The hydrogen molecule is the smallest neutral chemical entity and a benchmark system of molecular spectroscopy. The comparison between highly accurate measurements of transition frequencies and level energies with quantum calculations including all known phenomena (relativistic, vacuum polarization and self energy) provides a tool to search for physical phenomena in the realm of the unknown: are there forces beyond the three included in the Standard Model of physics plus gravity [1], are there extra dimensions beyond the 3+1 describing space time [2] ? Comparison of laboratory wavelengths of transitions in hydrogen may be compared with the lines observed during the epoch of the early Universe to verify whether fundamental constants of Nature have varied over cosmological time [3]. These concepts, as well as the precision laboratory experiments and the astronomical observations used for such searches of new physics [4] will be discussed. [1] E.J. Salumbides, J.C.J. Koelemeij, J. Komasa, K. Pachucki, K.S.E. Eikema, W. Ubachs, Bounds on fifth forces from precision measurements on molecules, Phys. Rev. D87, 112008 (2013). [2] E.J. Salumbides, A.N. Schellekens, B. Gato-Rivera, W. Ubachs Constraints on extra dimensions from molecular spectroscopy, New. J. Phys. 17, 033015 (2015). [3] W. Ubachs, J. Bagdonaite, E.J. Salumbides, M.T. Murphy, L. Kaper, Search for a drifting proton-electron mass ratio from H₂, Rev. Mod. Phys. 88, 021003 (2016). [4] W. Ubachs, J.C.J. Koelemeij, K.S.E. Eikema, E.J. Salumbides, Physics beyond the Standard Model from hydrogen spectroscopy, J. Mol. Spectr. 320, 1 (2016).

In the early 2000’s, the work of Cole and Legon,G.C. Cole and A.C. Legon, Chem. Phys. Lett. 369, 31-40 (2003).^, G.C. Cole and A.C. Legon, Chem. Phys. Lett. 400, 414-424 (2004).cZ. Kisiel, P.W. Fowler, and A.C. Legon, J. Chem. Phys. 93, 3054-3062 (1990).d

Owing to its unparalleled structural specificity, rotational spectroscopy is a powerful technique to unambiguously identify and characterize polar molecules. We present here an experimental approach, automated microwave double resonance (AMDOR) spectroscopy, that allows to rapidly determine the rotational constants of such compounds without any a priori knowledge of elemental composition or molecular structure. This task is achieved by acquiring the classical (frequency vs. intensity) broadband spectrum of a molecule using chirped-pulse Fourier transform microwave (FTMW) spectroscopy, and subsequently analyzing it in near-real time using complementary cavity FTMW detection and double resonance. AMDOR measurements provide a unique "barcode" for each compound from which rotational constants can be extracted. Results obtained on the characterization of individual compounds and mixtures will be described.

Innovations in light microscopy have tremendously revolutionized the way researchers study biological systems with subcellular resolution. Although fluorescence microscopy is currently the method of choice for cellular imaging, it faces fundamental limitations for studying the vast number of small biomolecules. This is because relatively bulky fluorescent labels could introduce considerable perturbation to or even completely alter the native functions of vital small biomolecules. Hence, despite their immense functional importance, these small biomolecules remain largely undetectable by fluorescence microscopy. To address this challenge, we have developed a bioorthogonal chemical imaging platform. By coupling stimulated Raman scattering (SRS) microscopy, an emerging nonlinear Raman microscopy technique, with tiny and Raman-active vibrational probes (e.g., alkynes, nitriles and stable isotopes including 2H and 13C), bioorthogonal chemical imaging exhibits superb sensitivity, specificity, multiplicity and biocompatibility for imaging small biomolecules in live systems including tissues and organisms. Exciting biomedical applications such as imaging fatty acid metabolism related to lipotoxicity, glucose uptake and metabolism, drug trafficking, protein synthesis, DNA replication, protein degradation, RNA synthesis and tumor metabolism will be presented. This bioorthogonal chemical imaging platform is compatible with live-cell biology, thus allowing real-time imaging of small-molecule dynamics. Moreover, further chemical and spectroscopic strategies allow for multicolor bioorthogonal chemical imaging, a valuable technique in the era of “omics”. We envision that the coupling of SRS microscopy with vibrational probes would do for small biomolecules what fluorescence microscopy of fluorophores has done for larger molecular species, bringing small molecules under the illumination of modern light microscopy.