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Orson L. Moritz, Beatrice M. Tam, Barry E. Knox, David S. Papermaster; Fluorescent Photoreceptors of Transgenic Xenopus laevis Imaged In Vivo by Two Microscopy Techniques. Invest. Ophthalmol. Vis. Sci. 1999;40(13):3276-3280.
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purpose. To develop a method for imaging individual photoreceptors in an intact
transgenic Xenopus eye, thus allowing in vivo observation of
the effects of various transgenes on photoreceptor development,
degeneration, or both.
methods. Albino and pigmented transgenic Xenopus laevis that
express enhanced green fluorescent protein (GFP) in the major
(“red”) rods were generated. The distribution of GFP throughout the
retina and within the rods was evaluated by confocal microscopy of
frozen sections and immunoelectron microscopy. In vivo images of
photoreceptors were obtained using conventional fluorescence
microscopes to image through the lens of the eye or a laser scanning
confocal microscope to image through the hypopigmented iris of albino
results. Confocal and immunoelectron microscopy of tissue sections showed that
GFP was predominantly localized to the inner segments of the major
rods; a smaller amount was in the outer segments. In a number of
animals, not all the major rods expressed GFP. It was possible
to identify these animals by obtaining fluorescence images of the
retinas of intact, living tadpoles with conventional fluorescence
microscopes, using the lens of the tadpole as part of the optical path.
Confocal images of living animals could be used to visualize the
distribution of GFP within the photoreceptors.
conclusions. The ability to observe individual photoreceptors noninvasively allows
in vivo longitudinal microscopic analysis of photoreceptor development
in transgenic Xenopus tadpoles.
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