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Health-related monitoring of bioaerosol exposures in the workplace should ideally be carried out using size-selective personal samplers that separate the aerosol into biologically relevant size fractions and allow both quantification and identification of the microorganisms present in each fraction. As a first stage in the development of personal bioaerosol samplers a number of collection substrates were assessed for their ability to maintain the viability of the collected microorganisms, so that subsequent culturing and species identification may be carried out. The substrates were tested with bioaerosols of varying robustness, consisting of Saccharomyces cerevisiae cells, Escherichia coli cells, and Penicillium expansum spores, aerosolized under controlled environmental conditions. The survival of microorganisms on each test substrate, assessed on the basis of the culturable fractions of cells recovered, was compared with that of microorganisms collected in a reference glass cyclone sampler. These bioefficiency tests identified the substrate combinations with the potential to fulfill personal sampler design criteria. The substrates were then subjected to further development to evaluate and optimize their particle size selection characteristics. The outcome of this work is two prototype personal bioaerosol samplers in which size-selective substrates are adapted for use in existing designs of personal inhalable sampler. This offers an effective and low-cost solution to personal monitoring of bioaerosol exposures in the workplace.