@misc{WessigJohnSperlichetal.2020, author = {Wessig, Pablo and John, Leonard and Sperlich, Eric and Kelling, Alexandra}, title = {Sulfur tuning of [1,3]-dioxolo[4.5-f]benzodioxole (DBD) fluorescent dyes}, series = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, journal = {Zweitver{\"o}ffentlichungen der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, number = {3}, issn = {1866-8372}, doi = {10.25932/publishup-56624}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-566241}, pages = {15}, year = {2020}, abstract = {The replacement of oxygen by sulfur atoms of [1,3]-dioxolo[4.5-f]benzodioxole (DBD) fluorescent dyes is an efficient way to adjust the photophysical properties (sulfur tuning). While previously developed S-4-DBD dyes exhibit considerably red-shifted absorption and emission wavelength, the heavy atom effect of four sulfur atoms cause low fluorescence quantum yields and short fluorescence lifetimes. Herein, we demonstrate that the replacement of less than four sulfur atoms (S-1-DBD, 1,2-S-2-DBD, and 1,4-S-2-DBD dyes) permits a fine-tuning of the photophysical properties. In some cases, a similar influence on the wavelength without the detrimental effect on the quantum yields and lifetimes is observed. Furthermore, the synthetic accessibility of S-1- and S-2-DBD dyes is improved, compared with S-4-DBD dyes. For coupling with biomolecules a series of reactive derivatives of the new dyes were developed (azides, OSu esters, alkynes, maleimides).}, language = {en} } @article{HaubitzJohnFreyseetal.2020, author = {Haubitz, Toni and John, Leonard and Freyse, Daniel and Wessig, Pablo and Kumke, Michael Uwe}, title = {Investigating the Sulfur "Twist" on the Photophysics of DBD Dyes}, series = {The journal of physical chemistry : A, Molecules, spectroscopy, kinetics, environment \& general theory}, volume = {124}, journal = {The journal of physical chemistry : A, Molecules, spectroscopy, kinetics, environment \& general theory}, number = {22}, publisher = {American Chemical Society}, address = {Washington}, issn = {1089-5639}, doi = {10.1021/acs.jpca.0c01880}, pages = {4345 -- 4353}, year = {2020}, abstract = {The so-called DBD ([1,3]dioxolo[4,5-f][1,3]benzodioxole) dyes are a new class of fluorescent dyes, with tunable photophysical properties like absorption, fluorescence lifetime, and Stokes shift. With the development of sulfur based DBDs, this dye class is extended even further for possible applications in spectroscopy and microscopy. In this paper we are investigating the basic photophysical properties and their implications for future applications for S-4-DBD as well as O-4-DBD. On the basis of time-resolved laser fluorescence spectroscopy, transient absorption spectroscopy, and UV/vis-spectroscopy, we determined the rate constants of the radiative and nonradiative deactivation processes as well as the energy of respective electronic states involved in the electronic deactivation of S-4-DBD and of O-4-DBD. For S-4-DBD we unraveled the triplet formation with intersystem crossing quantum yields of up to 80\%. By TD-DFT calculations we estimated a triplet energy of around 13500-14700 cm(-1) depending on the DBD dye and solvent. Through solvent dependent measurements, we found quadrupole moments in the range of 2 B.}, language = {en} } @article{WessigJohnSperlichetal.2020, author = {Wessig, Pablo and John, Leonard and Sperlich, Eric and Kelling, Alexandra}, title = {Sulfur tuning of [1,3]-dioxolo[4.5-f]benzodioxole (DBD) fluorescent dyes}, series = {European journal of organic chemistry}, volume = {2021}, journal = {European journal of organic chemistry}, number = {3}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1434-193X}, doi = {10.1002/ejoc.202001418}, pages = {499 -- 511}, year = {2020}, abstract = {The replacement of oxygen by sulfur atoms of [1,3]-dioxolo[4.5-f]benzodioxole (DBD) fluorescent dyes is an efficient way to adjust the photophysical properties (sulfur tuning). While previously developed S-4-DBD dyes exhibit considerably red-shifted absorption and emission wavelength, the heavy atom effect of four sulfur atoms cause low fluorescence quantum yields and short fluorescence lifetimes. Herein, we demonstrate that the replacement of less than four sulfur atoms (S-1-DBD, 1,2-S-2-DBD, and 1,4-S-2-DBD dyes) permits a fine-tuning of the photophysical properties. In some cases, a similar influence on the wavelength without the detrimental effect on the quantum yields and lifetimes is observed. Furthermore, the synthetic accessibility of S-1- and S-2-DBD dyes is improved, compared with S-4-DBD dyes. For coupling with biomolecules a series of reactive derivatives of the new dyes were developed (azides, OSu esters, alkynes, maleimides).}, language = {en} } @article{WessigJohnMertens2018, author = {Wessig, Pablo and John, Leonard and Mertens, Monique}, title = {Extending the Class of [1,3]-Dioxolo[4.5-f]benzodioxole (DBD) Fluorescent Dyes}, series = {European journal of organic chemistry}, volume = {2018}, journal = {European journal of organic chemistry}, number = {14}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1434-193X}, doi = {10.1002/ejoc.201800002}, pages = {1674 -- 1681}, year = {2018}, abstract = {Synthetic routes to a collection of new fluorescent dyes are described, which are based on the [1,3]-dioxolo[4.5-f]benzodioxole (DBD) core. By introducing different electron withdrawing groups in 4- and 8-position of the DBD moiety the emission wavelength could be adjusted over a large spectral range from blue to orange light.}, language = {en} } @article{BuechnerJohnMertensetal.2018, author = {B{\"u}chner, D{\"o}rthe and John, Leonard and Mertens, Monique and Wessig, Pablo}, title = {Detection of dsDNA with [1,3]Dioxolo[4,5-f]benzodioxol (DBD) Dyes}, series = {Chemistry - a European journal}, volume = {24}, journal = {Chemistry - a European journal}, number = {60}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0947-6539}, doi = {10.1002/chem.201804057}, pages = {16183 -- 16190}, year = {2018}, abstract = {DBD fluorescent dyes have proven to be useful in numerous applications. To widen the range of biological applications, we propose three different types of DBD molecules that have been modified in such a way that DNA interaction becomes probable. After the successful synthesis of all three compounds, we tested their fluorescent properties and their DNA binding abilities. Two of the three probes exhibit an interaction with dsDNA with subsequent fluorescence enhancement. The determined binding constants of the two new DNA dyes are comparable to other minorgroove-binding dyes. Their large Stokes shifts and their long fluorescent lifetimes are outstanding features of these dyes.}, language = {en} } @phdthesis{John2021, author = {John, Leonard}, title = {Neuartige DBD-Fluoreszenzfarbstoffe}, doi = {10.25932/publishup-51048}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-510487}, school = {Universit{\"a}t Potsdam}, pages = {257}, year = {2021}, abstract = {Zusammenfassung zur Dissertation „Neuartige DBD-Fluoreszenzfarbstoffe: Synthese, Untersuchungen und Anwendungen" von Leonard John In dieser Arbeit konnten auf Basis der etablierten [1,3]-Dioxolo[4,5-f][1,3]benzodioxol (DBD) Fluoreszenzfarbstoffe zwei neue Konzepte zur Darstellung unsymmetrisch funktionalisierter DBD-Fluorophore entwickelt werden. Die Variation der elektronenziehenden Reste f{\"u}hrte zu einer Erweiterung des Farbspektrums an DBD-Fluorophoren, wobei alle weiteren spektroskopischen Parameter (Fluoreszenzlebenszeit, -quantenausbeute und STOKES-Verschiebung) unver{\"a}ndert hohe Werte aufweisen. Neben der Variation der elektronenziehenden Reste wurde das "pi"-System des DBD-Farbstoffs mit der Einf{\"u}hrung von Stilben-, und Tolan-Derivaten vergr{\"o}ßert. Stilben-Derivate zeigten {\"a}hnlich gute spektroskopische Eigenschaften wie die bereits etablierten DBD-Farbstoffe. Fluorophore mit langwelliger Emission sind auf Grund der großen Gewebe-Eindringtiefe besonders interessant f{\"u}r biologische Anwendungen. Da der langwelligste Vertreter der O4-DBD-Farbstoffe in polaren Medien nur schwer l{\"o}slich ist, wurde ein Weg zur Einf{\"u}hrung l{\"o}slichkeitsvermittelnder Gruppen gesucht. Hierbei fiel die Wahl auf eine Carbons{\"a}ure-Gruppe zur Steigerung der Hydrophilie. Eine von vier untersuchten Methoden erwies sich als zielf{\"u}hrend, sodass das gew{\"u}nschte Molek{\"u}l isoliert werden konnte. Eine erh{\"o}hte Wasserl{\"o}slichkeit wurde allerdings nicht beobachtet. Zur Erforschung von Fettstoffwechselkrankheiten wie der ALZHEIMER-Krankheit werden fluoreszenzmarkierte Lipide ben{\"o}tigt. Um unterschiedliche Bereiche einer Membran zu untersuchen, war das Ziel, den Fluorophor an unterschiedlichen Stellen innerhalb der Fetts{\"a}ure zu lokalisieren. Hierbei sollte die Gesamtkettenl{\"a}nge des DBD-Lipids einer C18-Kette, analog der Stearins{\"a}ure, entsprechen. Durch die stufenweise Einf{\"u}hrung der Reste gelang es, drei DBD-Lipide herzustellen, wobei sich der Fluorophor an unterschiedlichen Positionen innerhalb der Kette befindet. Die photophysikalischen Eigenschaften der Lipide weichen nur marginal von denen der reinen Fluorophore ab. Eine Einlagerung in giant unilamellar vesicles (GUVs) konnte f{\"u}r zwei Derivate beobachtet werden, wobei keine dom{\"a}nenspezifisch war. Ein weiteres Ziel dieser Arbeit war es, die vier Sauerstoffatome im DBD-Grundk{\"o}rper stufenweise durch Schwefelatome zu ersetzen und die Ringgr{\"o}ßen des DBD-Fluorophors zu variieren. F{\"u}r die Ringgr{\"o}ße zeigte der 1,2-S2-DBD mit jeweils zwei F{\"u}nfringen die besten spektroskopischen Eigenschaften. Durch die Synthese von zwei weiteren schwefelhaltigen DBD-Grundk{\"o}rpern (S1- und 1,4-S2-DBD) konnten insgesamt drei neue Farbstoffklassen zug{\"a}nglich gemacht werden. F{\"u}r alle neuen Chromophore wurden elektronenziehende Reste (Aldehyd, Acyl, Ester, Carboxy) eingef{\"u}hrt und die jeweiligen Derivate spektroskopisch untersucht. Mit steigender Anzahl an Schwefel-Atomen im Grundk{\"o}rper zeigt sich eine bathochrome Verschiebung der Emission, wobei die Werte f{\"u}r die Fluoreszenzlebenszeit- und -quantenausbeute abnehmen. Die optimalen spektroskopischen Eigenschaften aus langwelliger Emission, hoher Fluoreszenzlebenszeit und -quantenausbeute zeigt das 1,4-S2-Dialdehyd-Derivat. F{\"u}r die S1- und 1,2-S2-Dialdehyd- Derivate wurden Konzepte entwickelt, um bioreaktive Reste (Alkin, HOSu, Maleimid) einzuf{\"u}hren und die Fluorophore in biologischen Systemen anwenden zu k{\"o}nnen.}, language = {de} } @misc{SchwarzeRiemerMuelleretal.2019, author = {Schwarze, Thomas and Riemer, Janine and M{\"u}ller, Holger and John, Leonard and Holdt, Hans-J{\"u}rgen and Wessig, Pablo}, title = {Na+ Selective Fluorescent Tools Based on Fluorescence Intensity Enhancements, Lifetime Changes, and on a Ratiometric Response}, series = {Postprints der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, journal = {Postprints der Universit{\"a}t Potsdam : Mathematisch-Naturwissenschaftliche Reihe}, number = {1136}, issn = {1866-8372}, doi = {10.25932/publishup-43748}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-437482}, pages = {13}, year = {2019}, abstract = {Over the years, we developed highly selective fluorescent probes for K+ in water, which show K+-induced fluorescence intensity enhancements, lifetime changes, or a ratiometric behavior at two emission wavelengths (cf. Scheme 1, K1-K4). In this paper, we introduce selective fluorescent probes for Na+ in water, which also show Na+ induced signal changes, which are analyzed by diverse fluorescence techniques. Initially, we synthesized the fluorescent probes 2, 4, 5, 6 and 10 for a fluorescence analysis by intensity enhancements at one wavelength by varying the Na+ responsive ionophore unit and the fluorophore moiety to adjust different K-d values for an intra- or extracellular Na+ analysis. Thus, we found that 2, 4 and 5 are Na+ selective fluorescent tools, which are able to measure physiologically important Na+ levels at wavelengths higher than 500 nm. Secondly, we developed the fluorescent probes 7 and 8 to analyze precise Na+ levels by fluorescence lifetime changes. Herein, only 8 (K-d=106 mm) is a capable fluorescent tool to measure Na+ levels in blood samples by lifetime changes. Finally, the fluorescent probe 9 was designed to show a Na+ induced ratiometric fluorescence behavior at two emission wavelengths. As desired, 9 (K-d=78 mm) showed a ratiometric fluorescence response towards Na+ ions and is a suitable tool to measure physiologically relevant Na+ levels by the intensity change of two emission wavelengths at 404 nm and 492 nm.}, language = {en} } @article{HaubitzJohnWessigetal.2019, author = {Haubitz, Toni and John, Leonard and Wessig, Pablo and Kumke, Michael Uwe}, title = {Photophysics of Acyl- and Ester-DBD Dyes}, series = {the journal of physical chemistry : A, Molecules, spectroscopy, kinetics, environment \& general theory}, volume = {123}, journal = {the journal of physical chemistry : A, Molecules, spectroscopy, kinetics, environment \& general theory}, number = {22}, publisher = {American Chemical Society}, address = {Washington}, issn = {1089-5639}, doi = {10.1021/acs.jpca.9b02973}, pages = {4717 -- 4726}, year = {2019}, abstract = {A new generation of wavelength-tunable, fluorescent dyes, so-called DBD ([1,3]dioxolo[4,5-f][1,3]benzodioxole) dyes, were developed a few years ago, and they showed great potential as probes, for example, for fluorescence microscopy. However, their photophysics is not fully explored and leaves open questions regarding their large fluorescence Stokes shifts and sensitivity to solvent conditions of differently substituted DBD dyes. To improve the understanding of the influence of the substitution pattern of the DBD dyes on their respective photophysics, transient absorption spectroscopy (TAS) was used, that is, a pump-probe experiment on the femtosecond timescale. TAS allows measurements of excited states, ground state recovery, solvent relaxation, and fluorescence properties on time scales of up to several nanoseconds. Two different DBD dye samples were investigated: aryl- and ester-substituted DBD dyes. Experiments were carried out in solvents with different polarities using different excitation energies and at different viscosities. Based on the experimental data and theoretical calculations, we were able to determine the conformational changes of the molecule due to electronic excitation and were able to investigate solvent relaxation processes for both types of DBD dyes. By generalizing the theory for quadrupole-induced solvent relaxation developed by Togashi et al., we derived quadrupole moments of both molecules in the ground and excited state. Our data showed differences in the binding of polar solvent molecules to the dyes depending on the substituent on the DBD dye. In the case of water as the solvent, an additional efficient quenching process in the electronically excited state was revealed, which was indicated by the observation of solvated electrons in the TAS signals.}, language = {en} } @article{SchwarzeRiemerMuelleretal.2019, author = {Schwarze, Thomas and Riemer, Janine and M{\"u}ller, Holger and John, Leonard and Holdt, Hans-J{\"u}rgen and Wessig, Pablo}, title = {Na+ Selective Fluorescent Tools Based on Fluorescence Intensity Enhancements, Lifetime Changes, and on a Ratiometric Response}, series = {Chemistry - a European journal}, volume = {25}, journal = {Chemistry - a European journal}, number = {53}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0947-6539}, doi = {10.1002/chem.201902536}, pages = {12412 -- 12422}, year = {2019}, abstract = {Over the years, we developed highly selective fluorescent probes for K+ in water, which show K+-induced fluorescence intensity enhancements, lifetime changes, or a ratiometric behavior at two emission wavelengths (cf. Scheme 1, K1-K4). In this paper, we introduce selective fluorescent probes for Na+ in water, which also show Na+ induced signal changes, which are analyzed by diverse fluorescence techniques. Initially, we synthesized the fluorescent probes 2, 4, 5, 6 and 10 for a fluorescence analysis by intensity enhancements at one wavelength by varying the Na+ responsive ionophore unit and the fluorophore moiety to adjust different K-d values for an intra- or extracellular Na+ analysis. Thus, we found that 2, 4 and 5 are Na+ selective fluorescent tools, which are able to measure physiologically important Na+ levels at wavelengths higher than 500 nm. Secondly, we developed the fluorescent probes 7 and 8 to analyze precise Na+ levels by fluorescence lifetime changes. Herein, only 8 (K-d=106 mm) is a capable fluorescent tool to measure Na+ levels in blood samples by lifetime changes. Finally, the fluorescent probe 9 was designed to show a Na+ induced ratiometric fluorescence behavior at two emission wavelengths. As desired, 9 (K-d=78 mm) showed a ratiometric fluorescence response towards Na+ ions and is a suitable tool to measure physiologically relevant Na+ levels by the intensity change of two emission wavelengths at 404 nm and 492 nm.}, language = {en} }