Non-stationary variogram models for geostatistical sampling optimisation: An empirical investigation using elevation data

A problem with use of the geostatistical Kriging error for optimal sampling design is that the design does not adapt locally to the character of spatial variation. This is because a stationary variogram or covariance function is a parameter of the geostatistical model. The objective of this paper was to investigate the utility of non-stationary geostatistics for optimal sampling design. First, a contour data set of Wiltshire was split into 25 equal sub-regions and a local variogram was predicted for each. These variograms were fitted with models and the coefficients used in Kriging to select optimal sample spacings for each sub-region. Large differences existed between the designs for the whole region (based on the global variogram) and for the sub-regions (based on the local variograms). Second, a segmentation approach was used to divide a digital terrain model into separate segments. Segment-based variograms were predicted and fitted with models. Optimal sample spacings were then determined for the whole region and for the sub-regions. It was demonstrated that the global design was inadequate, grossly over-sampling some segments while under-sampling others.

Systematic random sampling of the comet assay

The comet assay is a technique used to quantify DNA damage and repair at a cellular level. In the assay, cells are embedded in agarose and the cellular content is stripped away leaving only the DNA trapped in an agarose cavity which can then be electrophoresed. The damaged DNA can enter the agarose and migrate while the undamaged DNA cannot and is retained. DNA damage is measured as the proportion of the migratory ‘tail’ DNA compared to the total DNA in the cell. The fundamental basis of these arbitrary values is obtained in the comet acquisition phase using fluorescence microscopy with a stoichiometric stain in tandem with image analysis software. Current methods deployed in such an acquisition are expected to be both objectively and randomly obtained. In this paper we examine the ‘randomness’ of the acquisition phase and suggest an alternative method that offers both objective and unbiased comet selection. In order to achieve this, we have adopted a survey sampling approach widely used in stereology, which offers a method of systematic random sampling (SRS). This is desirable as it offers an impartial and reproducible method of comet analysis that can be used both manually or automated. By making use of an unbiased sampling frame and using microscope verniers, we are able to increase the precision of estimates of DNA damage. Results obtained from a multiple-user pooled variation experiment showed that the SRS technique attained a lower variability than that of the traditional approach. The analysis of a single user with repetition experiment showed greater individual variances while not being detrimental to overall averages. This would suggest that the SRS method offers a better reflection of DNA damage for a given slide and also offers better user reproducibility.