In vivo fluorescent imaging of the mouse retina using adaptive optics - PubMed (original) (raw)

In vivo fluorescent imaging of the mouse retina using adaptive optics

David P Biss et al. Opt Lett. 2007.

Abstract

In vivo imaging of the mouse retina using visible and near infrared wavelengths does not achieve diffraction-limited resolution due to wavefront aberrations induced by the eye. Considering the pupil size and axial dimension of the eye, it is expected that unaberrated imaging of the retina would have a transverse resolution of 2 microm. Higher-order aberrations in retinal imaging of human can be compensated for by using adaptive optics. We demonstrate an adaptive optics system for in vivo imaging of fluorescent structures in the retina of a mouse, using a microelectromechanical system membrane mirror and a Shack-Hartmann wavefront sensor that detects fluorescent wavefront.

PubMed Disclaimer

Figures

Fig. 1

Fig. 1

(Color online) Experimental setup. DM, deformable mirror, SHWS, Shack–Hartmann wavefront sensor. PMT, photomultiplier tube for confocal detection of fluorescence, HS, polygon scanner for the horizontal scan. VS, galvanometric mirror for the vertical scan, OBJ, imaging objective, EYE, mouse eye sample.

Fig. 2

Fig. 2

In vivo images of retinal blood vessels injected with an Evan’s Blue solution and measured SHWS spot displacements (arrows) with calculated wavefront (gray-scale image). The pupil at the SHWS is 4 mm across, and lenslets are 300 _μ_m in diameter and have a 7 mm focal length. (a) Image without AO correction. (b) Wavefront measurements before AO correction. (c) Image with AO correction. (d) Wavefront measurements after AO correction. The wavefront scale bar represents −3 to 3 _μ_m.

Fig. 3

Fig. 3

In vivo images of GFP-labeled microglia in the retina. (a) Without AO correction. A line plot is given for the region of the image that corresponds to the white line. (b) With AO correction. In (a) and (b) pixel intensity from the original, unprocessed image along the white line is plotted on the same scale. (c) Ex vivo flat-mount image of the same retina. The dimension bar is 50 _μ_m. Both inlayed line plots have the same x and y scale.

Similar articles

Cited by

References

    1. Paques M, Simonutti M, Roux MJ, Picaud S, Levavassuer E, Bellman C, Sahel JA. Vision Res. 2006;46:1336. - PubMed
    1. Higashide T, Kawaguchi I, Ohkubo S, Takeda H, Sugiyama K. Invest Opthalmol Visual Sci. 2006;47:2943. - PubMed
    1. García de la Cera E, Rodríguez G, Llorente L, Schaeffel F, Marcos S. Vision Res. 2006;46:2546. - PubMed
    1. Liang J, Williams DR, Miller D. J Opt Soc Am A. 1997;14:2884. - PubMed
    1. Roorda A, Williams DR. Nature. 1999;397:520. - PubMed

Publication types

MeSH terms

Grants and funding

LinkOut - more resources