Generation of a temporal, spatial, and molecular map of in situ hematopoiesis

（请保持文字，不要PDF） 在此输入文本，它是应用程序的新摘要信息。此部分不得超过30行文本。 从发育和干细胞生物学的角度来看，造血系统可以说是研究得最好的组织。从历史上看，造血研究利用流式细胞术和细胞移植方法作为检查功能行为的金标准测定。虽然这些研究提供了一个优雅的造血教科书的观点，我们认为，我们的知识是如何早期血液形成工程在很大程度上是不完整的。这主要是由于利用传统的分析和读数的局限性，破坏了正常的骨髓组织结构，忽视了细胞的解剖定位。因此，在该领域的许多研究中缺少的一个关键生物组成部分是空间组织。基本的问题，如造血命运的承诺何时何地发生，什么确切的细胞中间体是干细胞和成熟的子细胞，以及什么身份和作用的细胞微环境（龛）是在造血过程中仍然在很大程度上未解决。在这里，我们提出了一个跨学科的方法，基于国家的最先进的活显微镜操作结合新的造血命运报告，使造血的全面研究完全在原位。使用新的两个光子引导收集直接从骨髓中的干细胞克隆，和纵向活成像分析的干细胞后代，我们的目标是产生一个空间，时间和分子地图造血单细胞分辨率。互补的邻近为基础的光标记方法将被用来编目的细胞组成的生态位在稳态，患病，和老年人的设置。通过从组织破坏性方法转向依赖于观察，操纵和分析骨髓内单个造血细胞的系统，我们希望获得对血液生产细胞组织的独特见解。

(PLEASE KEEP IN WORD, DO NOT PDF) Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. The hematopoietic system has been, arguably, the best well studied tissue from a developmental and stem cell biology perspective. Historically, the study of hematopoiesis has taken advantage of flow cytometry and cell transplantation approaches as the gold standard assays to examine functional behaviors. While these studies have provided an elegant textbook view of hematopoiesis, we argue here that our knowledge of how early blood formation works is largely incomplete. This is predominantly due to the limitations of utilizing traditional assays and readouts that disrupt the normal marrow tissue archicture and neglect the anatomical localization of cells. Thus, one key biological component missing in many studies in the field is that of spatial organization. Basic questions such as when and where hematopoietic fate commitment occurs, what the exact cellular intermediates are between a stem cell and mature daughter cells, and what the identity and role of the cellular microenvironement (niche) is in the hematopoietic process remain largely unsettled. Here, we propose an interdisciplinary approach based on state-of-the-art live microscopy manipulations in combination with novel hematopoietic fate reporters to enable a comprehensive study of hematopoiesis entirely in situ. Using novel two photon-guided collection of stem cell clones directly from the bone marrow, and longitudinal live-imaging analysis of stem cell progenies, we aim to generate a spatial, temporal and molecular map of hematopoiesis at single cell resolution. Complementary proximity-based photolabelling approaches will be taken to catalogue the cellular composition of the niche in the steady-sate, diseased, and aged settings. By moving away from tissue disruptive approaches to a system that relies on observation, manipulation, and analysis of single hematopoietic cells within the bone marrow, we hope to obtain unique insights into the cellular organization of blood production.