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用硅涂层上转换纳米粒子成像间隙连接。

Imaging gap junctions with silica-coated upconversion nanoparticles.

机构信息

Division of Bioengineering, Faculty of Engineering, National University of Singapore, Singapore.

出版信息

Med Biol Eng Comput. 2010 Oct;48(10):1033-41. doi: 10.1007/s11517-010-0618-x. Epub 2010 May 21.

Abstract

Upconversion nanoparticles (UCN) that are excited in the near infrared (NIR) region were synthesized and modified to enable their application to biological systems for imaging. The UCN obtained are oleic acid capped and hence hydrophobic in nature. Since the particles were to be used for imaging cells, a surface modification to make them hydrophilic and biocompatible was performed. Silica coating was chosen for the modification due to the possibility to further functionalize the surface and conjugate biomolecules. Cardiac cells which are capable of forming gap junctions were selected to be labeled. Gap junction specific antibodies were conjugated to the silica-coated UCN. The fluorescence emission spectrum of the particles was obtained with a continuous wave 980 nm laser and size of the particles before and after coating was determined to be 30 and 50 nm, respectively, by TEM. A covalent coupling method was used to bind the gap junction specific antibody to the nanoparticles. The fluorescence imaging experiments were carried out on cardiomyoblast cells and co-culture of bone marrow stem cells/cardiomyoblast cells after incubation with the antibody-modified UCN. Images of the particles after incubation with cardiac cells obtained over days demonstrated the potentials of the UCN for fluorescence imaging.

摘要

上转换纳米粒子(UCN)在近红外(NIR)区域被激发,被合成并修饰以使其能够应用于生物系统的成像。所获得的 UCN 被油酸帽封,因此具有疏水性。由于这些颗粒将用于细胞成像,因此进行了表面修饰以使其亲水和生物相容。由于有可能进一步对表面进行功能化和连接生物分子,因此选择了硅涂层进行修饰。选择能够形成缝隙连接的心脏细胞进行标记。将缝隙连接特异性抗体连接到硅涂层的 UCN 上。通过连续波 980nm 激光获得颗粒的荧光发射光谱,并通过 TEM 分别确定颗粒在涂层前后的尺寸为 30nm 和 50nm。使用共价偶联方法将缝隙连接特异性抗体结合到纳米颗粒上。在用抗体修饰的 UCN 孵育后,对心肌细胞和骨髓干细胞/心肌细胞共培养物进行荧光成像实验。用荧光显微镜在孵育数天后对与心脏细胞孵育后的颗粒进行成像,证明了 UCN 用于荧光成像的潜力。

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