Microorganisms have been used by humans for thousands of years in the production of beer, wine, pickles, and milk products. They are also used in the pharmaceutical industry in the production of vaccines, enzymes and antibiotics, and mining for leaching some metals from mineral deposits and waste materials. The use of microorganisms is an interesting approach in the field of bioanalytics. This chapter focuses on the application of microorganisms and other natural materials of plant and animal origin as biosorbents for the preconcentration, separation, and speciation studies of metal ions in analytical chemistry. We describe the mechanisms of metal binding, factors influencing the biosorption process, and methods used for the evaluation of sorption performance of the biomass. Batch and flow extraction techniques using free and immobilized cells, taken from literature and our research results, are presented and discussed. Examples of the latest analytical methodologies based on biosorption of metals and their determination by spectrometric methods in environmental and biological samples are given.