Zhou Haiying, Gunsten Sean P, Zhegalova Natalia G, Bloch Sharon, Achilefu Samuel, Christopher Holley J, Schweppe Daniel, Akers Walter, Brody Steven L, Eades William C, Berezin Mikhail Y
Department of Radiology, Washington University School of Medicine, St. Louis, Missouri.
Cytometry A. 2015 May;87(5):419-27. doi: 10.1002/cyto.a.22658. Epub 2015 Mar 23.
In vivo optical imaging with near-infrared (NIR) probes is an established method of diagnostics in preclinical and clinical studies. However, the specificities of these probes are difficult to validate ex vivo due to the lack of NIR flow cytometry. To address this limitation, we modified a flow cytometer to include an additional NIR channel using a 752 nm laser line. The flow cytometry system was tested using NIR microspheres and cell lines labeled with a combination of visible range and NIR fluorescent dyes. The approach was verified in vivo in mice evaluated for immune response in lungs after intratracheal delivery of the NIR contrast agent. Flow cytometry of cells obtained from the lung bronchoalveolar lavage demonstrated that the NIR dye was taken up by pulmonary macrophages as early as 4-h post-injection. This combination of optical imaging with NIR flow cytometry extends the capability of imaging and enables complementation of in vivo imaging with cell-specific studies.
使用近红外（NIR）探针进行的体内光学成像是临床前和临床研究中一种成熟的诊断方法。然而，由于缺乏近红外流式细胞仪，这些探针的特异性难以在体外进行验证。为了解决这一局限性，我们对一台流式细胞仪进行了改造，使其使用752 nm激光线增加了一个近红外通道。使用近红外微球和用可见光范围和近红外荧光染料组合标记的细胞系对该流式细胞术系统进行了测试。在气管内递送近红外造影剂后评估肺部免疫反应的小鼠体内验证了该方法。对从肺支气管肺泡灌洗中获得的细胞进行流式细胞术分析表明，早在注射后4小时，肺部巨噬细胞就摄取了近红外染料。这种光学成像与近红外流式细胞术的结合扩展了成像能力，并使体内成像能够与细胞特异性研究相互补充。
