Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers

Fluorescence thermometry is a microscopy technique in which fluorescent temperature sensor records changes as alterations of fluorescence signals. lifetime imaging (FLIM) promising method for quantitative analysis intracellular temperature. Recently, we developed small-molecule thermometers, termed Organelle Thermo Greens, that target various organelles and achieved mapping using FLIM. Despite its highly nature, FLIM-based cannot be used widely due to expensive instrumentation. Here, investigated the applicability limitations intensity (FI)-based analysis, more commonly than thermometry. Temperature gradients generated by artificial heat sources physiological produced brown adipocytes were visualized FI- By comparing two techniques, examined how shapes cells affect accuracy measurements. Based on results, concluded FI-based could “qualitative”, rather quantitative, under limited condition shape change dye leakage from organelle not critical.