A critique of methods for temperature imaging in single cells

Baffou, Guillaume; Rigneault, Hervé; Marguet, Didier; Jullien, Ludovic

doi:10.1038/nmeth.3073

Cited by 214 publications

(227 citation statements)

References 29 publications

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“…The question of temperature heterogeneity and the interpretation of single cell thermometry data is the subject of on-going debate in the field of cellular thermosensing 58–61 , and a recent commentary proposed that temperature differentials measurable in cells should not exceed a μK range 58 . As such, the apparent temperature increase from 25 °C to 42 °C in WT-1 cells following isoproterenol stimulation as measured by ERthermAC staining would appear, at first sight, surprising (assuming the initial temperature in the ER is equivalent to that of the culture medium at 25 °C).…”

Section: Discussionmentioning

confidence: 99%

Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes

Kriszt

Arai

Itoh

et al. 2017

Sci Rep

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The identification of brown adipose deposits in adults has led to significant interest in targeting this metabolically active tissue for treatment of obesity and diabetes. Improved methods for the direct measurement of heat production as the signature function of brown adipocytes (BAs), particularly at the single cell level, would be of substantial benefit to these ongoing efforts. Here, we report the first application of a small molecule-type thermosensitive fluorescent dye, ERthermAC, to monitor thermogenesis in BAs derived from murine brown fat precursors and in human brown fat cells differentiated from human neck brown preadipocytes. ERthermAC accumulated in the endoplasmic reticulum of BAs and displayed a marked change in fluorescence intensity in response to adrenergic stimulation of cells, which corresponded to temperature change. ERthermAC fluorescence intensity profiles were congruent with mitochondrial depolarisation events visualised by the JC-1 probe. Moreover, the averaged fluorescence intensity changes across a population of cells correlated well with dynamic changes such as thermal power, oxygen consumption, and extracellular acidification rates. These findings suggest ERthermAC as a promising new tool for studying thermogenic function in brown adipocytes of both murine and human origins.

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Section: Discussionmentioning

confidence: 99%

Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes

Kriszt

Arai

Itoh

et al. 2017

Sci Rep

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“…Such higher mass of internalised heating agents leads theoretically to higher elevation of temperature, although a real temperature increase inside the cells induced by MFH is still a subject of debate in the community, due to their too small size for the heating power to compensate the dominant thermal losses into the surrounding environment. 37 Non-thermal effects are also hypothesised to contribute to the cytotoxicity in cellular MFH, like for example a direct mechanical destabilisation of the cell membranes (the plasma membrane during IONP internalisation and later the lysosome membrane): Also called "nanoflowers" as ascribed to their morphology, multicore IONPs exhibit a pronounced surface roughness that might cause membrane deformation and tearing, when comparing the high magnification TEM view of lysosomes loaded with multicore IONPs (Fig. S23) to the micrograph of a lysosome filled with monocore IONPs (Fig.…”

Section: Molecular Systems Design and Engineering Papermentioning

confidence: 99%

Monocorevs.multicore magnetic iron oxide nanoparticles: uptake by glioblastoma cells and efficiency for magnetic hyperthermia

Hemery

Genevois

Couillaud

et al. 2017

Mol. Syst. Des. Eng.

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“…Indeed, the validity of subcellular temperature changes reported in various papers has been challenged 34, 35 . Baffou et al .…”

Section: Introductionmentioning

confidence: 99%

Measurement of local temperature increments induced by cultured HepG2 cells with micro-thermocouples in a thermally stabilized system

Yang

et al. 2017

Sci Rep

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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.

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A critique of methods for temperature imaging in single cells

Cited by 214 publications

References 29 publications

Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes

Optical visualisation of thermogenesis in stimulated single-cell brown adipocytes

Monocorevs.multicore magnetic iron oxide nanoparticles: uptake by glioblastoma cells and efficiency for magnetic hyperthermia

Measurement of local temperature increments induced by cultured HepG2 cells with micro-thermocouples in a thermally stabilized system

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