Lattice Light Sheet Microscopy for the Biosciences

为了了解细胞和生物体的功能，研究人员使用了各种显微镜来“观察”它们的内部。通常，我们将使用荧光标记来标记细胞内的蛋白质，然后使用荧光显微镜来观察蛋白质是如何组织的，以及这可能会随着时间的推移而变化。这对于构筑我们对蛋白质功能的理解非常重要。大多数显微镜向整个细胞(或生物体)照射荧光灯，但只对其中的一小部分成像。不幸的是，细胞对光很敏感，这种方法经常在我们观察时损害细胞，当我们试图对细胞成像时，细胞可能会死亡。为了克服这个问题，我们计划使用一种新型的显微镜，它只在一个狭窄的平面上照射细胞(或生物体)，我们只成像这个照亮的区域。与其他类型的显微镜相比，这种方法大大减少了细胞经历的光量，对细胞的损害要小得多，并且允许我们在不影响细胞行为的情况下对活细胞进行更长时间的成像。这种类型的光片显微镜产生的光平面非常薄(约半微米)，比细胞(约20微米厚)薄得多，这不仅减少了细胞所经历的光量，还帮助我们以高细节对它们进行成像。这种新的显微镜还帮助我们快速成像，每秒约50次，因此可以捕捉到行为的快速变化。用来产生这个薄光平面的物镜和用来对其中的荧光成像的物镜都放在样品的下面，这使得我们可以很容易地在培养皿中培养细胞，并将它们直接放在显微镜上成像。总体而言，这种新的显微镜将帮助我们了解特定的蛋白质如何在有丝分裂中发挥作用，以及如何推动癌症等疾病，研究细胞如何分化为肌肉和其他形式的组织，如使用细胞球(细胞器)，了解植物如何感知重力，感觉神经元中的信号，细胞如何分泌其内容物，以及生物体如何感知压力。它将支持利兹大学和约克郡地区广泛的额外研究。

To understand how cells and organisms function, researchers use a wide range of microscopes to 'see' inside them. Typically, we will use a fluorescent marker to label a protein inside a cell, and then use a fluorescent microscope to see how the protein is organised, and how this might change with time. This is really important in framing our ideas about how a protein functions. Most microscopes shine fluorescent light on the entire cell (or organism) but only image a small part of it. Unfortunately, cells are sensitive to light, and this approach often damages the cell as we look, and cells can die as we try to image them. To overcome this problem, we plan to use a new type of microscope, which only shines light in a single narrow plane across the cell (or organism), and we only image this illuminate region. This approach greatly reduces the amount of light that the cell experiences, which is much less damaging to the cell, and allows us to image living cells for much longer without affecting their behaviour, compared to other kinds of microscopes. The plane of light that this type of 'light sheet' microscope generates is very thin (about half a micron), much thinner than cells (which are about 20 microns thick), which not only reduces the amount of light that the cell experiences, but also helps us to image them with a high amount of detail. This new microscope also helps us to image rapidly, at about 50 times a second, so that rapid changes in behaviour are captured. The objective used to generate this thin plane of light and the objective used to image the fluorescence within it are both placed below the sample, which makes if easy for us to grow cells in dishes and place them straight onto the microscope to image them. Overall, this new microscope will help us see how specific proteins play a role in mitosis, and drive diseases such as cancer, investigate how cells differentiate into muscle, and into other forms of tissue, such as the retina, using balls of cells (organelles), understand how plants sense gravity, signalling in sensory neurones, how cells secrete their contents and how organisms sense stress. It will support a wide range of additional research across the University of Leeds, and in the Yorkshire region.