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Daniel Wahl, Yifan Jian, Robert J Zawadzki, Marinko V Sarunic; Wavefront sensorless adaptive optics fluorescence imaging in mouse retina. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5305.
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© ARVO (1962-2015); The Authors (2016-present)
Transgenic mice expressing Green Fluorescent Protein (GFP) as a reporter molecular are particularly important for vision research. The ability to image molecular markers has the potential to accelerate vision research by allowing retina function to be observed in vivo. To achieve cellular resolution imaging in the mouse eye, a wavefront sensorless adaptive optics (WSAO) bio-microscope with fluorescence detection was developed and evaluated by imaging various types of transgenic mice expressing GFP.
We developed a lens-based WSAO mouse retinal imaging system that allowed a more compact optical design. An objective lens focused the light through a glass slide placed on the mouse eye to cancel refraction at the cornea and retain the mouse eye moisture to prevent corneal clouding. The numerical aperture of the imaging system was 0.17, providing an estimated focal waist of 0.9 μm. We utilized a segmented deformable mirror from IrisAO Inc. to perform modal control wavefront sensorless correction, using the intensity of the fluorescence signal as the merit function. Wildtype (c57BL/6) mice, and GFP labeled microglia (Cx3cr1-GFP) and ganglion cell (Thy1-GFP) mice were used in this research. The mice were anesthetized and dilated prior to the imaging session. A generous amount of artificial tear gel was applied in order to maintain corneal hydration. The optical power at the mouse eye was ~100μw which is below the ANSI standard.
Representative WSAO bio-microscope images acquired from a mouse with GFP labeled microglia before and after correction are presented in Fig. 1. Optimization was performed on the small field of view (Top), which was then expanded (Bottom). After WSAO optimization the features in the focal plane were sharper and the overall image brightness was increased.
We have demonstrated a lens based WSAO bio-microscope for high resolution fluorescence imaging in the mouse eye. The WSAO technique is an accessible method of improving image quality with increasing numerical aperture.
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