Lipofuscin labeling through biorthogonal strain‐promoted azide‐alkyne cycloaddition for the detection of senescent cells

A new method for senescent cell detection is described, which is based on lipofuscin labeling with a fluorescent reporter through a biorthogonal strain‐promoted azide‐alkyne cycloaddition. The sensing protocol involves a first step where the interaction of lipofuscin with a Sudan Black B derivative containing an azide moiety (SBB‐N3) is carried out. In the final step, the azide moiety reacts with a fluorophore containing a cyclooctene ring (BODIPY). The efficacy of this two‐step protocol is assessed in senescent melanoma SK‐MEL‐103 cells, senescent triple‐negative breast cancer MDA‐MB‐231 cells and senescent WI‐38 fibroblasts. In all cases, a clear fluorescence pattern was observed in senescent cells, compared to proliferative cells, only when the SBB‐N3‐BODIPY probe was formed. Our results provide an alternative tool for the detection of senescent cells, based on an in situ bio‐orthogonal reaction for lipofuscin labeling.

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