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Much of the current understanding of thermal effects in biological systems is based on macroscopic measurements. There is little knowledge about the local thermostability or heat tolerance of subcellular components at the nanoscale. Herein, we show that gold nanorod-fluorescent nanodiamond (GNR-FND) hybrids are useful as a combined nanoheater/nanothermometer in living cells. With the use of a 594 nm laser for both heating and probing, we measure the temperature changes by recording the spectral shifts of the zero-phonon lines of negatively charged nitrogen-vacancy centers in FNDs. The technique allows us to determine the rupture temperatures of individual membrane nanotubes in human embryonic kidney cells, as well as to generate high temperature gradients on the cell membrane for photoporation and optically controlled hyperthermia. Our results demonstrate a new paradigm for hyperthermia research and application. [ABSTRACT FROM AUTHOR]