Shaping light for volumetric microscope imaging in the heart

用于心脏体积显微镜成像的整形光

• 批准号: EP/N029917/1
• 负责人: Jonathan Taylor
• 金额: $ 12.86万
• 依托单位: University of Glasgow
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FN029917%2F1
• 关键词: Shaping; light; volumetric; microscope; imaging

Recent advances in light sheet fluorescence microscopy have allowed biomedical researchers to watch and study living animals such as the zebrafish as they grow from a single cell to a fully functioning organism. However obtaining continuous 3D images presents a particular challenge in the heart (since it is constantly beating) and images become clouded and blurred as the animal grows larger and it is necessary to image through increasing amounts of overlying tissue to see the organ of interest.We will acquire microscope images using specific new illumination and imaging techniques we will develop, to allow us to obtain higher quality images than previously possible inside living tissue. In their raw form these images will not resemble conventional images, but with the help of the powerful imaging processing capabilities of modern computers, we will be able to analyze and combine the raw images to recover better images than would have otherwise been possible with conventional microscope imaging.Specifically, we will research and implement three techniques:1. Speckle light sheet imaging. Here instead of illuminating our sample with uniform light we will illuminate it with a random speckle field. Our raw images will therefore appear "dappled" and unclear, but following computer image processing the resultant images will be much sharper and less affected both by shadowing effects and by the overlying tissue that the light has passed through.2. Wavefront coding for focus-invariant synchronized heart imaging. When we take 3D video images of the heart, we have to cope with the fact that the heart is beating faster than we can normally obtain a complete 3D image of it. We overcome this by using image analysis and computer control to synchronize our image acquisition with the heartbeat. However this is particularly difficult because we would usually move the sample around in order to take the 3D image, and this spoils the synchronization. Wavefront coding lets us acquire images that no longer look like a clear image of the heart, but which remain the same as we move the sample, thus allowing us to build a much simpler and cheaper synchronized imaging system.3. Wavefront coding for snapshot volume imaging. Our synchronized imaging technique assumes that the heart is beating regularly, and by definition that will not be the case in many diseased hearts - which biologists are particularly interested in studying. We will overcome this problem by developing a method for extremely fast volume imaging. Normally the imaging speed is limited by how fast we can change the focus of our microscope, but wavefront coding will allow us to do the refocusing on a computer afterwards, thus allowing us to obtain 3D images much faster.These techniques together will offer new and improved methods for microscope imaging to look inside living animals, to help biologists better understand how the heart develops and functions - with the ultimate aim of improving medical treatments for human heart diseases.

光片荧光显微镜的最新进展使生物医学研究人员能够观察和研究活的动物，如斑马鱼，因为它们从一个单细胞成长为一个功能齐全的生物体。然而，获得连续3D图像在心脏中提出了特别的挑战（因为它是不断跳动的）和图像变得浑浊和模糊的动物越来越大，有必要通过增加覆盖组织的数量来看到感兴趣的器官。我们将使用特定的新照明和成像技术，我们将开发获取显微镜图像，使我们能够获得比以前更高质量的活体组织图像。这些原始图像与传统图像不同，但借助现代计算机强大的图像处理能力，我们将能够分析和联合收割机，恢复出比传统显微镜成像更好的图像。具体来说，我们将研究和实现三种技术：1.散斑光片成像。在这里，我们将用随机散斑场来照射样本，而不是用均匀的光来照射样本。因此，我们的原始图像会出现“斑点”和不清晰，但经过计算机图像处理后，所得到的图像将更加清晰，受阴影效应和光线穿过的覆盖组织的影响更小。2.用于焦点不变同步心脏成像的波前编码。当我们拍摄心脏的3D视频图像时，我们必须科普心脏跳动的速度比我们通常获得的完整3D图像快的事实。我们通过使用图像分析和计算机控制来克服这个问题，使我们的图像采集与心跳同步。然而，这是特别困难的，因为我们通常会移动样品来拍摄3D图像，这会破坏同步。波前编码让我们获得的图像不再看起来像心脏的清晰图像，而是在我们移动样本时保持不变，从而使我们能够构建一个更简单，更便宜的同步成像系统。3.用于快照卷成像的波前编码。我们的同步成像技术假设心脏有规律地跳动，根据定义，许多患病的心脏不会出现这种情况-生物学家对此特别感兴趣。我们将通过开发一种用于极快体积成像的方法来克服这个问题。通常，成像速度受到我们改变显微镜焦点的速度的限制，但波前编码将允许我们在计算机上进行重新聚焦，从而使我们能够更快地获得3D图像。这些技术将为显微镜成像提供新的和改进的方法，以观察活体动物的内部，帮助生物学家更好地了解心脏的发育和功能-最终目的是改善人类心脏病的医疗方法。