Near‐Infrared Multipurpose Lanthanide‐Imaging Nanoprobes

Optical imaging plays a growing role in modern biomedical research and clinical applications due to its high sensitivity, superb spatiotemporal resolution and minimal hazards. Lanthanide‐doped nanoparticles (LDNPs), as a classical category of luminescent materials, exhibit promising photostability, near‐infrared (NIR)‐excited frequency up‐/down‐converting capabilities, emission fine‐tuning and multispectral features, which have greatly promoted the endeavors of deeper and clearer diagnostics in complex living conditions. This review focuses on the recent advances of LDNP‐based multipurpose imaging studies using upconversion, downshifting, lifetime, photoacoustic and multimodal nanoprobes in the NIR (650–1000 nm) and the second near‐infrared window (NIR‐II, 1000–1700 nm). The principle and design of various functional, activatable, multiplexing or multimodal lanthanide‐imaging nanoprobes (LINPs) as well as representative biophotonic applications are summarized in detail. In addition, the future perspectives and challenges for facilitating LINPs to clinical translations are discussed.     

Time resolved spectroscopy in laser grade organic dyes

Time resolved spectroscopy is discussed along with the theoretical and experimental methods used to measure the fluorescence lifetimes in various laser grade organic dyes. The role of solvents and their viscosities in the fluorescence life time is also discussed. Different photophysical parameters have been obtained in rigid and non-rigid dye molecules.     

Energy Levels and Band Intensities of Neodymium Organic Acid Complexes

Abstract

The optical absorption spectra of Nd³⁺ ion in glutamic, L-aspartic and maleic acids in UV-VIS and NIR regions have been recorded. From the spectral data thus obtained, various spectroscopic parameters are evaluated. The radiative lifetimes (γ_R) and branching ratios (β) for the excited states of Nd³⁺ ion in three acids have been computed. The relationship between the environment sensitive Judd-Ofelt parameter (ω₂) and the intensities of hypersensitive level (⁴G_5/2) is in good coincidence following the Judd-Ofelt theory.     

Correlation of soft magnetic properties with free volume and medium range ordering in metallic glasses probed by fluctuation microscopy and positron annihilation technique

Amorphous ribbons of different thicknesses of Co_64.5Fe_3.5Si₁₆B₁₄Ni₂ alloy were synthesized using the melt spinning technique by varying wheel speed. The effect of cooling rate on the ribbon thickness and their soft magnetic properties have been studied. The amorphous structure has been characterized in terms of structural free volume and medium range order (MRO) by positron annihilation spectroscopy and fluctuation electron microscopy techniques. Positron lifetime spectra of amorphous samples showed two lifetime components. The first component was found to be correlated with MRO whereas, the second lifetime component was found to be associated with nanovoid type of defects, and the second component was strongly dependent on processing conditions. It could be established that the coercivity of the amorphous samples produced by the rapid solidification technique mainly depends on the defects formed during processing rather than change induced in MRO.     

Effects of Refractive Index and Viscosity on Fluorescence and Anisotropy Decays of Enhanced Cyan and Yellow Fluorescent Proteins

The fluorescence lifetime strongly depends on the immediate environment of the fluorophore. Time-resolved fluorescence measurements of the enhanced forms of ECFP and EYFP in water–glycerol mixtures were performed to quantify the effects of the refractive index and viscosity on the fluorescence lifetimes of these proteins. The experimental data show for ECFP and EYFP two fluorescence lifetime components: one short lifetime of about 1 ns and a longer lifetime of about 3.7 ns of ECFP and for EYFP 3.4. The fluorescence of ECFP is very heterogeneous, which can be explained by the presence of two populations: a conformation (67% present) where the fluorophore is less quenched than in the other conformation (33% present). The fluorescence decay of EYFP is much more homogeneous and the amplitude of the short fluorescence lifetime is about 5%. The fluorescence anisotropy decays show that the rotational correlation time of both proteins scales with increasing viscosity of the solvent similarly as shown earlier for GFP. The rotational correlation times are identical for ECFP and EYFP, which can be expected since both proteins have the same shape and size. The only difference observed is the slightly lower initial anisotropy for ECFP as compared to the one of EYFP.     

Steady-State and Time-Resolved Spectroscopic Characteristics of Mesoionic Oxazolones Solutions

Three new mesoionic oxazolo[3,2-b]pyridazin-2-one derivatives, in different solutions have been investigated by UV-Vis absorption, steady-state and time-resolved fluorescence methods. The effect of substituents on the extension of conjugation of the pi-electrons from mesoionic oxazolone has been evidenced by bathochromic shifts of the absorption and fluorescence maxima positions. The fluorescence decay data could be fitted to single-exponential or double-exponential function. The lifetime values are much higher in aprotic polar solvents and in the case of the derivatives that present an extension of the conjugation of pi-electrons. The properties of the compounds present a solvent dependence, being tested in micellar solutions as potential molecular probe "sensitive" to the environment polarity.     

Probing the Conformational Stability of Two Different Copper Proteins: A Dynamic Fluorescence Study on Azurin and Ascorbate Oxidase

Tryptophan fluorescence is extremely useful to monitor structural conformational transitions in proteins. Denaturant-induced unfolding of azurin and ascorbate oxidase has been studied by dynamic fluorescence measurements in the frequency domain and the results have been interpreted in terms of continuous distribution of lifetimes. The data add new information on the unfolding mechanism that was previously analyzed by steady-state emission spectroscopy. In particular, the existence of multiple, parallel unfolding pathways may be envisaged and correlated, in both cases, to the two protein structures. The effect of metal depletion has been also characterized by fluorescence lifetime measurements. In the case of azurin, a monomeric protein, the data demonstrate that copper removal yields a totally different unfolding pathways with respect to the holo protein, indicating that metal ion plays a fundamental structural role in the wild type, native protein. In the case of ascorbate oxidase a dimer of 140 kDa, only minor effects have been detected by copper removal. However, the analysis of the fluorescence decay in presence of different amounts of guanidinium hydrochloride gives new important insights on the unfolding intermediates. In particular the data support the hypothesis of a partial exposure of an outer layer of dimer at intermediate denaturant concentration. This ability of dynamic fluorescence to pinpoint the presence of structural micro-heterogeneity in the unfolding pathways of proteins demonstrates the greater power of this technique compared to the most commonly used steady-state measurements.     

Polystyrene Microspheres in Tissue-Simulating Phantoms Can Collisionally Quench Fluorescence

Tissue-simulating phantoms that replicate intrinsic optical properties in a controlled manner are useful for quantitative studies of photon transport in turbid biological media. In such phantoms, polystyrene microspheres are often used to simulate tissue optical scattering. Here, we report that using polystyrene microspheres in fluorescent tissue-simulating phantoms can reduce fluorophore quantum yield via collisional quenching. Fluorescence lifetime spectroscopy was employed to characterize quenching in phantoms consisting of a fluorescein dye and polystyrene microspheres (scattering coefficients _s 100-600cm^–1). For this range of tissue-simulating phantoms, analysis using the Stern-Volmer equation revealed that collisional quenching by polystyrene microspheres accounted for a decrease in fluorescence intensity of 6-17% relative to the intrinsic intensity value when no microspheres (quenchers) were present. The intensity decrease from quenching is independent of additional, anticipated losses arising from optical scattering associated with the microspheres. These results suggest that quantitative fluorescence measurements in studies employing such phantoms may be influenced by collisional quenching.     

Measurement of small spin-spin splittings in the presence of chemical exchange: case of deuteriated water

A new implementation of a described data processing for measuring small scalar coupling constant J is proposed. It is applied particularly to situations where the coupling constant is around the linewidth at half-height, and in the presence of chemical exchange. This modified data processing uses only a simple "one-pulse" experiment instead of a series of spin echoes experiments required by the previous processing. The FID recorded in the one pulse experiment is used by a reconstitution program to generate a set of signals, which are analyzed in the time domain to obtain a spectrum where the scalar coupling constant is apparently multiplied by n+1, where n is positive. The new processing is tested with simulated spectra. The coupling constant between proton and deuterium is measured in the proton spectrum of a solution of 80% of D(2)O and 20% of H(2)O. It was found to be J=1.54+/-0.01 Hz.     

Long-Lived Luminescent Re(I) Complexes Containing cis-Carbonyl and Bidentate Phosphine Ligands

Highly emissive Re(I) complexes, hydrophobic cis-Re(CO)₂(c-dppene)(dpphen) and water-soluble cis-Re(CO)₂(c-dppene)(SO₃-dpphen) with a negative charge, were synthesized [where c-dppene is cis-(bis(diphenylphosphino)ethylene, dpphen is 4,7-diphenyl-1,10-phenanthroline, and SO₃-dpphen is its disulfonate derivative]. These Re(I) complexes have significantly higher molar absorption in the 350- to 490-nm region compared to their tricarbonyl Re(I) counterparts and their emission spectra are similar to those of Ru(II) polypyridyl complexes. The luminescence lifetimes of these Re(I) complexes approach 10 s in the absence of oxygen in both aqueous and less polar solvents. The complexes have limiting anisotropy values exceeding 0.3. As both ligands, the diimine and the bidentate phosphine, can be modified by adding different substituents, it should be possible to make cis-Re(CO)₂(diimine)(P-P) derivatives which are capable of labeling biomacromolecules for biochemical and biophysical studies.