To monitor the temperature distribution of a cell and its changes under varied conditions is currently a technical challenge. A variety of non-contact methods used for measuring cellular temperature have been developed, where changes of local temperature at cell-level and sub-cell-level are indirectly calculated through the changes in intensity, band-shape, bandwidth, lifetime or polarization anisotropy of the fluorescence spectra recorded from the nano-sized fluorescent materials pre-injected into the target cell. Unfortunately, the optical properties of the fluorescent nano-materials may be affected by complicated intracellular environment, leading to unexpected measurement errors and controversial arguments. Here, we attempted to offer an alternative approach for measuring the absolute increments of local temperature in micro-Testing Zones induced by live cells. In this method, built-in high-performance micro-thermocouple arrays and double-stabilized system with a stability of 10 mK were applied. Increments of local temperature close to adherent human hepatoblastoma (HepG2) cells were continuously recorded for days without stimulus, showing frequent fluctuations within 60 mK and a maximum increment by 285 mK. This method may open a door for real-time recording of the absolute local temperature increments of individual cells, therefore offering valuable information for cell biology and clinical therapy in the field of cancer research.
In this paper, the authors introduced a novel method for the fabrication of microdevices on freestanding Si3N4 thin-film windows with a single-side mask aligner. A thin-film microdevice located on the front surface of such a freestanding window was demonstrated, where the backside of Si wafer was performed with KOH wet etching. The results showed that an alignment error could be controlled in condition less than 20 μm. By using the freestanding device, a weak change in local temperature under an e-beam in a scanning electron microscope (FEI QUANTA 600F) was detected with better sensitivity.
Monitoring the thermal responses of individual cells to external stimuli is essential for studies of cell metabolism, organelle function, and drug screening. Fluorescent temperature probes are usually employed to measure the temperatures of individual cells; however, they have some unavoidable problems, such as, poor stability caused by their sensitivity to the chemical composition of the solution and the limitation in their measurement time due to the short fluorescence lifetime. Here, we demonstrate a stable, non-interventional, and high-precision temperature-measurement chip that can monitor the temperature fluctuations of individual cells subject to external stimuli and over a normal cell life cycle as long as several days. To improve the temperature resolution, we designed temperature sensors made of Pd–Cr thin-film thermocouples, a freestanding Si3N4 platform, and a dual-temperature control system. Our experimental results confirm the feasibility of using this cellular temperature-measurement chip to detect local temperature fluctuations of individual cells that are 0.3–1.5 K higher than the ambient temperature for HeLa cells in different proliferation cycles. In the future, we plan to integrate this chip with other single-cell technologies and apply it to research related to cellular heat-stress response.
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