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NIST - NCL Joint Assay Protocol, PCC-7

By John Bonevich1, Wolfgang Haller1

Leidos Biomedical Research, Inc.

Measuring the Size of Nanoparticles Using Transmission Electron Microscopy (TEM)

Listed in Datasets | publication by group NCL Protocols

Version 1.0 - published on 04 May 2020 doi:10.17917/C7N7-N938 - cite this

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Description

Transmission electron microscopy (TEM) has been found to be an excellent tool for characterizing the size of nanoparticles.  This assay protocol outlines procedures for sample preparation and the determination of mean nanoparticle size (in projection) using TEM.  Although the projected particle size is the primary determinant of the measured particle diameter, other parameters can impact these measurements and influence the measured size.  Therefore, guidelines for making successful size measurements in the nanometer-size range are provided, as well as a discussion of relevant standards and data analysis.  This protocol can be applied to any suitable TEM instrument with image recording capability.  

The specimen preparation protocol has been specifically tailored for application to NIST Reference Materials RM8011, RM8012 and RM8013, which consist of negatively charged citrate-stabilized gold nanoparticles in aqueous solution at a mass concentration nominally 50 µg/g. The procedures should work with other nanoparticles that carry a negative surface charge or zeta potential. Moreover, these procedures can also be applied to positively charged or neutral nanoparticles with some modification. Each procedure may require some optimization by the user in order to obtain satisfactory deposition density and to minimize artifacts such as aggregate formation on the substrate or build up of organic films resulting from additives that might be present in the solution phase.  This protocol is intended to be an evolving document; addenda that address material-class-specific sample preparation and analysis issues will be appended as available.  Some suggested references for TEM analysis are provided at the end of this document (1-3).

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