Abstract
SummaryMeasuring three-dimensional (3D) localization of hematopoietic stem cells (HSCs) within the bone marrow microenvironment using intravital microscopy is a rapidly expanding research theme. This approach holds the key to understanding the detail of HSC-niche interactions, which are critical for appropriate stem cell function. Due to the complex tissue architecture of the bone marrow and to the progressive introduction of scattering and signal loss at increasing imaging depths, there is no ready-made software to handle efficient segmentation and unbiased analysis of the data. To address this, we developed an automated image analysis tool that simplifies and standardizes the biological interpretation of 3D HSC microenvironment images. The algorithm identifies HSCs and measures their localization relative to surrounding osteoblast cells and bone collagen. We demonstrate here the effectiveness, consistency, and accuracy of the proposed approach compared to current manual analysis and its wider applicability to analyze other 3D bone marrow components.
Highlights
Precise regulation of somatic stem cell function is essential for the survival of multicellular living organisms ranging from C. elegans to humans
Measuring three-dimensional (3D) localization of hematopoietic stem cells (HSCs) within the bone marrow microenvironment using intravital microscopy is a rapidly expanding research theme. This approach holds the key to understanding the detail of HSC-niche interactions, which are critical for appropriate stem cell function
We developed an automated image analysis tool that simplifies and standardizes the biological interpretation of 3D HSC microenvironment images
Summary
Precise regulation of somatic stem cell function is essential for the survival of multicellular living organisms ranging from C. elegans to humans. The correct functioning of somatic stem cells depends on complex and dynamic interactions with specific cellular and molecular components of the microenvironment that surrounds them (together called ‘‘niche’’) (Scadden, 2014), and in vivo imaging of stem cells is an expanding and promising field that provides a unique perspective of their behavior in situ. To date, this approach has been directly responsible for generating new hypotheses on the crucial role of the stem cell niche (Ritsma et al, 2014; Rompolas et al, 2012). Others, have successfully used fluorescent dyes to label HSCs prior to transplantation to achieve their detection in vivo through the bone of anesthetized recipient mice (Kohler et al, 2009; Lo Celso et al, 2009)
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