Optical probes for monitoring, imaging, and sensing of pH are of great interest for the scientific community as pH is a crucial marker for many processes in biotechnology, biology, medical diagnostics, biomedical research, and material corrosion. Thereby, optical pH sensors based on fluorescence have attracted interest in particular as fluorescence offers a high sensitivity down to the single molecule level, can be read out with relatively simple and readily miniaturized instrumentation, and allows online in situ measurements. Also the versatility ranging from molecular and nanosensor formats to planar optodes and fiber-optic sensors, and the non-invasive, non-destructive, and contactless nature of the measurement are application-friendly features. The information content, which is offered by a fluorescence intensity-based sensor, is usually unspecific and limited on the presence or the absence of the chromophore or analyte and can additionally be hampered by fluctuation of the excitation light intensity and changes in fluorophore concentration, e.g., due to photobleaching. Therefore, many fluorescence sensors are utilized in referenced systems, which enable twowavelength ratiometric measurements of the fluorescence intensity by the introduction of an analyte-inert reference with a spectrally distinguishable emission. This work presents the rational design of a versatile, modular, multi-component-based platform for ratiometric optical analyte sensing that can be simply adapted to different formats and measurement geometries. Therefore, readily available analyte-responsive fluorescent boron-dipyrromethene (BODIPY) dyes and near infrared (NIR)-excitable multicolour-Partikelgrößen wurden dafür via Transmissionselektronenmikroskopie (TEM) und Kleinwinkel-Röntgenstreuung (SAXS) bestimmt. Neben der Partikelgröße konnten durch die TEM-Messungen auch Informationen über die Kristallphasen der Nanopartikel erhalten werden. Neben der Erfassung des Partikelwachstums wurde die UCL der UCNPs für die ratiometrische Sensorplattform als Nanolampe und gleichzeitig als Referenzsignal verwendet. Die blaue Upconversion(UC)-Emission der NaYF 4 :Yb 3+ /Tm 3+ UCNPs wurde dabei zur Anregung der pH-sensitiven BODIPY-Farbstoffe verwendet, während die rote UC-Emission als inertes Referenzsignal verwendet wurde. Die Berechnung des Verhältnisses der Emissionsintensitäten der grünen Fluoreszenz des Farbstoffs und der roten UC-Emission des Partikels ermöglicht eine Bestimmung des pH-Werts. Das Potenzial dieser Strategie zur Erfassung des pH-Werts wurde beispielhaft für die Bestimmung der zeitabhängigen Änderungen des pH-Werts einer metabolisierenden Escherichia coli (E. coli)-Suspension gezeigt.

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We investigated the growth of β-phase NaYF4:Yb3+,Er3+ upconversion nanoparticles synthesized by the thermal decomposition method using a combination of in situ and offline analytical methods for determining the application-relevant optical properties, size, crystal phase, and chemical composition. This included in situ steady state luminescence in combination with offline time-resolved luminescence spectroscopy as well as small-angle X-ray scattering (SAXS) transmission electron microscopy (TEM), X-ray diffraction analysis (XRD), and inductively coupled plasma optical emission spectrometry (ICP-OES). For assessing the suitability of our optical monitoring approach, the in situ-collected spectroscopic data, which reveal the luminescence evolution during nanocrystal synthesis, were compared to measurements done after cooling of the reaction mixture of the as-synthesized particles. The excellent correlation of the in situ and time-resolved upconversion luminescence with the nanoparticle sizes determined during the course of the reaction provides important insights into the various stages of nanoparticle growth. This study highlights the capability of in situ luminescence monitoring to control the efficiency of UCNP synthesis, particularly the reaction times at elevated temperatures and the particle quality in terms of size, shape, and crystal structure, as well as luminescence lifetime and upconversion quantum yield.

Singlet oxygen can severely damage biological tissue, which is exploited in photodynamic therapy (PDT). In PDT, the effective range is limited by the distribution of the photosensitizer (PS) and the illuminated area. However, no distinction is made between healthy and pathological tissue, which can cause undesired damage. This encouraged us to exploit the more acidic pH of cancerous tissue and design pH-controllable singlet oxygen-generating boron-dipyrromethene (BODIPY) dyes. A pH sensitivity of the dyes is achieved by the introduction of an electronically decoupled, photoinduced electron transfer (PET)-capable subunit in meso-position of the BODIPY core. To favor triplet-state formation as required for singlet oxygen generation, iodine substituents were introduced at the chromophore core. The resulting pH-controlled singlet oxygen-generating dyes with pK a values in the physiological range were subsequently assessed regarding their potential as pH-controlled PS for PDT. Using HeLa cells, we could successfully demonstrate markedly different pH-dependent cytotoxicities upon illumination.

We report on the temperature-and structural-dependent optical properties and photophysics of a set of boron dipyrromethene (BODIPY) dyes with different substitution patterns of their meso-aryl subunit. Single-crystal Xray diffraction analysis of the compounds enabled a classification of the dyes into a sterically hindered and a unhindered group. The steric hindrance refers to a blocked rotational motion of the aryl subunit around the bond connecting this moiety to the meso-position of the BODIPY core. The energy barriers related to this rotation were simulated by DFT calculations. As follows from the relatively low rotational barrier calculated to about 17 kcal/mol, a free rotation is only possible for sterically unhindered compounds. Rotational barriers of more than 40 kcal/mol determined for the sterically hindered compounds suggest an effective freezing of the rotational motion in these molecules. With the aid of temperature-dependent spectroscopic measurements, we could show that the ability to rotate directly affects the optical properties of our set of BODIPY dyes. This accounts for the strong temperature dependence of the fluorescence of the sterically unhindered compounds which show a drastic decrease in fluorescence quantum yield and a significant shortening in fluorescence lifetime upon heating. The optical properties of the sterically hindered compounds, however, are barely affected by temperature. Our results suggest a nonradiative deactivation of the first excited singlet state of the sterically unhindered compounds caused by a conical intersection of the potential energy surfaces of the ground and first excited state which is accessible by rotation of the meso-subunit. This is in good agreement with previously reported deactivation mechanisms. In addition, our results suggest the presence of a second nonradiative depopulation pathway of the first excited singlet state which is particularly relevant for the sterically hindered compounds.