CytoCHECK SPAchip® calcium single detection kit
CytoCHEK SPAchip® assay kits are novel fluorescence cell- based assays developed by A4cell that brings together the fields of nanotechnology and cell biology. SPAchips are composed of fluorescently labeled silicon microparticles which can be internalized inside single cells for lengthy periods, providing highly accurate data through long-term cell monitoring.
CytoCHECK SPAchip ® Calcium Single-Detection Kit allows measurement of intracellular calcium levels by changes in fluorescence intensity, which allows a more comprehensive study of the living single-cell physiology and maximizes the performance of most of imaging analyzers.
Measures intracellular calcium levels by changes in fluorescence intensity.
Composed of fluorescently labeled silicon microparticles that can be internalized in the cytosol of cultured cells.
Provides a more comprehensive study of single-cell physiology and metabolism.
Maximizes the performance of most imaging analyzers.
Non-invasive and allows long-term monitoring of intracellular calciumchanges.
Imaging and fluorescence signal analysis
SPAchip® kits are designed to dynamically shift fluorescence intensity values in response to intracellular changes in analyte concentration. Fluorescence intensity is measured by exciting the live cell sample at 488 nm and detecting fluorescence emission at 520 nm. Relative intensity units depend on the instrumental setup (excitation power, detector efficiency, etc). To obtain quantitative data, a calibration curve needs to be analyzed in the same run. Data acquisition can be adapted to fluorescence microscopy and flow cytometry. Additional detection channels, such as brightfield, phase contrast, FSC or SSC, are suggested to better identify individual SPAchips.
a) Emission spectra of Calcium--sensing probes when excited at the indicated wavelength. b) Brightfield image of SPAchips® once in solution (Image from Torras et al. 2015 DOI: 10.1002/adma.201504164), Scale bar = 10 µm. c) Calibration Curve of Fluorescence Intensity values from CytoCHECK SPAchip® Calcium Single Detection kit at different concentrations.
SPAchip® fluorescence signal quantification can be adapted to any analysis image software. Object segmentation should be performed to identify SPAchips inside cellular region to every sample. Fluorescence intensity units can be exported as a spreadsheet to plot graphical summaries of the results. Quantitative data need to be interpolated from the calibration curve values to each experiment. Normalization of results to experimental controls offer a semi-quantitative approach.
Image Analysis Workflow. From sample data acquisition to object identification, quantification and result presentation.
CytoCHECK SPAchip ® calcium single detection kit box contains:
• 1 ASSAY SPAchip® eppendorf tube (Quantity: 2.5*10^6 ASSAY SPAchip®)
• 1 Control SPAchip® eppendorf tube (Quantity: 2.5*10^5 CONTROL SPAchip®)
• 1 buffer (Quantity: 5mL)
• Quick start guide