Product name: IC60N
Live-cell imaging –based analysis of intracellular oxygen in single cells
The IC60N intracellular assays are based on the ability of O2 to quench the excited state of IC60N, a porphyrin-based, water-soluble, phosphorescent, oxygen-sensitive probe. The probe is passively loaded into cells by liposomal transfer or facilitated endocytosis. IC60N is compatible with fluorescence microscopes, confocal laser scanning microscopes, and other imaging systems. See product insert for assay principal.
There are two types of IC60N assays: one for suspension cells, and one for adherent cells:
1) IC60N-LCA: Live-cell imaging-based analysis of intracellular oxygen in eukaryotic cell suspensions.
Cell suspensions are pipetted onto the LiveCell® Array (LCA) from Molecular Cytomics, Boston, US. The LCA consists of 360 individual 8 µm deep wells which each trap a single suspension cell whilst maintaining cell viability. The LCA allows individual cells to be treated with various effectors and then analysed. The IC60N probe is loaded into cells and effectors added if required.
2) IC60N-ADH: Live cell imaging based analysis of intracellular oxygen in eukaryotic adherent cells.
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Note: High-throughput packages available on all products. Please contact us for more information.
LCA SLIDE*
The LCA slides are manufactured by Molecular Cytomics, Boston, MA, US. A single LCA* slide comprises 360, 8µm deep wells. Each one is capable of trapping a single suspension/primary cell without compromising viability, thereby allowing individual cells to be treated with various effectors and then analysed. The combination of MitoXpress™ IC60N with the Live Cell Array™ is the perfect solution for analyzing intracellular oxygen consumption in suspension and primary cells. For more information on the LCA slide, visit www.molecular-cytomics.com
A long distance LUCPFLN 40X objective from Olympus (or similar), is required only when using an LCA slide.
LOADING AGENT**
We have evaluated a range of loading techniques and reagents and found Endo-Porter, produced by GeneTools, LLC, to be the optimal loading agent for certain cell types, for example HepG2’s. For other cell types other loading reagents or techniques may be more appropriate. Endo-Porter contains low concentrations of DMSO that may trigger some undifferentiated cell types to differentiate. In this case, Gene Tools provide Aqueous Endo-Porter, although the aqueous formulation is considerably less effective than the DMSO formulation. Choose the loading agent or technique that works best for your cell type. For more information on Endo-Porter see www.gene-tools.com