In the Laboratory of bioanalytical instrumentation at Masaryk University in Brno, we focus on the development of mass spectrometry (MS) instrumentation and methods for faster and more sensitive bioanalysis. The object of our interest ranges from small organic molecules to metal nanoparticles. This contribution will attempt to give a historical overview of the projects we were involved in. A prerequisite for the instrumental development was mastering the time of flight (TOF) mass analyzer construction; the first apparatus was built for fundamental studies of matrix-assisted laser desorption/ionization (MALDI) using 193 nm laser photodissociation.1 An important research direction was the coupling of microcolumn separations to the TOF mass analyzer; a variety of interfaces were built for in-line and off-line capillary electrophoresis (CE)-MALDI MS and MS/MS analyses of peptide mixtures. Unmatched speed of the TOF mass analyzer along with a kHz laser and a fast galvanometer optical scanner allowed simultaneous detection of multiple separation eluents with a single mass spectrometer.2 A similar idea helped to achieve acquisition rates above 100 pixels per second in MALDI MS imaging experiments recently.3 Another research area was the development of two original techniques of sample introduction for atomic mass spectrometry; combined bioanalysis and elemental analysis hold promising potential in the field of metallomics. Examples involve simultaneous MALDI, electrospray (ESI), and inductively coupled plasma (ICP) MS detection of separated metallothioneins and a thin layer (TLC)-ICP MS of cobalamins or selenium species. One of the sample introduction techniques was employed for the characterization of nanoparticles; it allows the introduction of intact gold nanoparticles into inductively coupled plasma operated in single-particle mode.4 Current research includes multimodal imaging of 3D cell aggregates,5 MS imaging of double bond positional isomers,6 nanoparticle tags for sensitive MS imaging, and a new arrangement for ionization of volatile organic compounds.7 References 1 Preisler, J.; Yeung, E. S. Anal. Chem. 1997, 69, 4390-4398. 2 Preisler, J.; Hu, P.; Rejtar, T.; Moskovets, E.; Karger, B. L. Anal. Chem. 2002, 74, 17-25. 3 Bednarik, A.; Machalkova, M.; Moskovets, E.; Coufalikova, K.; Krasensky, P.; Houska, P.; Kroupa, J.; Navratilova, J.; Smarda, J.; Preisler, J. J. Amer. Soc. Mass Spectrom. 2019, 30, 289-298. 4 Benesova, I.; Dlabkova, K.; Zelenak, F.; Vaculovic, T.; Kanicky, V.; Preisler, J. Anal. Chem. 2016, 88, 2576-2582. 5 Machalkova, M.; Pavlatovska, B.; Michalek, J.; Pruska, A.; Stepka, K.; Necasova, T.; Radaszkiewicz, K. A.; Kozubek, M.; Smarda, J.; Preisler, J.; Navratilova, J. Anal. Chem. 2019, 91, 13475-13484. 6 Bednarik, A.; Preisler, J.; Bezdekova, D.; Machalkova, M.; Hendrych, M.; Navratilova, J.; Knopfova, L.; Moskovets, E.; Soltwisch, J.; Dreisewerd, K. Anal. Chem. 2020, 92, 6245-6250. 7 Bednarik, A.; Prysiazhnyi, V.; Preisler, J. Anal. Chem. 2021, 93, 9445-9453.