The influence of air pollution on epiphytic lichens in Košice city has been studied. We observed differences in number of species as well as lichen abundance at sites close to a steel factory south of the city, the city center, and peripheral parts north of the city. For the city center and sites close to steel factory, lichens more tolerant to pollution from Zone 3-4 were typical. However, on the north periphery of the city (site “Alpinka”) we found even Ramalina fastigiata, a typical member of zone 7, which include lichens very sensitive to air pollution [30]. We demonstrated by chlorophyll analysis of transplanted Hypogymnia physodes, that chlorophyll a degradation (expressed as ratio of OD 435/OD 415) negatively correlates with degree of lichen diversity and abundance at the studied sites. Using EDX-microanalysis we determined amounts of elements in lichen thalli of Lecanora chlarotera, Physcia tenella and bark of the tree Populus tremula (lichen substrate) near U.S. Steel in Košice due to determine the chemical nature of air pollution. Similarly, we analyzed the amount of these pollutants in control lichens Flavoparmelia caperata, Ramalina fastigiata and Physcia aipolia, grown in northern peripheral parts of the city. We demonstrated possibilities to parallel the use of several methodologies in assessment of air pollution by lichens in urban areas with intensive industry.