Laser-Forschungslabor, LIFE Center, University Hospital, LMU Munich, Planegg, Germany.
Department of Urology, University Hospital, LMU Munich, Munich, Germany.
J Biophotonics. 2021 Jul;14(7):e202100024. doi: 10.1002/jbio.202100024. Epub 2021 Apr 8.
Two-photon fluorescence lifetime imaging is a versatile laboratory technique in the field of biophotonics and its importance is also growing in the field of in vivo diagnostics for medical purposes. After years of experience in dermatology, endoscopic implementations of the technique are now posing new technical challenges. To develop, test, and compare instrumental solutions for this purpose suitable reference samples have been devised and tested. These reference samples can serve as reliable NADH- and FAD-mimicking optical phantoms for 2-photon fluorescence lifetime imaging, as they can be prepared relatively easily with reproducible and stable characteristics for this quite relevant diagnostic technique. The reference samples (mixtures of coumarin 1 and coumarin 6 in ethanol with suitable amounts of 4-hydroxy-TEMPO) have been tuned to exhibit spectral and temporal fluorescence characteristics very similar to those of NADH and FAD, the two molecules most frequently utilized to characterize cell metabolism.
双光子荧光寿命成像技术是生物光子学领域中一种多功能的实验室技术，其在医学体内诊断领域的重要性也在不断增加。在皮肤科积累多年经验后，该技术的内镜实施现在带来了新的技术挑战。为此目的开发、测试和比较仪器解决方案时，设计并测试了合适的参考样本。这些参考样本可用作可靠的 NADH 和 FAD 模拟光学幻像体进行双光子荧光寿命成像，因为它们可以相对容易地用具有可重现且稳定特性的方式制备，这对于非常相关的诊断技术非常重要。参考样本（在乙醇中混合香豆素 1 和香豆素 6 并加入适量的 4-羟基-TEMPO）已被调谐，以显示与 NADH 和 FAD 非常相似的光谱和时间荧光特性，这两种分子是最常用于表征细胞代谢的两种分子。
