Protocol for isotype-resolved 3D imaging and quantification of bone marrow plasma cells in murine femurs

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Abdouramane Camara Tony Chen Anqi Xu Dinis Calado
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Abstract

Bone marrow plasma cells maintain durable antibody responses, but their tissue localization and morphology are difficult to quantify in situ. Here, we present a protocol for isotype-resolved 3D imaging and quantification of bone marrow plasma cells in murine femurs. We describe steps for fixation, optimal cutting temperature (OCT) coumpound embedding, longitudinal opening of mouse femurs, whole-mount immunostaining, optical clearing, and confocal 2D/3D imaging. We detail an Imaris-based workflow for reproducible single-cell segmentation, isotype-resolved counting, and the extraction of morphological features from defined regions of interest.

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Journal STAR Protocols
Volume 7
Issue number 3
Pages 104775
Available online
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Publisher website (DOI) 10.1016/j.xpro.2026.104775
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Europe PubMed Central 42579555
Pubmed 42579555

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