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CytoCHECK SPAchip® pH red single detection kit

CytoCHECK SPAchip® pH red single detection kit

Ref. S-001-PHR

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 in the cytosol of cultured cells to monitor intracellular parameters for extended periods of time.t

CytoCHECK SPAchip® pH RED Single-Detection Kit allows measurement of intracellular pH 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. Its signal in red makes it feasible for applications with high green autofluorescence such as organoids cultures. Moreover, due to its ratiometric behaviour signal to noise ratio is increased leading to cleaner curves.

Kit box contains: 

1 ASSAY SPAchip® eppendorf tube (Quantity: 2.5 x10^6 ASSAY SPAchip®)
1 Control SPAchip® eppendorf tube (Quantity: 2.5 x10^5 CONTROL SPAchip®)
1 buffer (Quantity: 5mL)
Quick start guide

Brochure

PRODUCT Sheet

User protocol

Key features

Measures intracellular pH 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 pH changes.

Technical specifications

Amount	~2.5 millions of SPAchips
Applications	Cell Viability, proliferation, cell image acquisition
Assay Time	30 minutes
Assay type	Living single-cell based
Detection method	Red Fluorescence
Fluorescence	λex: 546 nm; λem: 610 and 707 nm.
For research use only	Not for use in diagnostic procedures
Incubation time	Overnight
Measuring range	pH 4.5 – 9.0
Platform	Fluorescence microscopy, HCS/HCA platforms and flow cytometry
Sample type	Adherent cells, suspension cells
Shelf Life	6 months upon arrival
Shipping	Environmental conditions
Solubility	Soluble in assay buffer (aqueous)
State	Solid-phase film with embedded SPAchips
Storage Conditions	Refrigerate upon arrival (2ºC to 8ºC) and protect from light

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 546 nm and detecting fluorescence emission at 610 and 707 nm. Normalization of fluorescence intensity values collected at both emission wavelengths provide a ratiometric behavior that allows a more precise quantification of intracellular pH. 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.

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.