The value assignment for properties of six certified reference materials (ERM-AD623a–f), each containing a plasmid DNA solution ranging from 1 million to 10 copies per μL, by using digital PCR (dPCR) with the BioMark™ HD System (Fluidigm) has been verified by applying droplet digital PCR (ddPCR) using the QX100 system (Bio-Rad). One of the critical factors in the measurement of copy number concentrations by digital PCR is the partition volume. Therefore, we determined the average droplet volume by optical microscopy, revealing an average droplet volume that is 8 % smaller than the droplet volume used as the defined parameter in the QuantaSoft software version 1.3.2.0 (Bio-Rad) to calculate the copy number concentration. This observation explains why copy number concentrations estimated with ddPCR and using an average droplet volume predefined in the QuantaSoft software were systematically lower than those measured by dPCR, creating a significant bias between the values obtained by these two techniques. The difference was not significant anymore when the measured droplet volume of 0.834 nL was used to estimate copy number concentrations. A new version of QuantaSoft software (version 1.6.6.0320), which has since been released with Bio-Rad’s new QX200 systems and QX100 upgrades, uses a droplet volume of 0.85 nL as a defined parameter to calculate copy number concentration.Graphical AbstractMonolayer of droplets generated by the droplet generator and observed under an optical microscope

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A new certified reference material (CRM) for size and shape analysis of elongated nanoparticles has been developed by the European Commission’s Joint Research Centre. The CRM consists of titanium dioxide nanorods dispersed in 1-butanol, was coded ERM-FD103 and has been certified for different electron microscopy–based operationally defined measurands such as the modal and median values of the particle number-weighted distributions of the minimum and maximum Feret diameter, the maximum inscribed circle diameter, the area-equivalent circular diameter and the aspect ratio. The nanorods have nominal dimensions of 15 nm in width and 55 nm in length. Homogeneity and stability measurements were performed using transmission electron microscopy. The relative standard uncertainty for homogeneity ranged from 0.3 to 1.7%. No significant instability was detected for a shelf life of 18 months and a storage temperature of 18 °C. The certified values have been determined from the results of an interlaboratory comparison in which qualified expert laboratories participated with scanning and transmission electron microscopy. The certified values are traceable to the unit of length in the International System of Units, the metre, and the relative expanded uncertainties (confidence level of approximately 95%) range from 4 to 6%. These properties allow the CRM to be used for quality assurance and calibration of electron microscopy methods for nanoparticle size and shape analysis in ranges relevant for the implementation of EU legislation related to nanomaterials. The presented study discusses the purpose and results of the different steps that were followed to turn an industrially relevant raw titanium dioxide nanorod material into a fit-for-purpose CRM. Graphical abstract

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