), for the active (yellow) and inactive (gray) cell subpopulations .

Our Calcium Flux solutions

Drawing on many years of experience and in-depth knowledge, PerkinElmer offers solutions for every throughput, a starting point for typical transcription factors assays and unlimited possibilities for customization:

Opera High Content Screening System

• On-board non-contact dispenser for reagent addition and fast mixing, 0.5 - 10 µl dispensing for assays in 96-well, 384-well and 1536-well format
• Measurement of multiple wells in parallel (dispenser moves independently from scanning stage for nested kinetic scenarios)
• Variable time points for kinetics – short intervals at the beginning of a reaction and longer intervals at later stage
• Full environmental control (temperature, CO2 and humidity)

Acapella Image Analysis Software

• Image analysis sequences for calcium flux, single cell readout 
• Population based readout allows separation of responders and non-responders, for example

cell::explorer Automated High Content Screening System

• Full environmental control and workstation with hand-over of plates from imager to incubator for short-term and long-term live cell assays
• Scalable and fully automated workstation including compound handling, reagent addition, incubation, cell fixation and staining
• Instruments with readout other than imaging can be included, such as whole well fluorescence / luminescence (e.g. EnVision, EnSpire)

You may also be interested in: Live Cell Imaging.