Wide-field microscope to support research programs in image cytometry and image-based cell separation

细胞术或细胞分析对生物学研究的许多领域至关重要。自20世纪60年代以来，传统流式细胞术一直是占主导地位的细胞术技术，因为它简单，高通量，并且可以与荧光激活的细胞分选相结合。然而，目前有许多样品和分析与流式细胞术不兼容。例如，(1)细胞样品可能无法承受流式细胞术的剪切应力；(2)细胞可以组织成多细胞簇，如球状体和类器官；(3)感兴趣的特征可能表现为某些标记的亚细胞定位。这些问题中的许多都可以通过显微镜成像来解决。然而，目前使用标准显微镜系统分析细胞的方法非常缓慢，并且不能提供分离特定细胞或集群以进行进一步分析的能力，例如通过繁殖和基因组测序。******本提案要求资助一个带有大画幅相机的宽视场显微镜，以填补几个多学科研究项目中对高通量成像的关键需求。该显微镜的主要特点是其扩大的视野，这大大增加了成像吞吐量，减少了图像采集时间。所要求的设备将支持五个动态研究项目，涵盖广泛的学科，包括开发图像细胞术的新算法和基于图像的细胞分离的新技术，器官芯片微流控系统的实时监测，基于图像的筛选来阐明血液凝固途径，高通量筛选来发现新的聚合物生物材料，利用多路显微镜研究细胞因子分泌的分子机制。******所要求的宽视场显微镜系统为研究生、本科生和博士后水平的HQP培训提供了一个多功能平台。学员将能够使用该平台快速获取大量显微镜数据，以调查特定的研究问题，开发图像细胞术的新算法，以及开发基于图像的细胞分离的新技术。在当前数据驱动的经济中，获取和分析大量数据的能力是许多科学和工程领域的宝贵技能。拟议的设备将为受训人员提供发展和展示这些技能的机会。*****

Cytometry, or the analysis of cells, is fundamentally important to many areas of biological research. Traditional flow cytometry has been the dominant cytometry technology since the 1960s because it is simple, high-throughput, and can be coupled with fluorescence activated cell sorting. However, there are many samples and analyses currently incompatible with flow cytometry. For example, (1) the cell sample may be unable to withstand the shear stress of flow cytometry; (2) cells may be organized into multicellular clusters, such as spheroids and organoids; (3) the features of interest may be presented as subcellular localization of certain markers. Many of these issues can be addressed using microscopy imaging. However, current methods for analyzing cells using standard microscopy systems are extremely slow and does not provide the ability to separate specific cells or clusters for further analysis, such as by propagation and genome sequencing.******This proposal requests funding for a wide-field microscope with a large-frame camera to fill a critical need for high-throughput imaging in several multi-disciplinary research programs. The key feature of this microscope is its enlarged field-of-view, which dramatically increases imaging throughput and reduces image acquisition times. The requested equipment will support five dynamic research programs spanning a wide range of disciplines, including the development of new algorithms for image cytometry and new technologies for image-based cell separation, real-time monitoring of organ-on-a-chip microfluidic systems, image based screening to elucidate blood coagulation pathways, high-throughput screening to discover new polymeric biomaterials, and multiplexed microscopy to elucidate the molecular mechanisms of cytokine secretion.******The requested wide-field microscopy system provides a versatile platform for HQP training at the graduate, undergraduate, and postdoctoral level. Trainees will be able to use this platform to rapidly acquire large amounts of microscopy data in order to investigate specific research questions, develop new algorithms for image cytometry, as well as to develop new technologies for image-based cell separation. In the current data-driven economy, the ability to acquire and analyze large amounts of data are valuable skills in many areas of science and engineering. The proposed equipment will provide trainees the opportunity to develop and demonstrate these skills.*****