In-vivo confocal scanning laser microscopy of the cerebral microcirculation

Confocal scanning laser microscopy (CSLM) was used to study the microcirculation of the brain neocortex in anaesthetized rats. After removal of the dura mater, implantation of a closed cranial window, and intravenous injection of fluorescein, three-dimensional reconstructions of cortical capillaries were performed down to a depth of 250 μm below the pial surface. Using a one-dimensional approach (single line scanning), erythrocyte (negative contrast in fluorescently labelled plasma) and leucocyte (labelled with rhodamine 6 G) velocity and supply rate in cortical capillaries were measured. The effect of CO₂-inhalation on capillary blood flow dynamics was studied. Capillaries were imaged continuously for up to 1 h without changes in flow or fluorescence pattern. However, by increasing the laser power 10–100-fold, aggregate formation was induced and capillaries were occluded, possibly due to damage to vascular endothelium. We conclude that CSLM can be used to study morphological and dynamic aspects of fluorescently labelled subsurface structures in organs of experimental animals.     

Microscopy under pressure--an optical chamber system for fluorescence microscopic analysis of living cells under high hydrostatic pressure

High hydrostatic pressure (HHP) becomes more and more interesting for life science research, since it can be employed to inactivate various cells. To directly monitor "cells under pressure," the development of an optical high-pressure chamber is required. Therefore, an optical pressure chamber that can be used for up to 300 MPa was constructed. This chamber has already been described as a tool for in situ observation of dynamic changes of microscopic structures in bright field as well as phase contrast. In combination with an inverted microscope, we obtained brilliant microscopic color pictures with an optical resolution more than 0.56 microm. Here, we demonstrate the capabilities of the HHP cell, in combination with epifluorescence microscopy. Using a nonadherent human B-cell line (Raji, ATCC CCL 86), stained with the fluorescent dyes propidium iodide, Hoechst 33342, or dihexyloxacarbocyanine iodide, we were able to show that the system is suitable to perform fluorescence microscopic analyses, with pressures up to 300 MPa, with viable mammalian cells.     

Real-time contactless measurement of robot pose in six degrees of freedom

For dynamically measuring position in three degrees of freedom laser tracking systems are well known. Up to now no possibility to measure also orientation using the same laser beam has been known. We present a technique to incorporate orientation measurement into a laser tracking system that requires only minor changes in the existing hardware. The method is based on the analysis of an image of the reflected laser beam intensity distribution. In this image the edges of a slightly modified retroreflector represent a function of the orientation of the end-effector holding the retroreflector. It is shown that from this image the orientation can be determined uniquely and in real time.

A laser tracking system including position and orientation measurement constitutes an instrument to accurately determine robot performance as well as to acquire hints on how to improve robot models and control algorithms.     

Swarm-intelligent foraging in honeybees: benefits and costs of task-partitioning and environmental fluctuations

For honeybee colonies, it is crucial to collect nectar in an efficient way. Empiric experiments showed that the process of decision making, which allows the colony to select the optimal nectar source, is based on individual decisions. These decisions are made by returning nectar foragers, which alter their dancing behaviours based on the nectar source’s quality and based on the experienced search time for a receiver bee. Nectar receivers, which represent a shared limited resource for foragers, can modulate the foraging decisions performed by the colony. We investigated the interplay between foragers and receivers by using a multi-agent simulation. Therefore, we implemented agents which are capable of a limited set of behaviours and which spend energy according to their behaviour. In simulation experiments, we tested colonies with various receiver-to-forager ratios and measured colony-level results like the emerging foraging patterns and the colony’s net honey gain. We show that the number of receivers prominently regulates the foraging workforce. All tested environmental fluctuations are predicted to cause energetic costs for the colony. Task-partitioning additionally influences the colony’s decision-making concerning the question whether or not the colony sticks to a nectar source after environmental fluctuations.     

Non-targeted analysis of tea by hydrophilic interaction liquid chromatography and high resolution mass spectrometry

Tea is the second most consumed beverage in the world and its consumption has been associated with numerous potential health benefits. Factors such as fermentation methods, geographical origin and season can affect the primary and secondary metabolite composition of tea. In this study, a hydrophilic interaction liquid chromatography (HILIC) method coupled to high resolution mass spectrometry in both positive and negative ionisation modes was developed and optimised. The method when combined with principal component analysis to analyse three different types of tea, successfully distinguished samples into different categories, and provided evidence of the metabolites which differed between them. The accurate mass and high resolution attributes of the mass spectrometric data were utilised and relative quantification data were extracted post-data acquisition on 18 amino acids, showing significant differences in amino acid concentrations between tea types and countries. This study highlights the potential of HILIC chromatography combined with non-targeted mass spectrometric methods to provide a comprehensive understanding of polar metabolites in plant extracts.     

An NMR-based metabolomic assessment of cultured cobia health in response to dietary manipulation

Commercial aquaculture feeds rely heavily on fishmeal and fish oil, which can be expensive and ecologically unsustainable. To evaluate the efficacy of reduced fishmeal diets for outgrowth, a dietary study was conducted on the finfish cobia, Rachycentron canadum. NMR-based metabolomic techniques were used to assess the effect of decreasing dietary fishmeal on the health of the cobia. Filtered serum ¹H NMR spectra analysed by principal components analysis (PCA) showed cobia fed reduced fishmeal diets were metabolically different than cobia on control diets. In particular, tyrosine and betaine increased in cobia fed reduced fishmeal diets while glucose decreased, suggesting that these cobia were not receiving the necessary nutritional components required for energy and growth. The formulated control diet contributed to enriched growth and significantly elevated lactate levels suggesting enhanced gut microflora metabolism in response to dietary components. The results show that NMR-based metabolomic analysis is a useful tool in aquaculture studies.     

Adaptive controller placement for wireless sensor–actuator networks with erasure channels

Wireless sensor–actuator networks offer flexibility for control design. One novel element which may arise in networks with multiple nodes is that the role of some nodes does not need to be fixed. In particular, there is no need to pre-allocate which nodes assume controller functions and which ones merely relay data. We present a flexible architecture for networked control using multiple nodes connected in series over analog erasure channels without acknowledgments. The control architecture proposed adapts to changes in network conditions, by allowing the role played by individual nodes to depend upon transmission outcomes. We adopt stochastic models for transmission outcomes and characterize the distribution of controller location and the covariance of system states. Simulation results illustrate that the proposed architecture has the potential to give better performance than limiting control calculations to be carried out at a fixed node.