Two formulations of acrylic varnish, with and without either of two dry film biocides-one a mixture of isothiazolinones and benzimidazole derivatives, and the other a carbamate-were tested in vitro for their activity against mixtures of filamentous fungi and cyanobacteria found on sandstone buildings. Growth on filter-paper squares coated with the varnishes was assessed semi-quantitatively by naked eye, quantitatively by image analysis and chemically by measurement of ergosterol and chlorophyll a. The lower solvent content (higher resin) varnish was more inhibitory to cyanobacteria than the higher varnish content, whilst the opposite was true for the fungal inoculum. The carbamate biocide was effective against cyanobacteria, unlike the isothiazolinone mixture, but the latter produced more inhibition of fungal growth. The three assay methods produced generally similar results, although visual observation was obviously the most imprecise. There was an anomaly in the ergosterol measurements, which was considered to be caused by the varying ergosterol content and unequal inhibition of the three fungal genera used in the inoculum. Fusarium sp. was shown to contain higher levels of this membrane component than Cladosporium sp. and Penicillium sp. For this reason, the most appropriate method overall, giving reliable quantitative results, was deemed to be the image analysis.