In order to provide a smooth painting surface, a ground layer is applied onto the wooden panel. In northern Europe chalk forms a key constituent of this layer and the tiny nannoplankton from which it is formed can be examined to identify where the chalk might have been mined – this can help establish where the painting could have been produced.

Several marker species of nannoplankton were identified in a small sample taken from this painting. Based on which chalk quarries were active during the sixteenth century, this analysis suggests that the chalk could have been sourced from Kent and Norfolk in England, and northern Europe including Germany, Poland and parts of southern Russia.