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Publication : Optimized intravital three-photon imaging of intact mouse tibia links plasma cell motility to functional states.

First Author  Rakhymzhan A Year  2024
Journal  iScience Volume  27
Issue  10 Pages  110985
PubMed ID  39391739 Mgi Jnum  J:355220
Mgi Id  MGI:7738129 Doi  10.1016/j.isci.2024.110985
Citation  Rakhymzhan A, et al. (2024) Optimized intravital three-photon imaging of intact mouse tibia links plasma cell motility to functional states. iScience 27(10):110985
abstractText  Intravital deep bone marrow imaging is crucial to studying cellular dynamics and functions but remains challenging, and minimally invasive methods are needed. We employed a high pulse-energy 1650 nm laser to perform three-photon microscopy in vivo, reaching approximately 400 mum depth in intact mouse tibia. Repetition rates of 3 and 4 MHz allowed us to analyze motility patterns of fast and rare cells within unperturbed marrow and to identify a bi-modal migratory behavior for plasma cells. Third harmonic generation (THG) was identified as a label-free marker for cellular organelles, particularly endoplasmic reticulum, indicating protein synthesis capacity. We found a strong THG signal, suggesting high antibody secretion, in one-third of plasma cells while the rest showed low signals. We discovered an inverse relationship between migratory behavior and THG signal, linking motility to functional plasma cell states. This method may enhance our understanding of marrow microenvironment effects on cellular functions.
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