Removing physiological motion from intravital and clinical functional imaging data.
Removing physiological motion from intravital and clinical functional imaging data.
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DOI：
10.7554/elife.35800
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发表时间：
2018-07-09
期刊：
影响因子：
7.7
通讯作者：
Timpson P
中科院分区：
文献类型：
--
作者：
Warren SC;Nobis M;Magenau A;Mohammed YH;Herrmann D;Moran I;Vennin C;Conway JR;Mélénec P;Cox TR;Wang Y;Morton JP;Welch HC;Strathdee D;Anderson KI;Phan TG;Roberts MS;Timpson P
活体显微镜可以提供独特的见解，生物过程中的功能，在一个本地的情况。然而，由呼吸、呼吸和心跳引起的生理运动可能带来重大挑战，特别是对于诸如荧光寿命成像（FLIM）的功能读数，其需要更长的采集时间来获得定量读数。在这里，我们提出和基准Galene，一个多功能的多平台软件工具，用于基于图像的校正样品运动模糊的时间分辨和传统的激光扫描荧光显微镜数据在二维和三维。我们表明，Galene是能够解决活体FLIM-FRET图像的腹腔内器官在小鼠模型和人体皮肤组织成像受到广泛的生理运动的NADH自体荧光。因此，Galene可以在稳定的成像平台并不总是可能的情况下实现FLIM成像，并挽救先前丢弃的定量成像数据。了解分子和细胞在活体动物中的行为可以让研究人员深入了解癌症等疾病的问题，以及这些疾病的潜在治疗方法的效果。一些工具可以帮助我们了解这些过程。例如，荧光“生物传感器”改变颜色，告诉我们特定蛋白质的活性。这可以表明药物在肿瘤的不同部位的效果如何。高分辨率显微镜可以对单个细胞甚至细胞的特定部分中发生的事件进行成像。然而，由于心跳或呼吸引起的微小运动会使图像变得模糊，从而难以研究活体动物。这对于需要几分钟才能捕获的图像来说尤其成问题。Warren等人现在已经开发了一种新的开源软件工具，称为Galene。该工具可以校正通过称为荧光寿命成像显微镜（FLIM）的技术收集的图像中的微小移动。因此，可以在以前不可能的情况下捕获清晰的图像。例如，Warren等人观察了癌细胞在24小时内从脾脏迁移到小鼠肝脏的过程，并使用荧光生物传感器表明，重新使用的药物会干扰细胞附着在肝脏上的程度。此外，Warren等人使用该软件在志愿者手臂上拍摄了人体皮肤的稳定3D图像，可用于研究药物渗透。Galene可以帮助研究人员研究活体动物的广泛生物过程。该软件还可以应用于现有的数据，以清理模糊的图像。在未来，Galene可以进一步发展，以配合手术中使用的成像技术。例如，外科医生可以用它来帮助他们找到肿瘤的边缘。
Intravital microscopy can provide unique insights into the function of biological processes in a native context. However, physiological motion caused by peristalsis, respiration and the heartbeat can present a significant challenge, particularly for functional readouts such as fluorescence lifetime imaging (FLIM), which require longer acquisition times to obtain a quantitative readout. Here, we present and benchmark Galene, a versatile multi-platform software tool for image-based correction of sample motion blurring in both time resolved and conventional laser scanning fluorescence microscopy data in two and three dimensions. We show that Galene is able to resolve intravital FLIM-FRET images of intra-abdominal organs in murine models and NADH autofluorescence of human dermal tissue imaging subject to a wide range of physiological motions. Thus, Galene can enable FLIM imaging in situations where a stable imaging platform is not always possible and rescue previously discarded quantitative imaging data. Understanding how molecules and cells behave in living animals can give researchers key insights into what goes wrong in diseases such as cancer, and how well potential treatments for these diseases work. A number of tools help us to see these processes. For example, fluorescent ‘biosensors’ change colour to tell us how active a particular protein is. This can indicate how well a drug works in different parts of a tumour. High resolution microscopy makes it possible to image events happening in single cells, or even specific parts of a cell. However, small movements like those due to the heartbeat or breathing can blur the images, making it difficult to study living animals. This is particularly problematic for images that take several minutes to capture. Warren et al. have now developed a new open source software tool called Galene. The tool can correct for small movements in images collected by a technique called fluorescence lifetime imaging microscopy (FLIM). As a result, clear images can be captured in situations that were not previously possible. For example, Warren et al. watched cancer cells migrating to the liver of a mouse from the spleen over 24 hours, and, using a fluorescent biosensor, showed that a repurposed drug interferes with how well the cells can attach to the liver. In addition, Warren et al. used the software to take steady 3D images of human skin in a volunteer’s arm, which could be used to study drug penetration. Galene could help researchers to study a wide range of biological processes in living animals. The software can also be applied to existing data to clean up blurred images. In the future Galene could be further developed to work with the imaging techniques used during surgery. For example, surgeons could use it to help them find the edges of tumours.