小鼠眼前房中细胞生物学的无创高分辨率体内成像。
Noninvasive high-resolution in vivo imaging of cell biology in the anterior chamber of the mouse eye.
作者信息
Speier Stephan, Nyqvist Daniel, Köhler Martin, Caicedo Alejandro, Leibiger Ingo B, Berggren Per-Olof
机构信息
The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital L1, SE-17176 Stockholm, Sweden.
出版信息
Nat Protoc. 2008;3(8):1278-86. doi: 10.1038/nprot.2008.118.
There is clearly a demand for an experimental platform that enables cell biology to be studied in intact vascularized and innervated tissue in vivo. This platform should allow observations of cells noninvasively and longitudinally at single-cell resolution. For this purpose, we use the anterior chamber of the mouse eye in combination with laser scanning microscopy (LSM). Tissue transplanted to the anterior chamber of the eye is rapidly vascularized, innervated and regains function. After transplantation, LSM through the cornea allows repetitive and noninvasive in vivo imaging at cellular resolution. Morphology, vascularization, cell function and cell survival are monitored longitudinally using fluorescent proteins and dyes. We have used this system to study pancreatic islets, but the platform can easily be adapted for studying a variety of tissues and additional biological parameters. Transplantation to the anterior chamber of the eye takes 25 min, and in vivo imaging 1-5 h, depending on the features monitored.
显然需要一个实验平台，以便能够在完整的、具有血管化和神经支配的体内组织中研究细胞生物学。该平台应允许以单细胞分辨率对细胞进行非侵入性的纵向观察。为此，我们将小鼠眼睛的前房与激光扫描显微镜（LSM）结合使用。移植到眼睛前房的组织会迅速血管化、神经化并恢复功能。移植后，通过角膜的激光扫描显微镜可在细胞分辨率下进行重复的非侵入性体内成像。使用荧光蛋白和染料对形态、血管化、细胞功能和细胞存活进行纵向监测。我们已使用该系统研究胰岛，但该平台可轻松适用于研究各种组织和其他生物学参数。移植到眼睛前房需要25分钟，体内成像需要1 - 5小时，具体取决于所监测的特征。
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