Application News No.P100 Electron Probe Microanalyzer Analysis of Biological Tissues Using the EPMA-8050G LAAN-A-EP-E007 n Introduction n Cross Sections of Biological Tissues Fig. 1 Cross Sections of : SHIMADZU (Shimadzu Corporation)

Application News No.P100 Electron Probe Microanalyzer Analysis of Biological Tissues Using the EPMA-8050G LAAN-A-EP-E007 n Introduction n Cross Sections of Biological Tissues Fig. 1 Cross Sections of Pathological Lung Tissues Biological tissues consist primarily of soft tissues and hard tissues. Bones, teeth and other hard tissues are formed primarily of calcium carbonate. In contrast, soft tissues are formed mainly of proteins. Metallic elements existing in biological soft tissues are strongly related to the growth (development) of these tissues. Understanding the concentrations and distributions of metallic elements plays a role in assessing developmental environments and states of health. Human living and working environments contain naturally derived fine particles, as well as fine particles originating from work sites. Exposure to these particles sometimes brings the risk of impaired human health. Here, we introduce an example of the analysis of biological tissues using the EPMA-8050G (FE-EPMA). Biological soft tissues are prone to destruction, deformation and other damage from the heat resulting from electron beam exposure. However, by using a special carbon stand (vitreous carbon with an average coarseness of approximately 3 nm), a biological tissue cross section approximately 2 µm thick can be closely afﬁxed onto it without using adhesives. In this case, the heat from the electron beam penetrating the biological tissue cross section is absorbed by the carbon stand, so the damage to the samples is signiﬁcantly reduced. Fig. 1 shows the results of repeated analysis with the EPMA of cross sections of pathological tissue from the lungs of a person with a history of exposure to dust. There is virtually no damage to the biological tissue cross sections from exposure to the electron beam, so the same area can be analyzed repeatedly. OM Image 480×360 μm 500 μm COMPO 25.0 kV 2.0×1.5 mm 500 μm N Ka 25.0kV 2.0×1.5 mm 11400 (counts) 1800 10200 9000 7800 6600 5400 4200 3000 50 μm COMPO 25.0 kV 300×225 μm 50 μm Mg Ka 25.0 kV 300×225 μm 820 (counts) 340 760 700 640 580 520 460 400 50 μm Al Ka 25.0 kV 300×225 μm 3000 (counts) 600 2700 2400 2100 1800 1500 1200 900 50 μm Si Ka 25.0 kV 300×225 μm 1700 (counts) 100 1500 1300 1100 900 700 500 300 50 μm Ti Ka 25.0 kV 300×225 μm 1240 (counts) 440 1140 1040 940 840 740 640 540 50 μm Fe Ka 25.0 kV 300×225 μm 600 (counts) 200 550 500 450 400 350 300 250

Application News No. For Research Use Only. Not for use in diagnostic procedures. The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. The information contained herein is provided to you "as is" without warranty of any kind including without limitation warranties as to its accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. This publication is based upon the information available to Shimadzu on or before the date of publication, and subject to change without notice. © Shimadzu Corporation, 2015 www.shimadzu.com/an/ P100 First Edition: Apr. 2015 n Toxicological Evaluation of Nanoparticles When people breathe the air, they inhale fine particles of various sizes. Of the fine particles inhaled, those ranging in size from approximately 0.1 µm to 1 µm are not easily trapped by mucus, and are said to reach the alveoli of the lungs easily. These fine particles appear as if they are aggregates approximately 10 µm in size wrapped in thin veils. The reason is said to be that rather than being captured and existing in the alveolar walls, they are subject to phagocytosis by the alveolar macrophages, and toxic compounds are then released to the surroundings during macrophage necrosis. In Fig. 2, an intracellular distribution of fine particles (Mg, Al, Si, Ti, and Fe) ranging in size from approximately 0.1 µm to 1 µm can be conﬁrmed. In Fig. 3, it is also possible to conﬁrm the existence of ultraﬁne particles (Fe) approximately 50 nm in size. The distribution of speciﬁc elements in the 1 µm range is expected to be of use in pathological clarification of toxicity related to the accumulation of ﬁne particles (nanoparticles) of metallic elements. Fig. 3 Elemental Mapping of Nanoparticles Fig. 2 Alveolar Macrophages 2 μm COMPO 25.0 kV ×10000 2 μm Mg Ka 25.0 kV ×10000 460 (counts) 140 420 380 340 300 260 220 180 2 μm Al Ka 25.0 kV ×10000 11500 (counts) 300 10100 8700 7300 5900 4500 3100 1700 2 μm Si Ka 25.0 kV ×10000 4850 (counts) 50 4250 3650 3050 2450 1850 1250 650 2 μm Ti Ka 25.0 kV ×10000 1020 (counts) 220 920 820 720 620 520 420 320 2 μm Fe Ka 25.0 kV ×10000 740 (counts) 100 660 580 500 420 340 260 180 2 μm COMPO 25.0 kV ×10000 500 nm COMPO 25.0kV ×60000 500 nm SE 25.0 kV ×60000 500 nm Fe Ka 25.0 kV ×60000 403 (counts) 10 220 190 160 130 100 70 40 500 nm Si Ka 25.0 kV ×60000 181 (counts) 20 132 116 100 84 68 52 36 500 nm Al Ka 25.0 kV ×60000 364 (counts) 40 292 256 220 184 148 112 76 Samples Provided by: Center for Instrumental Analysis, Niigata University