基于辐射传递方程的荧光分子层析成像在果蝇蛹中干细胞的体内监测。
In vivo monitoring of stem cells in Drosophila pupae using the radiative transfer equation-based fluorescence molecular tomography.
机构信息
J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL 32611, USA.
出版信息
Mol Imaging Biol. 2011 Oct;13(5):868-73. doi: 10.1007/s11307-010-0434-6.
PURPOSE
The aim of this study is to realize in vivo monitoring of DSRed-expressing cell distribution in Drosophila pupae using fluorescence molecular tomography (FMT).
PROCEDURES
The radiative transfer equation (RTE)-based FMT reconstruction algorithm is implemented using finite element method for mesoscopic- or millimeter-scale imaging. The RTE algorithm is validated using both simulated and phantom experimental data. For the in vivo experiments, DsRed fluorescent reporter was inserted into the middle of irradiation-responsive enhancer region (IRER) of Drosophila pupae and expressed only in IRER-open cells.
RESULTS
Quantitatively accurate fluorescence images can be reconstructed from both simulated and phantom data. The in vivo images obtained agree well with the in vitro images obtained from confocal microscopy both qualitatively and semi-quantitatively.
CONCLUSION
DsRed-bearing stem cells in Drosophila pupae can be markedly imaged using our FMT approach. This study suggests that the technique described can be used for dynamic in vivo monitoring of biological events in mesoscopic-scale animals and facilitates basic biologic research such as genetics, epigenetics, and stem cells.
目的
本研究旨在利用荧光分子断层扫描(FMT)实现对果蝇蛹中表达 DSRed 的细胞分布的活体监测。
方法
采用有限元法实现基于辐射传输方程(RTE)的 FMT 重建算法,用于介观或毫米尺度成像。RTE 算法通过模拟和体模实验数据进行验证。对于体内实验,将 DsRed 荧光报告基因插入果蝇蛹的辐射响应增强子区域(IRER)的中间,仅在 IRER 开放细胞中表达。
结果
从模拟和体模数据中可以重建出定量准确的荧光图像。从共聚焦显微镜获得的体内图像在定性和半定量方面与体外图像吻合良好。
结论
我们的 FMT 方法可以显著地对果蝇蛹中的 DsRed 标记干细胞进行成像。这项研究表明,所描述的技术可用于在介观尺度动物中进行动态活体监测生物事件,并促进遗传学、表观遗传学和干细胞等基础生物学研究。