Luminescent lanthanide complexes display unrivalled spectroscopic properties, which place them in a special category in the luminescent toolbox. Their long-lived line-like emission spectra are the cornerstones of numerous analytical applications ranging from ultrasensitive homogeneous fluoroimmunoassays to the study of molecular interactions in living cells with multiplexed microscopy. However, achieving such minor miracles is a result of years of synthetic efforts and spectroscopic studies to understand and gather all the necessary requirements for the labels to be efficient. This feature article intends to survey these criteria and to discuss some of the most important examples reported in the literature, before explaining in detail some of the applications of luminescent lanthanide labels to bioanalysis and luminescence microscopy. Finally, the emphasis will be put on some recent applications that hold great potential for future biosensing.

Piling up excited states to reach upconversion (UC) is severely restricted by vibrational quenching mechanisms, especially when one looks at discrete molecular entities in solution. By carefully controlling the supramolecular assembly processes resulting from the strong electrostatic interactions between negatively charged Yb complexes and Tb 3+ cations in aqueous solutions, we engineered the formation of heteropolynuclear complexes of [(YbL) 2 Tb x ] compositions (x = 1 and 2). These edifices display a phenomenon of cooperative photosensitization UC with green emission of the Tb cations upon NIR excitation at 980 nm in the Yb absorption band. The photophysical properties of the complexes were carefully investigated by steady-state and time-resolved luminescence experiments in D 2 O, allowing to quantify the impact of the composition and pD of the solution on the emission intensity, as well as clarifying the exact cooperative photosensitization upconversion mechanism. Using optimized conditions, the energy transfer UC process could be observed for the first time in non-deuterated water with discrete molecular compounds.

The new potentially octadentate ligand, 1-(carboxymethyl)-4,7-bis[(6-carboxypyridin-2-yl)methyl]-1,4,7-triazacyclononane (H(3)bpatcn), in which two picolinate arms and one acetate arm are connected to the 1,4,7-triazacyclonane core, has been prepared. Potentiometric studies show an increased stability of the Gd(III) complex of H(3)bpatcn (logK(GdL)=15.8(2)) with respect to the Gd(III) complex of the analogous ligand 1,4,7-triazacyclononane-N,N',N''-triacetic acid (H(3)nota) (logK(GdL)=13.7), associated with an increased selectivity of H(3)bpatcn for gadolinium over calcium. The H(3)bpatcn ligand sensitises the terbium ion very efficiently, leading to a long-lived and highly luminescent terbium complex (quantum yield=43 %), in spite of the presence of a coordinated water molecule. (1)H proton NMR studies indicate that the metal ion is rigidly encapsulated by the three arms of the octadentate ligand H(3)bpatcn and that the macrocycle framework remains bound (through the five nitrogen and the three oxygen atoms) even at high temperature. A new theoretical method for interpreting the water proton relaxivity is presented. It is based on recent progresses in the description of the electronic spin relaxation and on an auxiliary probe solute. It replaces the Solomon, Bloembergen and Morgan (SBM) framework, which is questionable at low field, while avoiding resorting to simulations and/or sophisticated theories with additional unknown zero-field splitting (ZFS) parameters. The inclusion of two picolinate groups on a triazacyclononane framework affords the mono-aquo gadolinium complex [Gd(bpatcn)(H(2)O)] with favourable electron-relaxation properties (tau(eff)(S0)=125 ps). The optimisation of the electronic relaxation by ligand design is especially important to achieve high relaxivity in the new generation macromolecular complexes with long rotational correlation times.

A new family of mixed-lanthanide (Yb(III) and Nd(III)) transition-metal (f-d) cyclen-Ru(II)(phen)(3) (phen = 1,10-phenanthroline) complexes were synthesized as dual visible- and near-infrared (NIR)-emitting DNA probes/sensors. Significant changes were seen in both the Ru(II) visible and the Yb(III)-centered NIR emission, which was switched off upon binding to DNA at pH 7.4. In contrast, no changes were seen in the Nd(III) emission of the analogue f-d conjugate.

The search for more biocompatible alternatives to Gd 3+-based MRI agents,a nd the interest in 52 Mn for PET imaging call for ligands that form inert Mn 2+ chelates.G iven the labile nature of Mn 2+ ,h igh inertness is challenging to achieve.The strongly preorganized structure of the 2,4-pyridyldisubstituted bispidol ligand L 1 endows its Mn 2+ complex with exceptional kinetic inertness.I ndeed, MnL 1 did not showa ny dissociation for 140 days in the presence of 50 equiv.o fZ n 2+ (37 8 8C, pH 6), while recently reported potential MRI agents MnPyC3A and MnPC2A-EA have dissociation half-lives of 0.285 ha nd 54.4 hu nder similar conditions.I na ddition, the relaxivity of MnL 1 (4.28 mm À1 s À1 at 25 8 8C, 20 MHz) is remarkable for am onohydrated, small Mn 2+ chelate.I nvivo MRI experiments in mice and determination of the tissue Mn content evidence rapid renal clearance of MnL 1 .Additionally, L 1 could be radiolabeled with 52 Mn and the complex revealed good stability in biological media.

Up conversion is an Anti-Stokes luminescent process by which photons of low energy are piled up to generate light at a higher energy. Here we show that the addition of fluoride anions to a D2O solution of a macrocyclic erbium complex leads to the formation of a supramolecular [(ErL)2F]+ assembly in which fluoride is sandwiched between two complexes, held together by the synergistic interactions of the Er-F-Er bridging bond, four intercomplex hydrogen bonds and two aromatic stacking interactions. Room temperature excitation into the Er absorption bands at 980 nm of a solution of the complex in D2O results in the observation of up converted emission at 525, 550 and 650 nm attributed to Er centred transitions via a two-step excitation. The up conversion signal is dramatically increased upon formation of the [(ErL)2F]+ dimer in the presence of 0.5 equivalents of fluoride anions.

Addition of Tb salts to a solution of a (YbL) complex in DO resulted in the formation of [(YbL)Tb] (x = 1 to 3) complexes that, upon NIR excitation at 980 nm, showed an unprecedented Yb to Tb upconversion sensitization phenomenon resulting in the observation of the typical green emission of Tb.

Dedicated to Professor Jean-Claude Bünzli on the occasion of his 65th birthday A series of europium(III) and terbium(III) complexes of three 1,4,7-triazacyclononane-based pyridine containing ligands were synthesized. The three ligands differ from each other in the substitution of the pyridine pendant arm, namely they have a carboxylic acid, an ethylamide, or an ethyl ester substituent, i.e., these ligands are 6,6',6''-[1,4,7-triazacyclononane-1,4,7-triyltris(methylene)]tris[pyridine-2-carboxylic acid] (H 3 tpatcn), -tris[pyridine-2-carboxamide] (tpatcnam), and -tris[pyridine-2-carboxylic acid] triethyl ester (tpatcnes) respectively. The quantum yields of both the europium(III) and terbium(III) emission, upon ligand excitation, were highly dependent upon ligand substitution, with a ca. 50-fold decrease for the carboxamide derivative in comparison to the picolinic acid (¼ pyridine-2-carboxylic acid) based ligand. Detailed analysis of the radiative rate constants and the energy of the triplet states for the three ligand systems revealed a less efficient energy transfer for the carboxamidebased systems. . Cationic lanthanide complexes have also been proposed as chemical-exchange saturation-transfer (CEST) agents for MRI [10 -12]. One of the most important requirements, common to all these applications, is the thermodynamic and kinetic stability of the lanthanide complexes in biological media necessary to prevent the release of free lanthanide ions. Because lanthanide ions show high kinetic lability and form preferentially electrostatic bonds with negatively charged O-or N-donors, highly pre-organized polyaminocarboxylate ligands have been used to