CalQuo: Automated, Simultaneous Single-Cell And Population-Level Quantification Of Intracellular Ca 2+ Responses
Detecting intracellular calcium signaling with fluorescent calcium indicator dyes is often coupled with microscopy techniques to follow the activation state of non-excitable cells, including lymphocytes. However, the analysis of global intracellular calcium responses both at the single-cell level and in large ensembles simultaneously has yet to be automated. We developed a new software package, CalQuo (Calcium Quantification), which allows the automated analysis and simultaneous monitoring of global fluorescent calcium reporter-based signaling responses in up to 1000 single cells per experiment, at temporal resolutions of sub-seconds to seconds. CalQuo quantifies the number and fraction of responding cells, the temporal dependence of calcium signaling and provides global and individual calcium-reporter fluorescence intensity profiles.
CalQuo is a MATLAB based software package developed at the University of Oxford. CalQuo is copyright protected and licensed with Isis Innovation (www.isis-innovation.com/). CalQuo allows the automated analysis and simultaneous monitoring of fluorescent calcium reporter based signaling responses in up to 1000 single cells per experiment. As output, CalQuo delivers the number and fraction of responding cells, their individual and characteristic calcium reporter fluorescence intensity profiles, and the highresolution temporal dependence of calcium signaling. CalQuo is well suited to studying other cell signaling processes, such as the release of proteins orother molecules, if these molecules can be detected using labels.
CalQuo2: Automated Fourier-Space, Population-Level Quantification Of Global Intracellular Calcium Responses
To expand upon the capabilities of CalQuo to recognize and quantify calcium responses in individual cells within a sample up to 1000 cells, we have developed an updated version, CalQuo2. Using the Fourier transform to convert calcium responses into the frequency space, CalQuo2 now requires only one (instead of five) user-defined parameter in determining a triggering from a non-triggering cell. Additionally, CalQuo2 can determine the fraction of cells that have a single peaked compared to the fraction of cells that have multi-peak calcium fluxes.
CalQuo2 is a MATLAB based software package developed at the University of Oxford. CalQuo is copyright protected and licensed with Isis Innovation. The new version employs Fourier-space analysis algorithm and is currently under review in a peer-reviewed journal.
Fritzsche M, Fernades RA, Colin-York H, Santos AM, Lee S, Lagerholm BC, Davis SJ, and Eggeling C, CalQuo: automated, simultaneous single-cell and population-level quantification of intracellular Ca 2+ responses., Scientific Reports, 2015.
Please contact for further information: marco.fritzsche (at) rdm.ox.ac.uk or ricardfe (at) stanford.edu.
Please contact for further information: marco.fritzsche (at) rdm.ox.ac.uk.