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Nanoparticle suspensions enclosed in methylcellulose : a new approach for quantifying nanoparticles in transmission electron microscopy

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Date
04/05/2016
Author
Hacker, Christian
Asadi, Jalal
Pliotas, Christos
Ferguson, Sophie Grace Alicia
Sherry, Lee
Marius, Phedra
Tello, Javier
Jackson, David
Naismith, James Henderson
Lucocq, John Milton
Keywords
Electron microscopy
Liposome
Methylcellulose
Nanoparticles
Negative stain
Quantification
Stereology
Virus
QD Chemistry
QH301 Biology
RC Internal medicine
NDAS
BDC
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Abstract
Nanoparticles are of increasing importance in biomedicine but quantification is problematic because current methods depend on indirect measurements at low resolution. Here we describe a new high-resolution method for measuring and quantifying nanoparticles in suspension. It involves premixing nanoparticles in a hydrophilic support medium (methylcellulose) before introducing heavy metal stains for visualization in small air-dried droplets by transmission electron microscopy (TEM). The use of methylcellulose avoids artifacts of conventional negative stain-TEM by (1) restricting interactions between the nanoparticles, (2) inhibiting binding to the specimen support films and (3) reducing compression after drying. Methylcellulose embedment provides effective electron imaging of liposomes, nanodiscs and viruses as well as comprehensive visualization of nanoparticle populations in droplets of known size. These qualities facilitate unbiased sampling, rapid size measurement and estimation of nanoparticle numbers by means of ratio counting using a colloidal gold calibrant. Specimen preparation and quantification take minutes and require a few microliters of sample using only basic laboratory equipment and a standard TEM.
Citation
Hacker , C , Asadi , J , Pliotas , C , Ferguson , S G A , Sherry , L , Marius , P , Tello , J , Jackson , D , Naismith , J H & Lucocq , J M 2016 , ' Nanoparticle suspensions enclosed in methylcellulose : a new approach for quantifying nanoparticles in transmission electron microscopy ' , Scientific Reports , vol. 6 , 25275 . https://doi.org/10.1038/srep25275
Publication
Scientific Reports
Status
Peer reviewed
DOI
https://doi.org/10.1038/srep25275
ISSN
2045-2322
Type
Journal article
Rights
This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
Description
This work was supported by the University of St Andrews and Nanomorphomics group funds. The work forms part of an International Patent Application No. PCT/GB2015/052482.
Collections
  • University of St Andrews Research
URI
http://hdl.handle.net/10023/8790

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