Leveraging the spectral sorting revolution to gain deeper insights into cellular and tissue biology

人类和其他复杂的生物体是由数十亿个称为细胞的独立构件组成的。虽然这些细胞有一些共同的特征，但它们也往往是高度专门化的，从组成我们大脑的神经元，到将氧气从我们的肺部输送到身体其他部位的红细胞，再到排列在我们肠道并从我们所吃的食物中吸收营养的上皮细胞。了解这些细胞是如何发展这种特殊功能的，是生物学研究面临的一个基本问题，有助于我们理解生命本身的基本规则。科学家用来了解细胞如何发育和功能的一项技术是根据某些特征分离它们，特别是它们制造的特殊蛋白质，这些蛋白质可能会因细胞类型的不同而显著不同。细胞内的不同蛋白质可以使用带有特定荧光颜色(例如红色、蓝色或绿色)的蛋白质或染料进行标记。然后，一种被称为细胞分选器的专门设备可以使用探测器来读取每个细胞产生的荧光量，并根据细胞所拥有的荧光信号的组合将每个细胞物理地分类到不同的试管中。通过这种方式，细胞分选器可以将不同细胞类型的复杂混合物分成组，以便科学家可以单独研究每种细胞类型。我们正在申请资金，将在剑桥医学研究所(CIMR)现有的核心设施中安装一个新的细胞分选器，在那里，CIMR、剑桥干细胞研究所和剑桥大学及其周边地区的研究人员都可以使用它。核心设施非常繁忙，两个应用研究所的50个研究团队中的许多学生和研究人员都在使用它。我们必须购买一台新的分拣机，因为我们目前的分拣设备正在老化并迅速过时，实际上两年后制造商将不再为其提供支持。然而，尽管我们必须获得一个替换的分选器，但我们要求的特定设备--大脚光谱分选器--远远不止是一个简单的同类替换。由于过去几年分拣技术的重大变化，这款分拣机能够检测到比我们目前即将报废的设备更多的荧光颜色。这意味着研究人员将能够利用它来进行更复杂的实验，同时跟踪许多蛋白质，并更详细地了解细胞的功能和发育。分选速度也更快，最高可达7万个细胞/秒。这将节省资金，因为可以在相同的时间内对更多的单元格进行排序，而且它还将使数据变得更好，因为可以基于更大数量的单元格进行比较。它对细胞的分选也更温和，这使得它适合于特别脆弱和困难的细胞类型，例如我们血液中所有不同类型的细胞都来自的干细胞。总体而言，这是对我们现有设备的一次非常重要的升级，并开启了许多令人兴奋的研究机会。任何想要使用它的研究人员，包括剑桥以外的研究所和大学的研究人员，以及公司的用户，都可以使用新的分选器。所有用户都将为使用这台机器付费，这将有助于我们维护它并使其以最佳状态运行多年。它将由一支高技能和经验丰富的团队管理和运营，他们已经运营CIMR Flow核心设施10年。这些杰出的科学家与研究团队密切合作，帮助他们设计和提供实验，这是一种合作和支持的方法，使科学变得更好。大学里没有类似的机器，它的获得将使研究人员能够进行全新类型的实验，以了解细胞如何功能和发育的基本问题。

Humans and other complex organisms are made up of billions of individual building blocks called cells. While these cells share certain common features, they are also often highly specialised, from the neurons that make up our brain, to the red blood cells that carry oxygen from our lungs to the rest of our body, to the epithelial cells that line our intestine and absorb nutrients from the food we eat. Understanding how these cells develop such specialised functions is a fundamental question facing biological research, helping us to understand the essential rules of life itself. One technology that scientists use to understand how cells develop and function is to separate them based on certain characteristics, particularly the specialised proteins that they make, which can differ significantly from one cell type to another. Different proteins within cells can be 'tagged' using proteins or dyes with specific fluorescent colours (e.g. red, blue or green). A piece of specialised equipment known as a cell sorter can then use detectors to 'read' the amount of fluorescence each cell produces, and physically sort each cell into a different tube depending on the combination of fluorescent signals it has. In this way cell sorters can separate a complex mixture of different cell types into groups so that scientists can study each cell type separately.We are requesting funding for a new cell sorter to be housed in an already existing core facility in the Cambridge Institute for Medical Research (CIMR), where it will be accessed by researchers in CIMR, the Cambridge Stem Cell Institute and across the University of Cambridge and surrounding area. The core facility is very busy and is used by students and researchers in many of the >50 research teams in the two applying institutes. It is critical we that purchase a new sorter because our current sorting equipment is ageing and rapidly becoming obsolete, and in fact will no longer be supported by the manufacturer in two years time. However while it is essential that we obtain a replacement sorter, the specific equipment we are requesting, the Bigfoot Spectral Sorter, is much more than a simple like-for-like replacement. Because of significant changes in sorting technology in the past few years, this sorter is able to detect many more fluorescent colours than our current near end-of-life equipment. This means that researchers will be able to use it to carry out more complex experiments that track many proteins at the same time, and understand cell function and development in much greater detail. The sorting speed is also faster, up to 70,000 cells/second. This will save money as more cells can be sorted in the same amount of time, and it will also make the data even better because comparisons can be based on much larger numbers of cells. It is also gentler to the cells as it sorts, which makes it suitable for particularly fragile and difficult cell types, such as the stem cells from which all the different kinds of cells in our blood all originate. Overall, this is a very significant upgrade on our current equipment and opens up many exciting research opportunities.The new sorter will be available to any researcher who wants to use it, including those from institutes and universities outside Cambridge, as well as users from companies. All users will pay for using the machine, which will help us maintain it and keep it running optimally for many years. It will be managed and run by a highly skilled and experienced team who have been running the CIMR Flow Core Facility for >10 years. These outstanding scientists work very closely with the research teams to help them design and deliver their experiments, a collaborative and supportive approach that makes the science better. There is no similar machine within the University, and its acquisition will allow researchers to carry out completely new kinds of experiments to understand fundamental questions of how cells function and develop.