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A Kinoshita, H Gong, T Amemiya, K Takaya, M Tozu, Y Ohashi, Y Ohashi; Change Of Elements And Organic Substances In Lens Of Normal Wistar Kyoto Rats . Invest. Ophthalmol. Vis. Sci. 2002;43(13):472.
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Purpose: The Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) microscope is designed to analyze elements and organic substances. The purpose of the present study is to examine with a TOF-SIMS microscope the changes of elements and organic substances in the lenses of rats as they age. Methods: We used 4-, 12-, and 18-month-old normal Wister Kyoto rats. The eyeballs were enucleated and placed in liquid nitrogen without any fixation, cut into semithin sections, and covered with O.C.T compound. Serial semithin sections, 5mm, were cut with a cryo-ultramicrotome and air dried. To confirm the outline of the lens, we observed unstained semithin sections on a silicon wafer plate with a metal microscope. Then TOF-SIMS (TRIFID, Physical Electronics, USA) images were analyzed. Positive ion images were examined with a Ga+source at an acceleration voltage of 15 keV. Secondary ion acceleration voltage of 4.5 keV was used. Results: The present study showed Ca, Na, K, and Mg were contained in the equator more than in the nucleus, and Cu, Zn, Fe, vit E, vit C, palmitic acid, oleic acid and stearic acid were contained in the nuclear zone more than in the equator in 4-, 12-, and 18-month-old rats. In 12-month-old rats, vit E, vit C, palmitic acid, oleic acid and stearic acid in the equator and Ca, vit E, vit C, palmitic acid, oleic acid and stearic acid in the nucleus were lower than those in 4-month-old rats. In 18-month-old rats, Na, Mg, K, Ca, Fe, Cu, Zn in the nucleus were lower than those in 4-month-old rats. Conclusion: We found that vitamins and unsaturated fatty acids decrease with age. TOF-SIMS microscopy showed changes in the concentration of elements in the lenses of rats as they age. It is expected that this tool will be useful in the study of cataract formation.
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