Publikationsdatenbank

Protocol for in vivo fluorescence lifetime microendoscopic imaging of the murine femoral marrow (2025)

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Fiedler, Alexander F. (WE 2)

Niesner, Raluca A. (WE 2)

Quelle

Methods in microscopy

Bandzählung: 2

Heftzählung: 1

Seiten: 85 – 96

ISSN: 2942-3899

Sprache

Englisch

Verweise

URL (Volltext): https://www.degruyterbrill.com/document/doi/10.1515/mim-2024-0023/html

DOI: 10.1515/mim-2024-0023

Kontakt

Institut für Veterinär-Physiologie

Oertzenweg 19 b
14163 Berlin
+49 30 838 62600
physiologie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

We present a protocol for micro-endoscopic fluorescence lifetime imaging in the femoral marrow of mice allowing the analysis of NAD(P)H-dependent metabolism at single cell level, in vivo. Therefore, we employed a gradient refractive index (GRIN) lens system fixed to the mouse femur by a specialized implant. We provide step-by-step instructions for the practical usage of the microendoscopic femoral implant and discuss experimental parameters required for reliable NAD(P)H FLIM analysis, particularly referring to photon statistics and signal-to-noise ratio. Representative results indicate metabolic heterogeneity both in marrow tissue environment and among marrow LysM+ myeloid cells in vivo. We expect the here presented microendoscopic FLIM approach to enable the analysis of cellular functions and dysfunctions, beyond cellular metabolism, providing a better understanding of bone biology, in health and disease.