Ruthenium dipyridophenazine (dppz) complexes are sensitive luminescent probes for hydrophobic environments. Here, we apply multiple-frequency fluorescence lifetime imaging microscopy (FLIM) to Δ and Λ enantiomers of lipophilic ruthenium dppz complexes in live and fixed cells, and their different lifetime staining patterns are related to conventional intensity-based microscopy. Excited state lifetimes of the enantiomers determined from FLIM measurements correspond well with spectroscopically measured emission decay curves in pure microenvironments of DNA, phospholipid membrane or a model protein. We show that FLIM can be applied to monitor the long-lived excited states of ruthenium complex enantiomers and, combined with confocal microscopy, give new insight into their biomolecular binding and reveal differences in the microenvironment probed by the complexes.

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International audienceParticle and solute transport by faunal activities may significantly influence rates and pathways of organic matter mineralization during early diagenesis in surface sediments. One of the most frequently utilized techniques to quantify benthic biological reworking activities involves the calculation of a biodiffusion coefficient (Db) estimated from model predictions of 1-D tracer distribution patterns. This technique is labor-intensive and time-demanding. Furthermore, it is normally used for measurements over several days and averages overall transport mechanisms from 3-D to 1-D on a cm scale. In the frame of this work, we developed a new technique based on the nondestructive screening of fluorescent particles (luminophores) using optical discrimination and CCD camera detection of fluorescence (2-D). At a site characterized by a dense population of the brittle star Amphiura filiformis and a high biodiffusion coefficient (obtained from 1-D distributions; Db=35.5±3.7 cm−2 year−1; n=3), the optical reworking coefficient (ORC), estimated from the 2-D luminophore distribution patterns, was calculated (ORC=27.4±9.1 View the MathML source cm−2 h−1; n=24). A nondestructive 2-D approach to quantify particle reworking may provide a powerful and complementary tool to further understand particle transport by the benthic fauna in surface sediments. The optical technique for 2-D detection of luminophores is relatively fast and easy to perform, with the ability to detect small scale (mm) particle movements on a time resolution of minutes or less

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The physical effect of nitrogen upon plants has been studied thoroughly; however, direct studies of nitrogen turnover close to roots have been limited by analytical techniques with low spatial and temporal resolution. Thus, little is known about differences in turnover taking place along and between intact root structures over time as well as how root arrangement, root cell type, plant age, microbial activity, and the dark/light cycle influence uptake and supply of nutrients to root structures. In this study an imaging (planar) optode was used to quantify ammonium over time close to an intact root system of a large fruit bearing tomato plant (Lycopersicon esculentum). Images throughout the experiment made it possible to define the ammonium depletion zone and active turnover potential as well as determine turnover rate and flow patterns around the root system over time. The results indicated that ammonium uptake for tomato plants proceeds over the entire root structure but transverse thin peripheral roots are about twice as efficient as the main root and that the uptake process might influence nutrient availability. The flow patterns close to the root structure revealed that apical regions seem to have a central role in ammonium acquisition.