Epidemiologic research has shown increases in adverse cardiovascular and respiratory outcomes in relation to mass concentrations of particulate matter (PM) < or = 2.5 or < or = 10 microm in diameter (PM2.5, PM10, respectively). In a companion article [Delfino RJ, Sioutas C, Malik S. 2005. Environ Health Perspect 113(8):934-946]), we discuss epidemiologic evidence pointing to underlying components linked to fossil fuel combustion. The causal components driving the PM associations remain to be identified, but emerging evidence on particle size and chemistry has led to some clues. There is sufficient reason to believe that ultrafine particles < 0.1 microm (UFPs) are important because when compared with larger particles, they have order of magnitudes higher particle number concentration and surface area, and larger concentrations of adsorbed or condensed toxic air pollutants (oxidant gases, organic compounds, transition metals) per unit mass. This is supported by evidence of significantly higher in vitro redox activity by UFPs than by larger PM. Although epidemiologic research is needed, exposure assessment issues for UFPs are complex and need to be considered before undertaking investigations of UFP health effects. These issues include high spatial variability, indoor sources, variable infiltration of UFPs from a variety of outside sources, and meteorologic factors leading to high seasonal variability in concentration and composition, including volatility. To address these issues, investigators need to develop as well as validate the analytic technologies required to characterize the physical/chemical nature of UFPs in various environments. In the present review, we provide a detailed discussion of key characteristics of UFPs, their sources and formation mechanisms, and methodologic approaches to assessing population exposures.