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. 2021 Jan 8;11:228. doi: 10.1038/s41598-020-80546-0

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Experimental setup of SE-ADM system and observation of cultured mammalian cells with PM2.5. (a) A schematic diagram of the SE-ADM system based on high-resolution FE-SEM. The liquid-sample holder with cultured cells with PM2.5 added was mounted on the pre-amplifier-attached stage, which was introduced into the SEM specimen chamber. The scanning EB was applied to the W-coated SiN film at a low acceleration voltage. The measurement terminal under the holder detected the electrical signal through the liquid specimens. (b) Overview of the liquid-sample holder with cultured mammalian cells with PM2.5 aggregates. OBA9 or 4T1E/M3 cells were on the upper SiN film and the W-coated side was irradiated with the scanning EB. (c) A conceptual diagram of the PM2.5 aggregates in a cell. (d) A low magnification SE-ADM image of untreated OBA9 cells in medium (1000×) with a 7-kV EB and − 9 V bias. Nuclei indicated by red arrows were observed in this image. (e) A high magnification image of OBA9 cells (5000×). Intracellular membranes and vesicles were observed. (f) An SE-ADM image of OBA9 cells with added PM2.5 at an electron beam acceleration of 7 kV (2000×). Five hours after the addition of PM2.5, the black aggregates were detected in the whole visual field, indicating aggregated PM2.5 in the cells. (g) A high-magnification image (10,000×) of a PM2.5 aggregate in the red square in (f). Scale bars, 10 μm in (d, f) and 1 μm in (e, g).