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CytoCHECK SPAchip® OHrad ROS Single-Detection Kit
CytoCHECK SPAchip® OHrad ROS Single-Detection Kit
Ref. S-003-ROSG
CytoCHECK SPAchip® OHrad ROS Single-Detection Kit allows real-time monitoring of intracellular reactive oxygen species (ROS) through the specific detection of hydroxyl radicals (•OH) via changes in green fluorescence intensity within living cells. This product enables to measure •OH radicals generated either as primary ROS or as secondary species downstream of other ROS, facilitating a more comprehensive study of live cell physiology, proliferation, differentiation, cell death, and ferroptosis.
CytoCHECK SPAchip® OHrad ROS Single-Detection Kit is pH-insensitive: experimental readouts obtained from this product are not affected by pH changes of the media, avoiding measurement artifacts due to pH variations.
CytoCHECK SPAchip® OHrad ROS Kit is only active when in contact with intracellular esterases, obtaining a fluorescent signal with internalized SPAchips.
SPAchip® OHrad ROS technology enables continuous and accurate monitoring of intracellular ROS levels in living cells, by the specific detection of hydroxyl radicals (•OH), without any perturbation of cell physiology or metabolic state over long periods of time. Thus, CytoCHECK SPAchip® OHrad ROS Single-Detection Kit allows the tracking of important biological processes, helping to develop and improve strategies for anti-cancer therapy assays.
Kit box contains:
1 ASSAY SPAchip® eppendorf tube (Quantity: 2.5 x10^6 ASSAY SPAchip®)
1 Control SPAchip® eppendorf tube (Quantity: 2.5 x10^6 CONTROL SPAchip®/mL)
1 ASSAY buffer tube (Quantity: 5mL)
Quick start guide
DATA SHEET
APPLICATION NOTE
User protocol
Highlights
- Intracellular measurements of reactive oxygen species (ROS) levels (specifically hydroxyl radicals) by changes in fluorescence intensity.
- Designed to be active only when in contact with intracellular esterases.
- This product is pH-insensitive.
- Non-invasive for live cells allowing long-term monitoring of intracellular ROS changes.
- Composed of fluorescently labeled silicon microparticles that can be internalized in the cytosol
of cultured cells. - Ready-to-use workflow that provides an extensive study of cell physiology and metabolism.
Key features
- CytoCHECK SPAchip® kits are novel cell-based assays for living cells that bring together the fields of nanotechnology and cell biology.
- CytoCHECK SPAchip® detection kits are composed of fluorescently labeled silicon microparticles -SPAchips®- that can be internalized in cultured cells to monitor changes in specific intracellular analyte concentrations for long periods of time.
- This product enables a more comprehensive study of cell survival, proliferation, differentiation, cell death, apoptosis, and ferroptosis.
- CytoCHECK SPAchip® OHrad ROS Single-Detection Kit enables continuous, and accurate monitoring of intracellular ROS levels in living cells, by the specific detection of hydroxyl radicals (•OH).
- CytoCHECK SPAchip® OHrad ROS Single-Detection Kit is pH-insensitive: experimental readouts obtained from this product are not affected by pH changes of the media, avoiding measurement artifacts due to pH variations.
- CytoCHECK SPAchip® OHrad ROS Kit is only active when in contact with intracellular esterases, obtaining a fluorescent signal with internalized SPAchips.
Figure 1: CytoCHECK SPAchip® OHrad ROS Single-Detection Kit in Fibroblast 1095SK cell line. Images showing intracellular ROS SPAchip® in green, acquired with 40x magnification objective. In blue, nuclear staining by Hoechst 33342. In red, cytoplasmatic staining by Cell Mask™ Plasma Membrane. Scale bar: 20 µm.
Technical specifications
| Product code | S-003-ROSG |
| Amount | ~2.5 millions of SPAchips |
| Applications | Cell viability, proliferation, cell image acquisition |
| Assay Time | 30 minutes |
| Assay type | Living single-cell based |
| Solubility | Soluble in assay buffer (aqueous) |
| Analyte | ROS: hydroxyl radicals (•OH) |
| Detection method | Green fluorescence |
| Fluorescence | λex: 488 nm; λem: 520 nm |
| Compatible Platforms | Fluorescence microscopy, HCS/HCA platforms (20x magnification and over) and flow cytometry |
| Sample type | Adherent cells, suspension cells |
Image Acquisition and Quantitative Analysis
CytoCHECK SPAchip® kits are designed to dynamically shift fluorescence intensity values in response to intracellular changes in analyte concentration. Fluorescence intensity to quantify intracellular reactive oxygen species (ROS), by detecting hydroxyl radical dynamics inside live cells, is measured by exciting the live cell sample at 488 nm and detecting fluorescence emission at 520 nm. Data acquisition can be adapted to fluorescence microscopy and flow cytometry. Extra detection channels, such as transmitted light, are suggested to better identify and localize the cells and SPAchips of interest.
Additionally, a major advantage of CytoCHECK SPAchip® OHrad ROS Single-Detection Kit is that it is not sensitive to pH changes (Fig. 2), avoiding measurement artifacts due to pH variations in experimental setup.
Figure 2: CytoCHECK SPAchip® OHrad ROS Single-Detection Kit is not pH-sensitive. No significant effect of pH on the fluorescence of CytoCHECK OHrad ROS 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.
* Contact A4cell staff for assistance with imaging setup and data analysis. Ask for our cell biology image analysis guide at info@a4cell.com
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