Noninvasive high-resolution in vivo imaging of cell biology in the anterior chamber of the mouse eye

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Abstract

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.

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Figure 1: Imaging setup for noninvasive in vivo imaging in the anterior chamber of the eye.
Figure 2: Noninvasive in vivo imaging in the anterior chamber of the eye.
Figure 3: Custom-built stabilizer of the mouse eye for noninvasive in vivo imaging.
Figure 4: Perfusion of the anterior chamber of the mouse eye.
Figure 5: Morphological characterization of a pancreatic islet graft by imaging reflection and GFP.
Figure 6: Imaging of tissue vascularization.
Figure 7: Imaging of cell death.
Figure 8: Loading of cells within the anterior chamber of the eye with calcium indicators.

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Acknowledgements

This study was supported by grants DK-58508 and DK-075487 (to A.C.) from the US National Institutes of Health, Juvenile Diabetes Research Foundation International grant 3-2007-73 (to S.S.) and 4-2004-361, the Swedish Research Council, the Novo Nordisk Foundation, Karolinska Institutet, the Swedish Diabetes Association, The Family Knut and Alice Wallberg Foundation, Eurodia (FP6-518153), European Foundation for the Study of Diabetes, the EFSD/Lilly Research Program, Berth von Kantzow's Foundation, the Family Erling-Persson Foundation and the Diabetes Research Institute Foundation (Hollywood, FL).

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The first two authors contributed equally to this work.

Correspondence to Stephan Speier.

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Speier, S., Nyqvist, D., Köhler, M. et al. Noninvasive high-resolution in vivo imaging of cell biology in the anterior chamber of the mouse eye. Nat Protoc 3, 1278–1286 (2008) doi:10.1038/nprot.2008.118

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