Enhancing biochemical resolution by hyper-dimensional imaging microscopy

Enhancing biochemical resolution by hyper-dimensional imaging microscopy
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通过超维成像显微镜提高生化分辨率

DOI:
10.1101/431452
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发表时间:
2018
期刊:
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影响因子:
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通讯作者:
Esposito A
Esposito A
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作者:
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二十年来的快速创新提高了荧光显微镜的空间分辨率,使分子分辨率具有低侵入性和高特异性。荧光显微镜还使科学家和临床医生能够绘制和定量生物样品的物理化学性质(例如,分析物浓度,酶活性和蛋白质-蛋白质相互作用)。但荧光显微镜的生化分辨能力不如其空间分辨能力优化。目前的技术通常只观察荧光的个别性质,从而限制了传感和多路复用的机会。在这里,我们展示了一种新的,据我们所知,成像范式,超维成像显微镜,它可以同时有效地量化生物样品中荧光发射的所有特性(激发态寿命,极化和光谱),超越了现有的限制。这种荧光特征的同时检测最大限度地提高了荧光显微镜的生化分辨能力,从而提供了增强传感能力和实现大量多路分析的手段。正如质谱中的多维分离和核磁共振中的多维光谱赋予了蛋白质组学和结构生物学力量一样,我们设想前所未有的多维成像显微镜光谱将催化系统生物学和医学诊断的进步。
Two decades of fast-paced innovation have improved the spatial resolution of fluorescence microscopy to enable molecular resolution with low invasiveness and high specificity. Fluorescence microscopy also enables scientists and clinicians to map and quantitate the physicochemical properties (e.g., analyte concentration, enzymatic activities, and protein-protein interactions) of biological samples. But the biochemical resolving power of fluorescence microscopy is not as well optimized as its spatial resolution. Current techniques typically observe only the individual properties of fluorescence, thus limiting the opportunities for sensing and multiplexing. Here, we demonstrate a new, to our knowledge, imaging paradigm, hyperdimensional imaging microscopy, which quantifies simultaneously and efficiently all the properties of fluorescence emission (excited-state lifetime, polarization, and spectra) in biological samples, transcending existing limitations. Such simultaneous detection of fluorescence features maximizes the biochemical resolving power of fluorescence microscopy, thereby providing the means to enhance sensing capabilities and enable heavily multiplexed assays. Just as multidimensional separation in mass-spectroscopy and multidimensional spectra in NMR have empowered proteomics and structural biology, we envisage that hyperdimensional imaging microscopy spectra of unprecedented dimensionality will catalyze advances in systems biology and medical diagnostics.
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