Cell aging - a relevant factor in live cell microscopy (mini-review)

Live-cell microscopy is gaining importance for studying cellular behavior in response to environmental cues. However, cell aging can result in modifications of various cellular structures and functions, affecting or distorting microscopy-based readouts. These changes include gene expression, nuclear architecture, energy metabolism, or changes in the mechanical properties of cell membranes and microtubules. In this mini-review, we briefly discuss how cell aging affects critical subcellular compartments and alters live-cell imaging outcomes. In contrast to many papers available on cell aging, here we are focusing on the influence of cell aging on the performance and outcome of advanced microscopy techniques such as super-resolution imaging, fluorescence lifetime imaging (FLIM), variable-angle total internal reflection fluorescence microscopy (VA-TIRFM), as well as micromanipulation techniques such as laser-assisted optoporation. Our findings highlight the importance of considering cell passage number and senescence markers in experimental design and data interpretation.