TU Wien 2 articles published in JoVE Bioengineering Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes Lukas Schrangl1, Janett Göhring2, Florian Kellner2, Johannes B. Huppa2, Gerhard J. Schütz1 1Institute of Applied Physics, TU Wien, 2Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna This article presents a method for spatiotemporal analysis of mobile, single-molecule Förster resonance energy transfer (smFRET)-based probes using widefield fluorescence microscopy. The newly developed software toolkit allows the determination of smFRET time traces of moving probes, including the correct FRET efficiency and the molecular positions, as functions of time. Environment Isolation of Mandibular Gland Reservoir Contents from Bornean 'Exploding Ants' (Formicidae) for Volatilome Analysis by GC-MS and MetaboliteDetector Michaela Hoenigsberger1, Alexey G. Kopchinskiy2, Alexandra Parich1, Karsten Hiller3,4, Alice Laciny5, Herbert Zettel5, Linda B.L. Lim6, Kamariah A. Salim7, Irina S. Druzhinina2, Rainer Schuhmacher1 1Center for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences Vienna (BOKU), Austria, 2Institute of Chemical, Environmental and Biological Engineering, TU Vienna, Austria, 3Department of Computational Biology of Infection Research, Helmholtz Centre for Infection Research, Germany, 4Institute for Biochemistry, Biotechnology and Bioinformatics, University of Technology (TU) Braunschweig, Germany, 52nd Zoological Department, Natural History Museum Vienna, Austria, 6Chemical Sciences, Universiti Brunei Darussalam, Brunei, 7Environmental and Life Sciences, Universiti Brunei Darussalam, Brunei Minor workers of the ant species Colobopsis explodens are dissected to isolate the wax-like content stored in their hypertrophied mandibular gland reservoirs for subsequent solvent-extraction and analysis by gas chromatography-mass spectrometry. The annotation and identification of volatile constituents using the open-source software MetaboliteDetector is also described.