raman spectroscopic method for identification of clinically relevant microorganisms growing on solid culture medium routine clinical microbiological identification of pathogenic microorganisms is largely based on nutritional and biochemical tests in the case of severely ill patients the unavoidable time delay associated with such identification procedures can be fatal we present a novel identification method based on confocal raman microspectroscopy with this approach it is possible to obtain raman spectra directly from microbial microcolonies on the solid culture medium which have developed after only 6 h of culturing for the most commonly encountered organisms due to the limited thickness of microcolonies some of the underlying culture medium is sampled together with the bacteria spectra measured at different depths in a microcolony contain different amounts of the medium signal a mathematical routine involving vector algebra is described for the nonsubjective correction of spectra for variable signal contributions of the medium to illustrate the possibilities of our approach for the identification of microorganisms raman spectra were collected from 6h microcolonies of five bacterial strains on solid culture medium the classification results show that confocal raman microspectroscopy has great potential as a powerful new tool in clinical diagnostic microbiology