Confocal spectroscopy for cell 3D microthermometry

每两个人中就有一个会在有生之年被诊断出患有癌症，这对英国“更长时间保持健康”的目标构成了重大挑战。对于一些癌症，治疗创新提高了存活率，但对于其他癌症，如脑癌，结果在20年内几乎没有变化。癌症研究界越来越感兴趣的一个话题是新陈代谢在癌细胞行为中的关键作用。Warburg效应是癌细胞从氧化代谢到糖酵解代谢的一种代谢转换，已有数十年的文献记载，但对癌细胞代谢的性质和意义仍知之甚少。肿瘤细胞分泌的内源性致热物质在37-42℃的温度范围内会导致明显的高温。同时，细胞的不同部分可以处于不同的温度，线粒体比基础温度高出10℃以上。我们需要调查关于癌细胞代谢的基础知识，它在肿瘤生长中的作用，以及靶向肿瘤内更活跃的代谢区域进行治疗的可能性。值得注意的是，人们越来越意识到，这需要在完整的癌症组织中进行，在这种组织中，可以观察到癌细胞与细胞微环境的相互作用。癌细胞-微环境相互作用影响癌细胞生物学，并且不能有效地利用体外癌细胞培养来模拟。因此，至关重要的是，必须在组织切片培养中询问癌细胞的新陈代谢，并最终在啮齿动物模型中进行研究，为此，我们需要一种能够在厚组织中三维工作的成像技术，并能够提供癌症环境中的温度分布。这可以通过使用发光纳米颗粒作为探针来实现。这种纳米粒子可以比红细胞小500倍，当它们被某个波长的光(“颜色”)激发时，它们将重新发射不同波长的光。对这种重新发射的光的分析可以提供有关其环境温度的信息。重要的是，某些波长可以在组织中传播更长时间而不会衰减，这可以用于从内部区域获取信息。这将使温度地图的3D重建成为可能。

One in two people will be diagnosed with cancer during their lifetime, presenting a significant challenge to the UK goal of "staying healthy for longer". For some cancers, therapeutic innovations have increased survival, but for other cancers, such as brain cancer, outcomes have changed little in 20 years. A topic of increasing interest in the cancer research community is the critical role of metabolism in cancer cell behaviour. The Warburg effect, a metabolic switch from oxidative to glycolytic metabolism in cancer cells, has been documented for over decades, but much remains unknown about the nature and significance of cancer cell metabolism. The intrinsic pyrogenic substances secreted by tumour cells induce distinct hyperthermia in the temperature range of 37 to 42 C. Simultaneously, different parts of the cell can be at different temperatures, with mitochondria more than 10 C above basal temperature.We need to investigate fundamental unknowns about cancer cell metabolism, its role in cancer growth and the potential of targeting more metabolically active regions within cancer for therapy. Significantly, there is increasing awareness that this needs to be done in the context of intact cancer tissue, where the cancer cell interactions with the cellular microenvironment can be observed. Cancer cell-microenvironment interactions influence cancer cell biology and are not effectively modelled using in vitro cancer cell cultures. Crucially, then, cancer cell metabolism must be interrogated in tissue slice culture, and ultimately in rodent models, for which we need innovative technologies as proposed here.For this, it is necessary to have an imaging technique capable of working in three dimensions in thick tissue, and able to provide the temperature distributions in the cancer environment. This can be achieved by using luminescent nanoparticles as probes. Such nanoparticles can be 500 times smaller than a red blood cell, and when they are excited with light of a wavelength ("colour"), they will re-emit light in a different wavelength. The analysis of this re-emitted light can provide information about the temperature of its environment. Importantly, certain wavelengths can propagate longer in tissue without being attenuated, which can be used for obtaining information from inner areas. This will enable the 3D reconstruction of the map of temperatures.