title: Automated Identification and Localization of Hematopoietic Stem Cells in 3D Intravital Microscopy Data.
abstract: 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. 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.
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK. Electronic address: r.khorshed@imperial.ac.uk.
authors.forename: Reema A
authors.initials: RA
authors.lastname: Khorshed
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK.
authors.forename: Edwin D
authors.initials: ED
authors.lastname: Hawkins
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK.
authors.forename: Delfim
authors.initials: D
authors.lastname: Duarte
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK; Facility for Imaging by Light Microscopy, Imperial College London, London SW7 2AZ, UK.
authors.forename: Mark K
authors.initials: MK
authors.lastname: Scott
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK.
authors.forename: Olufolake A
authors.initials: OA
authors.lastname: Akinduro
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK.
authors.forename: Narges M
authors.initials: NM
authors.lastname: Rashidi
authors.affiliation: Facility for Imaging by Light Microscopy, Imperial College London, London SW7 2AZ, UK.
authors.forename: Martin
authors.initials: M
authors.lastname: Spitaler
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK. Electronic address: c.lo-celso@imperial.ac.uk.
authors.forename: Cristina
authors.initials: C
authors.lastname: Lo Celso
mesh_terms: D001853:Bone Marrow / Q000648:ultrastructure*; D006412:Hematopoietic Stem Cells / Q000648:ultrastructure*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D000069416:Intravital Microscopy; D055153:Stem Cell Niche