Intravital Imaging of Bone Marrow Niches
Intravital Imaging of Bone Marrow Niches
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DOI：
10.1007/978-1-0716-1425-9_16
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发表时间：
2021-01-01
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影响因子：
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通讯作者：
Lo Celso, Cristina
中科院分区：
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作者：
Haltalli, Myriam L. R.;Lo Celso, Cristina
造血干细胞（HSC）有助于驱动成熟血细胞的生成，这对包括免疫防御和组织重塑在内的各种功能至关重要。它们存在于专门的骨髓（BM）微环境或生态位中，由细胞和化学成分组成，在调节长期HSC功能和存活中起关键作用。虽然流式细胞术方法大大推进了造血细胞的研究，使其能够在稳态和扰动情况下进行定量，但我们仍在了解支持不同谱系的特定BM微环境以及它们的小生境如何在压力和年龄下发生变化。在过去的十年中，成像技术的重大进展已经允许深入研究小鼠的HSC壁龛。在这里，我们描述了我们的协议可视化和分析的本地化，形态学和功能的小生境组件在小鼠颅骨，结合共聚焦和双光子活体显微镜，我们提出了测量血管通透性的具体例子。
Haematopoietic stem cells (HSCs) are instrumental in driving the generation of mature blood cells, essential for various functions including immune defense and tissue remodeling. They reside within a specialised bone marrow (BM) microenvironment, or niche, composed of cellular and chemical components that play key roles in regulating long-term HSC function and survival. While flow cytometry methods have significantly advanced studies of hematopoietic cells, enabling their quantification in steady-state and perturbed situations, we are still learning about the specific BM microenvironments that support distinct lineages and how their niches are altered under stress and with age. Major advances in imaging technology over the last decade have permitted in-depth studies of HSC niches in mice. Here, we describe our protocol for visualizing and analyzing the localization, morphology, and function of niche components in the mouse calvarium, using combined confocal and two-photon intravital microscopy, and we present the specific example of measuring vascular permeability.