使用光纤共焦荧光显微镜实时评估超声介导的药物输送。
Real-time assessment of ultrasound-mediated drug delivery using fibered confocal fluorescence microscopy.
机构信息
Laboratory for Molecular and Functional Imaging: From Physiology to Therapy, FRE 3313-CNRS and University Bordeaux Segalen, 146, rue Léo Saignat, Case 117, 33076, Bordeaux cedex, France.
出版信息
Mol Imaging Biol. 2013 Feb;15(1):3-11. doi: 10.1007/s11307-012-0568-9.
PURPOSE
Transport across the plasma membrane is a critical step of drug delivery for weakly permeable compounds with intracellular mode of action. The purpose of this study is to demonstrate real-time monitoring of ultrasound (US)-mediated cell-impermeable model drug uptake with fibered confocal fluorescence microscopy (FCFM).
PROCEDURES
An in vitro setup was designed to combine a mono-element US transducer, a cell chamber with a monolayer of tumor cells together with SonoVue microbubbles, and a FCFM system. The cell-impermeable intercalating dye, SYTOX Green, was used to monitor US-mediated uptake.
RESULTS
The majority of the cell population showed fluorescence signal enhancement 10 s after US onset. The mean rate constant k of signal enhancement was calculated to be 0.23 ± 0.04 min(-1).
CONCLUSIONS
Feasibility of real-time monitoring of US-mediated intracellular delivery by FCFM has been demonstrated. The method allowed quantitative assessment of model drug uptake, holding great promise for further local drug delivery studies.
目的
对于具有细胞内作用模式的弱渗透性化合物，跨质膜的转运是药物递送的关键步骤。本研究旨在通过纤维共聚焦荧光显微镜（FCFM）实时监测超声（US）介导的细胞不可渗透的模型药物摄取。
方法
设计了一种体外装置，将单元件超声换能器、单层肿瘤细胞的细胞室与 SonoVue 微泡以及 FCFM 系统结合在一起。使用不透细胞的嵌入染料 SYTOX Green 来监测 US 介导的摄取。
结果
大多数细胞群在 US 开始后 10 秒显示荧光信号增强。计算出的信号增强的平均速率常数 k 为 0.23±0.04 min（-1）。
结论
通过 FCFM 实时监测 US 介导的细胞内递药的可行性已得到证明。该方法允许对模型药物摄取进行定量评估，为进一步的局部药物递送研究提供了很大的前景。
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