IntroductionIdentification of bacteria by mass spectrometry has revolutionized microbiology in routine diagnostic laboratories within the last years. Many different techniques of mass spectrometry have been used in basic protein research approaches. Procedures such as ESI-ToF (electron spray injection time-of-flight) and SELDI-ToF (surface-enhanced laser desorption ionization time-of-flight) mass spectrometry, or QMSs (quadrupole mass spectrometry) have been developed over the last two decades. But only MALDI-ToF (matrix-assisted laser desorption ionization timeof-flight) mass spectrometry was pushed forward as a simple-to-use, reliable, and cheap technology and succeeded in the routine lab.Mass spectrometry was used already in the 1970s as a new method to identify bacteria [1]. These early papers were mainly published in specialized journals for mass spectrometry and were not recognized by the broader scientific community.To analyze a macromolecule by mass spectrometry, the protein has to be ionized and volatilized by a laser shot. Ionization of molecules by a laser beam includes the risk of the protein breaking down into fragments and losing its tertiary structure. This problem was solved in 1985 by two German scientists, Franz Hillenkamp and Michael Karas. They reported a method that uses small organic compounds as a matrix, protecting the molecules from this damage [2]. In the same year, Koichi Tanaka from Japan presented data where he used a mixture of glycerol and fine metal powder as a matrix. With these tools, he was able to ionize substances with a laser without structural losses. He published the method as soft laser desorption (SLD) after he had filed a patent application [3]. In 2002, Koichi Tanaka was awarded the Nobel Prize in Chemistry, together with John Bennett Fenn and Kurt Wüthrich (the latter for work in NMR spectroscopy), for his invention. Franz Hillenkamp was not considered in the distinction, although he was the first one to publish the method. He had already expanded the application of the new method to a large variety of molecules and higher masses in the year of Tanaka's first publication [4].Modern Techniques for Pathogen Detection, First Edition. Edited by Jürgen Popp and Michael Bauer.