U Schlötzer-Schrehardt1, K H Körtje, C Erb. 1. Department of Ophthalmology, University of Erlangen-Nürnberg, Erlangen, Germany. ursula.schloetzer@augen.imed.uni-erlangen.de
Abstract
PURPOSE: To obtain more information on the basic nature of the pathological matrix product accumulating in pseudoexfoliation (PEX) syndrome by analyzing its elemental composition at the subcellular level. METHODS: Energy-filtering transmission electron microscopy (EFTEM), combining the two microanalytical techniques of electron spectroscopic imaging (ESI) and energy-loss spectroscopy (EELS), were performed on ultrathin sections of lens specimens with PEX syndrome using a transmission electron microscope equipped with an integrated electron energy filter. EFTEM is based on inner shell ionization of elements present in the sample giving rise to characteristic signals in well-defined energy-loss regions. The EEL-spectra, demonstrating the presence of a particular element by its specific electron energy-loss edge, were recorded with an integrated scintillator-photomultiplier-system. ESI generated graphic images of elemental localization in the sections after a process of background correction with an IBAS image analysis program. Energy-dispersive X-ray (EDX) analysis of PEX deposits on hydrated lenses was conducted by variable pressure scanning electron microscopy. RESULTS: The ESI element distribution images of both intracapsular and supracapsular PEX material displayed high signals for nitrogen, sulfur, calcium, chlorine, and zinc in clear association with the PEX fibrils. The corresponding EEL-spectra confirmed the data obtained by ESI and showed the presence of the element-specific energy-loss edges. The presence of these elements in PEX fibrils was further confirmed by EDX analysis. No specific signals were obtained for phosphorus, oxygen, or aluminum. CONCLUSIONS: This study demonstrates the presence of nitrogen, sulfur, chlorine, zinc, and calcium both in mature and in aggregating PEX fibrils of the lens capsule. EFTEM proved to be a highly sensitive method for the microanalytical study of biological material with unknown composition, such as PEX material, at the subcellular level.
PURPOSE: To obtain more information on the basic nature of the pathological matrix product accumulating in pseudoexfoliation (PEX) syndrome by analyzing its elemental composition at the subcellular level. METHODS: Energy-filtering transmission electron microscopy (EFTEM), combining the two microanalytical techniques of electron spectroscopic imaging (ESI) and energy-loss spectroscopy (EELS), were performed on ultrathin sections of lens specimens with PEX syndrome using a transmission electron microscope equipped with an integrated electron energy filter. EFTEM is based on inner shell ionization of elements present in the sample giving rise to characteristic signals in well-defined energy-loss regions. The EEL-spectra, demonstrating the presence of a particular element by its specific electron energy-loss edge, were recorded with an integrated scintillator-photomultiplier-system. ESI generated graphic images of elemental localization in the sections after a process of background correction with an IBAS image analysis program. Energy-dispersive X-ray (EDX) analysis of PEX deposits on hydrated lenses was conducted by variable pressure scanning electron microscopy. RESULTS: The ESI element distribution images of both intracapsular and supracapsular PEX material displayed high signals for nitrogen, sulfur, calcium, chlorine, and zinc in clear association with the PEX fibrils. The corresponding EEL-spectra confirmed the data obtained by ESI and showed the presence of the element-specific energy-loss edges. The presence of these elements in PEX fibrils was further confirmed by EDX analysis. No specific signals were obtained for phosphorus, oxygen, or aluminum. CONCLUSIONS: This study demonstrates the presence of nitrogen, sulfur, chlorine, zinc, and calcium both in mature and in aggregating PEX fibrils of the lens capsule. EFTEM proved to be a highly sensitive method for the microanalytical study of biological material with unknown composition, such as PEX material, at the subcellular level.
Authors: Tin Aung; Mineo Ozaki; Takanori Mizoguchi; R Rand Allingham; Zheng Li; Aravind Haripriya; Satoko Nakano; Steffen Uebe; Jeffrey M Harder; Anita S Y Chan; Mei Chin Lee; Kathryn P Burdon; Yury S Astakhov; Khaled K Abu-Amero; Juan C Zenteno; Yildirim Nilgün; Tomasz Zarnowski; Mohammad Pakravan; Leen Abu Safieh; Liyun Jia; Ya Xing Wang; Susan Williams; Daniela Paoli; Patricio G Schlottmann; Lulin Huang; Kar Seng Sim; Jia Nee Foo; Masakazu Nakano; Yoko Ikeda; Rajesh S Kumar; Morio Ueno; Shin-ichi Manabe; Ken Hayashi; Shigeyasu Kazama; Ryuichi Ideta; Yosai Mori; Kazunori Miyata; Kazuhisa Sugiyama; Tomomi Higashide; Etsuo Chihara; Kenji Inoue; Satoshi Ishiko; Akitoshi Yoshida; Masahide Yanagi; Yoshiaki Kiuchi; Makoto Aihara; Tsutomu Ohashi; Toshiya Sakurai; Takako Sugimoto; Hideki Chuman; Fumihiko Matsuda; Kenji Yamashiro; Norimoto Gotoh; Masahiro Miyake; Sergei Y Astakhov; Essam A Osman; Saleh A Al-Obeidan; Ohoud Owaidhah; Leyla Al-Jasim; Sami Al Shahwan; Rhys A Fogarty; Paul Leo; Yaz Yetkin; Çilingir Oğuz; Mozhgan Rezaei Kanavi; Afsaneh Nederi Beni; Shahin Yazdani; Evgeny L Akopov; Kai-Yee Toh; Gareth R Howell; Andrew C Orr; Yufen Goh; Wee Yang Meah; Su Qin Peh; Ewa Kosior-Jarecka; Urszula Lukasik; Mandy Krumbiegel; Eranga N Vithana; Tien Yin Wong; Yutao Liu; Allison E Ashley Koch; Pratap Challa; Robyn M Rautenbach; David A Mackey; Alex W Hewitt; Paul Mitchell; Jie Jin Wang; Ari Ziskind; Trevor Carmichael; Rangappa Ramakrishnan; Kalpana Narendran; Rangaraj Venkatesh; Saravanan Vijayan; Peiquan Zhao; Xueyi Chen; Dalia Guadarrama-Vallejo; Ching Yu Cheng; Shamira A Perera; Rahat Husain; Su-Ling Ho; Ulrich-Christoph Welge-Luessen; Christian Mardin; Ursula Schloetzer-Schrehardt; Axel M Hillmer; Stefan Herms; Susanne Moebus; Markus M Nöthen; Nicole Weisschuh; Rohit Shetty; Arkasubhra Ghosh; Yik Ying Teo; Matthew A Brown; Ignacio Lischinsky; Jonathan G Crowston; Michael Coote; Bowen Zhao; Jinghong Sang; Nihong Zhang; Qisheng You; Vera Vysochinskaya; Panayiota Founti; Anthoula Chatzikyriakidou; Alexandros Lambropoulos; Eleftherios Anastasopoulos; Anne L Coleman; M Roy Wilson; Douglas J Rhee; Jae Hee Kang; Inna May-Bolchakova; Steffen Heegaard; Kazuhiko Mori; Wallace L M Alward; Jost B Jonas; Liang Xu; Jeffrey M Liebmann; Balram Chowbay; Elke Schaeffeler; Matthias Schwab; Fabian Lerner; Ningli Wang; Zhenglin Yang; Paolo Frezzotti; Shigeru Kinoshita; John H Fingert; Masaru Inatani; Kei Tashiro; André Reis; Deepak P Edward; Louis R Pasquale; Toshiaki Kubota; Janey L Wiggs; Francesca Pasutto; Fotis Topouzis; Michael Dubina; Jamie E Craig; Nagahisa Yoshimura; Periasamy Sundaresan; Simon W M John; Robert Ritch; Michael A Hauser; Chiea-Chuen Khor Journal: Nat Genet Date: 2015-02-23 Impact factor: 38.330
Authors: Josef A Schroeder; Christian Weingart; Brigitte Coras; Ingrid Hausser; Stephan Reinhold; Matthias Mack; Volker Seybold; Thomas Vogt; Bernhard Banas; Ferdinand Hofstaedter; Bernhard K Krämer Journal: Clin J Am Soc Nephrol Date: 2008-04-02 Impact factor: 8.237