Greiner Stella, Ebrahimi Mahyasadat, Rodewald Marko, Urbanek Annett, Meyer-Zedler Tobias, Schmitt Michael, Neugebauer Ute, Popp Jürgen
Leibniz Institute of Photonic Technology (Leibniz IPHT), Member of the Research Alliance "Leibniz Health Technologies", Member of the Leibniz Center for Photonics in Infection Research (LPI) Jena, Albert-Einstein-Straße 9, 07745 Jena, Germany.
Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Helmholtzweg 4, 07743 Jena, Germany.
Bioengineering (Basel). 2025 Feb 10;12(2):170. doi: 10.3390/bioengineering12020170.
In this study, thin mouse kidney sections from healthy mice and those infected leading to acute and chronic sepsis were examined with two-photon excited fluorescence lifetime imaging (2P-FLIM) using the endogenous fluorescent coenzymes nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD). The results presented show that this approach is a powerful tool for investigating cell metabolism in thin tissue sections. An adapted measurement routine was established for these samples by performing a spectral scan, identifying a combination of two excitation wavelengths and two detection ranges suitable for detailed scan images of NADH and FAD. Selected positions in thin slices of the renal cortex of nine mice (three healthy, three with chronic sepsis, and three with acute sepsis) were studied using 2P-FLIM. In addition, overview images were obtained using two-photon excited fluorescence (2PEF) intensity. This study shows that healthy kidney slices differ considerably from those with acute sepsis with regard to their fluorescence lifetime signatures. The latter shows a difference in metabolism between the inner and outer cortex, indicating that outer cortical tubular cells switch their metabolism from oxidative phosphorylation to glycolysis in kidneys from mice with acute sepsis and back in later stages, as seen for mice with chronic infections. These findings suggest that 2P-FLIM could serve as a powerful tool for early-stage sepsis diagnosis and monitoring metabolic recovery during treatment.
在本研究中，使用内源性荧光辅酶烟酰胺腺嘌呤二核苷酸（NADH）和黄素腺嘌呤二核苷酸（FAD），通过双光子激发荧光寿命成像（2P-FLIM）对来自健康小鼠以及感染后导致急性和慢性败血症的小鼠的薄肾切片进行了检查。呈现的结果表明，这种方法是研究薄组织切片中细胞代谢的有力工具。通过执行光谱扫描，确定适合NADH和FAD详细扫描图像的两个激发波长和两个检测范围的组合，为这些样品建立了一种适应性测量程序。使用2P-FLIM研究了九只小鼠（三只健康、三只患有慢性败血症、三只患有急性败血症）肾皮质薄片中的选定位置。此外，使用双光子激发荧光（2PEF）强度获得了概览图像。本研究表明，健康的肾切片在荧光寿命特征方面与患有急性败血症的肾切片有很大差异。后者显示内皮质和外皮质之间存在代谢差异，表明在患有急性败血症的小鼠肾脏中，外皮质肾小管细胞将其代谢从氧化磷酸化转变为糖酵解，并在后期恢复，这与患有慢性感染的小鼠情况相同。这些发现表明，2P-FLIM可作为早期败血症诊断和治疗期间监测代谢恢复的有力工具。
