Temperature
in mitochondria can be a critical indicator of cell
metabolism. Given the highly dynamic and inhomogeneous nature of mitochondria,
it remains a big challenge to quantitatively monitor the local temperature
changes during different cellular processes. To implement this task,
we extend our strategy on mitochondria-anchored thermometers from
“on–off” probe Mito-TEM to a ratiometric
probe Mito-TEM 2.0 based on the Förster resonance
energy transfer mechanism. Mito-TEM 2.0 exhibits not
only a sensitive response to temperature through the ratiometric changes
of dual emissions but also the specific immobilization in mitochondria
via covalent bonds. Both characters support accurate and reliable
detection of local temperature for a long time, even in malfunctioning
mitochondria. By applying Mito-TEM 2.0 in fluorescence
ratiometric imaging of cells and zebrafishes, we make a breakthrough
in the quantitative visualization of mitochondrial temperature rises
in different inflammation states.
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