Incucyte® Cytolight Rapid Dye for Live-cell Cytoplasmic Labeling

Incucyte^® Cytolight Rapid Dye offers a fast, efficient method for the cytoplasmic labeling of live cells. Incucyte^® Cytolight Rapid Dyes are stable fluorescent reagents that freely pass through cell membranes; however, once inside, these dyes are transformed into cell-impermeant fluorescent probes that are transferred by dilution to daughter cells but not to adjacent cells within a population.

Addition of the Incucyte^® Cytolight Rapid Dyes at optimal concentrations to healthy cells is non-perturbing to cell growth or morphology and remains well-retained in cells. Validated for use with the Incucyte^® Live-Cell Analysis System and integrated software, Incucyte^® Cytolight Rapid Dyes are ideal for identifying and monitoring cells in mixed cultures up to 48-hours post labeling and can be used in conjunction with Incucyte^® Cell Health Reagents for simultaneous readouts of apoptosis or cytotoxicity.

Incucyte^® Cytolight Rapid Dyes are ideal for co-culture studies and can be used to identify and monitor subpopulations of cells for up to 48 hours following labeling. Please note the intensity of the Incucyte^® Cytolight Rapid signal will decrease over the course of the assay as it is diluted between daughter cells. For this reason, measurements beyond 48 hours post-labeling are generally not recommended. These reagents are also not recommended for measuring proliferation in fast growing cells over a long period of time as the rapid decline in fluorescence, due to reagent dilution between daughter cells, can impair fluorescence area and count metrics.

Presentation, Storage and Stability

Incucyte^® Cytolight Rapid Dyes are supplied as dry powders in quantities capable of labeling between 10 and 90 million cells depending on optimized concentration. The dry powder should be stored desiccated at -20° C. Once solubilized with DMSO, the solution should be stored at -20° C and protected from light. When stored as described, the dry powder will be stable for one year from date of receipt and the reconstituted solution is stable for at least one month.

Recommended Use

Solubilize Incucyte^® Cytolight Rapid Dyes with high-quality DMSO to prepare stock solutions as listed in the Incucyte^® Cytolight Rapid Product Guide in the Documents tab. The reagents may then be diluted in phosphate-buffered saline for labeling cells.

Labeling concentrations for the reagents will need to be optimized for each cell type to avoid toxicity and to ensure that the fluorescent signal remains detectable for the duration of your experiment. We recommend making this assessment by testing a range of concentrations of the dye and using Incucyte^® integrated analysis software confluence and fluorescence metrics to identify the optimal concentration that yields a sufficient signal with minimal cell growth perturbation within your experiment. The working concentration range for each dye can be found in the Incucyte^® Cytolight Rapid Product Guide.

When used with an Incucyte^® Live-Cell Analysis System, we recommend data collection every 1–3 hours.