{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T22:29:41Z","timestamp":1792794581231,"version":"4.5.0"},"reference-count":15,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T00:00:00Z","timestamp":1776124800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Michigan Integrative Musculoskeletal Health Core Center\u2019s (MiMHC) Structure, Composition, and Histology Core","award":["NIAMS P30 AR069620"],"award-info":[{"award-number":["NIAMS P30 AR069620"]}]},{"DOI":"10.13039\/100000090","name":"Department of Defense Congressionally Directed Medical Research Programs","doi-asserted-by":"crossref","award":["HT94252310327"],"award-info":[{"award-number":["HT94252310327"]}],"id":[{"id":"10.13039\/100000090","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) of the National Institutes of Health","award":["R01AR080035, UC2 AR082186"],"award-info":[{"award-number":["R01AR080035, UC2 AR082186"]}],"id":[{"id":"https:\/\/ror.org\/006zn3t30","id-type":"ROR","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000964","name":"Arthritis National Research Foundation","doi-asserted-by":"publisher","award":["1469667"],"award-info":[{"award-number":["1469667"]}],"id":[{"id":"10.13039\/100000964","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R00AR081894"],"award-info":[{"award-number":["R00AR081894"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01AR066028"],"award-info":[{"award-number":["R01AR066028"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["T32TR004371"],"award-info":[{"award-number":["T32TR004371"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["F31AR085942"],"award-info":[{"award-number":["F31AR085942"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Science Foundation Graduate Research Fellowship","award":["DGE 2241144"],"award-info":[{"award-number":["DGE 2241144"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,5,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Successful generation of high-quality spatial transcriptomics data from murine musculoskeletal tissues has been impeded by the challenge of preserving RNA integrity through the harsh tissue processing steps required for histological sectioning. In particular, the need to thoroughly fix and decalcify mineralized tissues has proven problematic. We detail a comprehensive sample processing pipeline for 3 common murine musculoskeletal tissue samples, enabling high-quality transcript detection via imaging-based spatial transcriptomics using the Xenium Prime 5K platform from 10x Genomics. Our protocol outlines methodological details for transcardiac perfusion, fixation, decalcification, paraffin processing, and a sample co-embedding strategy facilitating anatomically consistent and simultaneous sectioning of multiple samples onto the spatial transcriptomics slide. Rigorous quality control demonstrates high-quality tissue-specific outcomes across intact knee joints, tibiae, and lumbar spines from adult mice. Our pipeline enabled 70%-91% high-quality transcripts across synovium, meniscus, patellar tendon, articular cartilage, subchondral bone, cortical bone, bone marrow, muscle, fracture callus, and dorsal root ganglion (DRG) tissues. The average number of detected transcripts varied markedly between tissue types: soft tissues, such as synovium, patellar tendon, muscle, bone marrow, and callus exhibited ~200 to 400 transcript per cell; mineralized tissues, such as subchondral bone, meniscus, and cortical bone exhibited ~13 to 150 transcripts per cell; and highly active neuronal tissues, such as DRG neurons yielded 750-1100 transcripts per cell. Canonical cell markers within each tissue confirmed successful identification and representation of key cell types. Through rigorous sample quality assessment at multiple stages of processing, this protocol yields high-quality RNA transcript detection while preserving critical anatomical context and will serve as a valuable tool enabling spatial transcriptomic profiling of intact musculoskeletal tissue samples.<\/jats:p>","DOI":"10.1093\/jbmrpl\/ziag070","type":"journal-article","created":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T11:30:02Z","timestamp":1776166202000},"source":"Crossref","is-referenced-by-count":3,"title":["Spatial transcriptomic profiling of decalcified murine musculoskeletal samples via Xenium Prime 5K"],"prefix":"10.1093","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4126-8292","authenticated-orcid":false,"given":"Lindsey","family":"Lammlin","sequence":"first","affiliation":[{"name":"Department of Health Sciences and Technology, Institute for Biomechanics, ETH Z\u00fcrich , 8092 Z\u00fcrich ,","place":["Switzerland"]},{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huong X","family":"Tran","sequence":"additional","affiliation":[{"name":"Department of Health Sciences and Technology, Institute for Biomechanics, ETH Z\u00fcrich , 8092 Z\u00fcrich ,","place":["Switzerland"]},{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]},{"name":"Department of Biomedical Engineering, University of Michigan , Ann Arbor, MI 48109,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aanya","family":"Mohan","sequence":"additional","affiliation":[{"name":"Department of Health Sciences and Technology, Institute for Biomechanics, ETH Z\u00fcrich , 8092 Z\u00fcrich ,","place":["Switzerland"]},{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael D","family":"Newton","sequence":"additional","affiliation":[{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Scarlet C","family":"Howser","sequence":"additional","affiliation":[{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen J","family":"Redding","sequence":"additional","affiliation":[{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina A","family":"Capobianco","sequence":"additional","affiliation":[{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]},{"name":"Department of Biomedical Engineering, University of Michigan , Ann Arbor, MI 48109,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kurt D","family":"Hankenson","sequence":"additional","affiliation":[{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]},{"name":"Department of Biomedical Engineering, University of Michigan , Ann Arbor, MI 48109,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander J","family":"Knights","sequence":"additional","affiliation":[{"name":"Department of Orthopaedic Surgery, University of Michigan , Ann Arbor, MI 48109 ,","place":["United States"]},{"name":"Department of Orthopaedic Surgery, Washington University St. Louis , St. Louis, MO 63110 ,","place":["United States"]},{"name":"Center of Regenerative Medicine, Washington University St. Louis , St. Louis, MO 63117 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