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Longitudinal Profile of Retinal Ganglion Cell Damage after Optic Nerve Crush with Blue-Light Confocal Scanning Laser Ophthalmoscopy

¹From the Hamilton Glaucoma Center, Department of Ophthalmology, University of California, San Diego, California; the ²Department of Ophthalmology and Visual Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong; and the ³Department of Ophthalmology, Center of Eye Research, Australia, University of Melbourne, Melbourne, Australia.

PURPOSE. To investigate the long-term longitudinal profile of retinal ganglion cell (RGC) damage after optic nerve crush with a new technique for in vivo imaging of RGCs.

METHODS. A blue-light confocal scanning laser ophthalmoscope (bCSLO; 460 nm excitation, 490 nm detection) was used to image Thy-1 CFP mice aged 6 to 9 months (n = 5) before optic nerve crush, weekly after crush for 3 weeks, and at weeks 10 and 50 after optic nerve crush. A sham procedure was performed in the contralateral eye, and it was imaged as a control. Corresponding retinal areas before and after optic nerve crush were compared, and the fluorescent spots were counted manually. The longitudinal profile of RGC degeneration was modeled and compared with one-phase and two-phase exponential decay equations.

RESULTS. A significant and progressive loss of fluorescent spots was found after optic nerve crush with 18.6% ± 2.3%, 11.3% ± 3.4%, 8.8% ± 5.3%, 4.2% ± 3.1%, and 3.3% ± 2.1% of Thy-1–expressing RGCs remaining at weeks 1, 2, 3, 10, and 50, respectively, after optic nerve crush (P < 0.001; n = 5). There was no change in the fluorescence density in the contralateral control (P = 0.893). Two-phase exponential decay (y = 0.03 + 0.83e^–2.78t + 0.14e^–0.30t) was a better fit than one-phase exponential decay (y = 0.94e^–1.93t + 0.06; P = 0.003) equations, with half-lives of fast phase and slow phase of 1.7 days and 16.3 days, respectively.

CONCLUSIONS. The longitudinal profile of RGC degeneration after optic nerve crush is characterized by a two-phase exponential decay model. bCSLO imaging provides an efficient and noninvasive approach to the longitudinal study of progressive RGC damage.

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				C K S Leung, J D Lindsey, L Chen, Q Liu, and R N Weinreb Longitudinal profile of retinal ganglion cell damage assessed with blue-light confocal scanning laser ophthalmoscopy after ischaemic reperfusion injury Br J Ophthalmol, July 1, 2009; 93(7): 964 - 968. [Abstract] [Full Text] [PDF]