MarShie: a clearing protocol for 3D analysis of single cells throughout the bone marrow at subcellular resolution.

ABSTRACT: Analyzing immune cell interactions in the bone marrow is vital for understanding hematopoiesis and bone homeostasis. Three-dimensional analysis of the complete, intact bone marrow within the cortex of whole long bones remains a challenge, especially at subcellular resolution. We present a method that stabilizes the marrow and provides subcellular resolution of fluorescent signals throughout the murine femur, enabling identification and spatial characterization of hematopoietic and stromal cell subsets. By combining a pre-processing algorithm for stripe artifact removal with a machine-learning approach, we demonstrate reliable cell segmentation down to the deepest bone marrow regions. This reveals age-related changes in the marrow. It highlights the interaction between CX₃CR1

SUBMITTER: Mertens TF 

PROVIDER: S-EPMC10897183 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature