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Transmission Electron Microscopy Analysis on Microbial Ultrathin Sections Prepared by the Ultra-Low Lead Staining Technique

Part of: Micrographia Collection

Published online by Cambridge University Press:  23 July 2021

Zhongquan Jiang ,
Danqing Sun ,
Huize Guan ,
Yutong Sun ,
Menglei Ye ,
Lin Zhang ,
Tingting Gu ,
Jiani Chen ,
Shujie Wang  and
Chunhua Zhang
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Zhongquan Jiang
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
Danqing Sun
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
Huize Guan
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
Yutong Sun
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
Menglei Ye
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
Lin Zhang
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
Tingting Gu
Affiliation:
State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
Jiani Chen
Affiliation:
State Key Laboratory for Mineral Deposits Research, Nanjing University, Nanjing 210046, China
Shujie Wang
Affiliation:
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
Chunhua Zhang
Affiliation:
Laboratory Centre of Life Science, College of Life Science, Nanjing Agricultural University, Nanjing 210095, China
Shimei Wang
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China Jiangsu Provincial Key Lab for Organic Solid Waste Utilization, Nanjing Agricultural University, Nanjing 210095, China
Zhenlei Zhou
Affiliation:
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
Ying Ge*
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China
Zhen Li*
Affiliation:
College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China Jiangsu Provincial Key Lab for Organic Solid Waste Utilization, Nanjing Agricultural University, Nanjing 210095, China
*
*Corresponding authors: Zhen Li, E-mail: lizhen@njau.edu.cn; Ying Ge, E-mail: yingge711@njau.edu.cn
*Corresponding authors: Zhen Li, E-mail: lizhen@njau.edu.cn; Ying Ge, E-mail: yingge711@njau.edu.cn
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Abstract

The staining procedure is critical for investigating intra- and extra-cellular ultrastructure of microorganisms by transmission electron microscopy (TEM). Here, we propose a new ultra-low lead staining (ULLS) technique for preparing the ultrathin sections for TEM analysis. Sections of Enterobacter sp. (bacteria), Aspergillus niger (filamentous fungi), Rhodotorula mucilaginosa (fungi), and Chlamydomonas reinhardtii (microalgae) were tested. Compared with the sections prepared by the typical double-staining technique, ULLS-based sections showed evident advantages: (i) the staining process only required the addition of Pb(NO3)2; (ii) the Pb level during incubation was set as low as 1 mg/L, which had negligible toxicity to most microbial cells; (iii) the Pb cations were added during microbial culture, which avoided complicated sample preparation as in typical double staining. Taking C. reinhardtii as an example, the ULLS technique allowed fine investigation of microbial ultrastructure, e.g., starch granule, mitochondrion, Golgi apparatus, vacuole, and vesicle. Meanwhile, the physiological processes of the cells such as cell lysis and exocytosis were successfully captured, with relatively high contrast. This study hence shows a bright future on preparation of the high-quality ultrathin sections of microbial cells by the ULLS technique.

Keywords

microorganismstransmission electron microscopyultra-low lead stainingultrastructure
Type
Micrographia
Information
Microscopy and Microanalysis , Volume 27 , Issue 5 , October 2021 , pp. 1265 - 1272
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Copyright
Copyright © The Author(s), 2021. Published by Cambridge University Press on behalf of the Microscopy Society of America

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Footnotes

†

These authors contributed equally to this work.

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