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CytoTRACK SPAchip® pH Dual detection kit
CytoTRACK SPAchip® pH Dual detection kit
Ref. D-001-PHGR
CytoTRACK SPAchip® pH Dual Detection Kit is the combination of the two technologies addressed to measure intracellular pH: CytoCHECK SPAchip® pH red single-detection andCytoCHECK SPAchip® pH green single-detection.
The product combines these two pH detection technologies so that each of them is internalized in one cell line for co-culturing applications. While tracking the two different cell subtypes and their interaction, measurements of intracellular pH levels by changes in fluorescence intensity are feasible at real time.
The advantages of this product are the combination of our two CytoCHECK pH detection kits, where the selection of the SPAchip® to the corresponding cell line will depend on the features of each. Red emission SPAchip® is recommended for applications which need to overcome high green autofluorescence signals and increase SNR. Green emission SPAchip® advantages are its easiness due to the possibility of using common GFP fluorescence filters and its easy linear response.
Kit box contains:
2 ASSAY SPAchip® eppendorf tube (Quantity: 2.5 x10^6 ASSAY SPAchip®)
1 Control SPAchip® eppendorf tube (Quantity: 5 x10^5 CONTROL SPAchip®)
1 buffer (Quantity: 8mL)
Quick start guide
Brochure
User protocol
Key features
- Non-toxic for living single cells.
- Measures intracellular pH levels by changes in fluorescence intensity.
- Allows long-term monitoring of intracellular pH changes
- 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.
- Cell type flexibility, no lower limits.
- Ready-to-use, robust workflow
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 | Green Fluorescence |
| Fluorescence | λex: 546 nm; λem: 610 and 707 nm | λex:488 nm; λem: 520 nm |
| For research use only | Not for use in diagnostic procedures | |
| Incubation time | Overnight | |
| Measuring range | pH 4.5 – 9.0 | pH 4.5 – 7.5 |
| 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 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.
Figure 1: : Red and green CytoTRACK® pH Detection KIT at different pH conditions using commercial calibrators inside cells.
A) Graph showingratiometric normalized fluorescence intensity values of intracellular CytoCHECK SPAchip® pH RED Single Detection at different pH conditionsusing intracellular calibrators in HeLa cells. Ratiometric values were obtained by dividing λem2=707/ λem1=610 nm emission signals in HCSOperetta equipment with the excitation in the range λex=546/15 nm.
B) Graph showing normalized relative fluorescence intensity values ofintracellular CytoCHECK SPAchip® pH GREEN Single Detection at different pH conditions using intracellular calibrators in CAL51 cells.
Like an eye
in the cell!
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