Imaging of Norway spruce early somatic embryos with the ESEM,Cryo-SEM and laser scanning microscope |
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| Institution: | 1. Environmental Electron Microscopy Group, Institute of Scientific Instruments of the CAS, Brno, Czech Republic;2. Department of Plant Biology, Faculty of Agronomy, Mendel University in Brno, Brno, Czech Republic;1. Department of General and Inorganic Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, Pardubice 532 10, Czech Republic;2. Department of Inorganic Technology, Faculty of Chemical Technology, University of Pardubice, Doubravice 41, Pardubice 532 10, Czech Republic;3. Joint Laboratory of Solid State Chemistry of Institute of Macromolecular Chemistry of the Academy of Sciences of the Czech Republic and University of Pardubice, Faculty of Chemical Technology, Studentská 95, Pardubice 532 10, Czech Republic;1. Department of Life Sciences, Sogang University, Seoul, 04107, Republic of Korea;2. Department of Bioindustry and Bioresource Engineering, Plant Engineering Research Institute, Sejong University, Seoul, 05006, Republic of Korea;1. College of Resources and Environment, Hunan Agricultural University, Changsha, 410128, PR China;2. Hunan Engineering & Technology Research Center for Irrigation Water Purification, Changsha, 410128, PR China;3. College of Environmental Science and Engineering, Hunan University, Changsha, 410082, PR China |
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| Abstract: | This article describes the surface structure of Norway spruce early somatic embryos (ESEs) as a typical culture with asynchronous development. The microstructure of extracellular matrix covering ESEs were observed using the environmental scanning electron microscope as a primary tool and using the scanning electron microscope with cryo attachment and laser electron microscope as a complementary tool allowing our results to be proven independently. The fresh samples were observed in conditions of the air environment of the environmental scanning electron microscope (ESEM) with the pressure from 550 Pa to 690 Pa and the low temperature of the sample from ?18 °C to ?22 °C. The samples were studied using two different types of detector to allow studying either the thin surface structure or material composition. The scanning electron microscope with cryo attachment was used for imaging frozen extracellular matrix microstructure with higher resolution. The combination of both electron microscopy methods was suitable for observation of “native” plant samples, allowing correct evaluation of our results, free of error and artifacts. |
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| Keywords: | ESEM Cryo-SEM Bright field/dark field microscopy Extracellular matrix Somatic embryogenesis |
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