Optimized intravital three-photon imaging of intact mouse tibia links plasma cell motility to functional states
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 <i>in vivo</i>, reaching ≈400 μm 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.
Authors
- Robert Günther (ORCID: https://orcid.org/0000-0002-3533-9747)
- Volker Andresen (ORCID: https://orcid.org/0000-0002-8960-6599)
- Jay Roodselaar (ORCID: https://orcid.org/0000-0002-9321-7836)
- Laura Wooldridge
- Asylkhan Rakhymzhan (ORCID: https://orcid.org/0000-0003-3152-1557)
- Alexander F. Fiedler (ORCID: https://orcid.org/0000-0003-3258-5640)
- Astrid Haibel
- Carolin Ulbricht (ORCID: https://orcid.org/0000-0003-2983-6242)
- Judith Heidelin
- Ruth Leben (ORCID: https://orcid.org/0000-0001-8188-2890)
- Ralf Köhler (ORCID: https://orcid.org/0000-0001-9563-2913)
- Georg N. Duda (ORCID: https://orcid.org/0000-0001-7605-3908)
- Anja E. Hauser (ORCID: https://orcid.org/0000-0002-7725-9526)
- Raluca Niesner (ORCID: https://orcid.org/0000-0001-8969-5432)
- Anne Bias (ORCID: https://orcid.org/0009-0002-4242-0946)
- Ingeborg Beckers
- Scott R. Domingue
- Yu Cao (ORCID: https://orcid.org/0009-0008-9631-2450)
Institutions
- Berliner Hochschule für Technik (DE)
- Thorlabs (Germany) (DE)
- Miltenyi Biotec (Germany) (DE)
- Bielefeld University (DE)
- Leibniz Association (DE)
- Humboldt-Universität zu Berlin (DE)
- German Rheumatism Research Centre (DE)
- Thorlabs (United States) (US)
- Freie Universität Berlin (DE)
Publication Details
- Journal
- iScience
- Published
- 2024-09-17
- DOI
- https://doi.org/10.1016/j.isci.2024.110985
- Citations
- 6
- Primary Topic
- Erythrocyte Function and Pathophysiology
- Type
- article
- Field-Weighted Citation Impact
- 1.96
Funders
- Deutsche Forschungsgemeinschaft
- Helmholtz-Zentrum Berlin für Materialien und Energie