A preparation of bacterial outer membrane with osmium tetroxide and uranyl acetate co-stain enables improved structural determination by transmission electron microscopy

被引：4
作者：
Sheikh, Aadil [1 ]
Zechmann, Bernd [2 ]
Sayes, Christie M. [3 ]
Taube, Joseph H. [1 ]
Greathouse, K. Leigh [1 ,4 ]
机构：
[1] Baylor Univ, Dept Biol, One Bear Pl 97046, Waco, TX 76798 USA
[2] Baylor Univ, Ctr Microscopy & Imaging, One Bear Pl 97046, Waco, TX 76798 USA
[3] Baylor Univ, Dept Environm Sci, One Bear Pl 97046, Waco, TX 76798 USA
[4] Baylor Univ, Nutr Sci, One Bear Pl 97311, Waco, TX 76798 USA
来源：
MICROSCOPY | 2023年 / 72卷 / 06期
关键词：
bacteria; nanoparticles; outer membrane vesicles; uranyl acetate; osmium tetroxide; transmission electron microscopy; EXTRACELLULAR VESICLES; BIOGENESIS; EXOSOMES;
D O I：
10.1093/jmicro/dfad027
中图分类号：
TH742 [显微镜];
学科分类号：
摘要：
Biological nanoparticles, such as bacterial outer membrane vesicles (OMVs), are routinely characterized through transmission electron microscopy (TEM). In this study, we report a novel method to prepare OMVs for TEM imaging. To preserve vesicular shape and structure, we developed a dual fixation protocol involving osmium tetroxide incubation prior to negative staining with uranyl acetate. Combining osmium tetroxide with uranyl acetate resulted in preservation of sub-50 nm vesicles and improved morphological stability, enhancing characterization of lipid-based nanoparticles by TEM.
引用
收藏
页码：515 / 519
页数：5
相关论文
共 33 条
[1]   Small RNAs in Outer Membrane Vesicles and Their Function in Host-Microbe Interactions [J].
Ahmadi Badi, Sara ;
Bruno, Stefania Paola ;
Moshiri, Arfa ;
Tarashi, Samira ;
Siadat, Seyed Davar ;
Masotti, Andrea .
FRONTIERS IN MICROBIOLOGY, 2020, 11
[2]   Cryo transmission electron microscopy of liposomes and related structures [J].
Almgren, M ;
Edwards, K ;
Karlsson, G .
COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS, 2000, 174 (1-2) :3-21
[3]   Ticket to a bubble ride: Cargo sorting into exosomes and extracellular vesicles [J].
Anand, Sushma ;
Samuel, Monisha ;
Kumar, Sharad ;
Mathivanan, Suresh .
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 2019, 1867 (12)
[4]   Comparative study of pathogenic and non-pathogenic Escherichia coli outer membrane vesicles and prediction of host-interactions with TLR signaling pathways [J].
Behrouzi A. ;
Vaziri F. ;
Riazi Rad F. ;
Amanzadeh A. ;
Fateh A. ;
Moshiri A. ;
Khatami S. ;
Siadat S.D. .
BMC Research Notes, 11 (1)
[5]   Chemical analysis of osmium tetroxide staining in adipose tissue using imaging ToF-SIMS [J].
Belazi, Dalila ;
Sole-Domenech, Santiago ;
Johansson, Bjorn ;
Schalling, Martin ;
Sjovall, Peter .
HISTOCHEMISTRY AND CELL BIOLOGY, 2009, 132 (01) :105-115
[6]   Imaging extracellular vesicles: current and emerging methods [J].
Chuo, Steven Ting-Yu ;
Chien, Jasper Che-Yung ;
Lai, Charles Pin-Kuang .
JOURNAL OF BIOMEDICAL SCIENCE, 2018, 25
[7]   Bacteroides thetaiotaomicron-derived outer membrane vesicles promote regulatory dendritic cell responses in health but not in inflammatory bowel disease [J].
Durant, Lydia ;
Stentz, Regis ;
Noble, Alistair ;
Brooks, Johanne ;
Gicheva, Nadezhda ;
Reddi, Durga ;
O'Connor, Matthew J. ;
Hoyles, Lesley ;
McCartney, Anne L. ;
Man, Ripple ;
Pring, E. Tobias ;
Dilke, Stella ;
Hendy, Philip ;
Segal, Jonathan P. ;
Lim, Dennis N. F. ;
Misra, Ravi ;
Hart, Ailsa L. ;
Arebi, Naila ;
Carding, Simon R. ;
Knight, Stella C. .
MICROBIOME, 2020, 8 (01)
[8]   Bacterial Outer Membrane Vesicles from Dextran Sulfate Sodium-Induced Colitis Differentially Regulate Intestinal UDP-Glucuronosyltransferase 1A1 Partially Through Toll-Like Receptor 4/Mitogen-Activated Protein Kinase/Phosphatidylinositol 3-Kinase Pathway [J].
Gao, Xue-Jiao ;
Li, Ting ;
Wei, Bin ;
Yan, Zhi-Xiang ;
Hu, Nan ;
Huang, Yan-Juan ;
Han, Bei-Lei ;
Wai, Tai-Seng ;
Yang, Wei ;
Yan, Ru .
DRUG METABOLISM AND DISPOSITION, 2018, 46 (03) :292-302
[9]   Natural Nanoparticles: A Particular Matter Inspired by Nature [J].
Griffin, Sharoon ;
Masood, Muhammad Irfan ;
Nasim, Muhammad Jawad ;
Sarfraz, Muhammad ;
Ebokaiwe, Azubuike Peter ;
Schaefer, Karl-Herbert ;
Keck, Cornelia M. ;
Jacob, Claus .
ANTIOXIDANTS, 2018, 7 (01)
[10]  
Griffith W.P., 1974, Plat. Met. Rev, V18, P94
1234