Using flow cytometry for in situ monitoring of antimicrobial compound production in the biocontrol bacterium Pseudomonas fluorescens CHA0.

Pseudomonas fluorescens strain CHA0 is able to protect plants against a variety of pathogens, notably by producing the two antimicrobial compounds 2,4-diacetylphloroglucinol (DAPG) and pyoluteorin (PLT). The regulation of the expression of these compounds is affected by many biotic factors, such as fungal pathogens, rhizosphere bacteria as well as plant species. Therefore, the influence of some plant phenolic compounds on the expression of DAPG and PLT biosynthetic genes has been tested using GFP-based reporter, monitored by standard fluometry and flow cytometry. In situ experiments were also performed with cucumber plants. We found that several plant metabolites such as IAA and umbelliferone are able to modify significantly the expression of DAPG and PLT. The use of flow cytometry with autofluorescents proteins seems to be a promising method to study rhizobacteria-plant interactions.

Artificial neural network study of whole-cell bacterial bioreporter response determined using fluorescence flow cytometry.

Genetically engineered bioreporters are an excellent complement to traditional methods of chemical analysis. The application of fluorescence flow cytometry to detection of bioreporter response enables rapid and efficient characterization of bacterial bioreporter population response on a single-cell basis. In the present study, intrapopulation response variability was used to obtain higher analytical sensitivity and precision. We have analyzed flow cytometric data for an arsenic-sensitive bacterial bioreporter using an artificial neural network-based adaptive clustering approach (a single-layer perceptron model). Results for this approach are far superior to other methods that we have applied to this fluorescent bioreporter (e.g., the arsenic detection limit is 0.01 microM, substantially lower than for other detection methods/algorithms). The approach is highly efficient computationally and can be implemented on a real-time basis, thus having potential for future development of high-throughput screening applications.

Colour image processing

In the context of the round table the following topics related to image colour processing will be discussed: historical point of view. Studies of Aguilonius, Gerritsen, Newton and Maxwell. CIE standard (Commission International de lpsilaEclaraige). Colour models. RGB, HIS, etc. Colour segmentation based on HSI model. Industrial applications. Summary and discussion. At the end, video images showing the robustness of colour in front of B/W images will be presented

A method to obtain the integrated contour image in colour scenes

This paper describes a method to achieve the most relevant contours of an image. The presented method proposes to integrate the information of the local contours from chromatic components such as H, S and I, taking into account the criteria of coherence of the local contour orientation values obtained from each of these components. The process is based on parametrizing pixel by pixel the local contours (magnitude and orientation values) from the H, S and I images. This process is carried out individually for each chromatic component. If the criterion of dispersion of the obtained orientation values is high, this chromatic component will lose relevance. A final processing integrates the extracted contours of the three chromatic components, generating the so-called integrated contours image

An approach to vision-based station keeping for an unmanned underwater vehicle

This paper presents an automatic vision-based system for UUV station keeping. The vehicle is equipped with a down-looking camera, which provides images of the sea-floor. The station keeping system is based on a feature-based motion detection algorithm, which exploits standard correlation and explicit textural analysis to solve the correspondence problem. A visual map of the area surveyed by the vehicle is constructed to increase the flexibility of the system, allowing the vehicle to position itself when it has lost the reference image. The testing platform is the URIS underwater vehicle. Experimental results demonstrating the behavior of the system on a real environment are presented

Towards Detecting Changes in Underwater Image Sequences

Detecting changes between images of the same scene taken at different times is of great interest for monitoring and understanding the environment. It is widely used for on-land application but suffers from different constraints. Unfortunately, Change detection algorithms require highly accurate geometric and photometric registration. This requirement has precluded their use in underwater imagery in the past. In this paper, the change detection techniques available nowadays for on-land application were analyzed and a method to automatically detect the changes in sequences of underwater images is proposed. Target application scenarios are habitat restoration sites, or area monitoring after sudden impacts from hurricanes or ship groundings. The method is based on the creation of a 3D terrain model from one image sequence over an area of interest. This model allows for synthesizing textured views that correspond to the same viewpoints of a second image sequence. The generated views are photometrically matched and corrected against the corresponding frames from the second sequence. Standard change detection techniques are then applied to find areas of difference. Additionally, the paper shows that it is possible to detect false positives, resulting from non-rigid objects, by applying the same change detection method to the first sequence exclusively. The developed method was able to correctly find the changes between two challenging sequences of images from a coral reef taken one year apart and acquired with two different cameras

Estimating the motion of an underwater robot from a monocular image sequence

When underwater vehicles perform navigation close to the ocean floor, computer vision techniques can be applied to obtain quite accurate motion estimates. The most crucial step in the vision-based estimation of the vehicle motion consists on detecting matchings between image pairs. Here we propose the extensive use of texture analysis as a tool to ameliorate the correspondence problem in underwater images. Once a robust set of correspondences has been found, the three-dimensional motion of the vehicle can be computed with respect to the bed of the sea. Finally, motion estimates allow the construction of a map that could aid to the navigation of the robot

Correction of shading effects in vision-based UUV localization

This paper describes the improvements achieved in our mosaicking system to assist unmanned underwater vehicle navigation. A major advance has been attained in the processing of images of the ocean floor when light absorption effects are evident. Due to the absorption of natural light, underwater vehicles often require artificial light sources attached to them to provide the adequate illumination for processing underwater images. Unfortunately, these flashlights tend to illuminate the scene in a nonuniform fashion. In this paper a technique to correct non-uniform lighting is proposed. The acquired frames are compensated through a point-by-point division of the image by an estimation of the illumination field. Then, the gray-levels of the obtained image remapped to enhance image contrast. Experiments with real images are presented

Towards a real-time vision-based navigation system for a small-class UUV

This paper deals with the problem of navigation for an unmanned underwater vehicle (UUV) through image mosaicking. It represents a first step towards a real-time vision-based navigation system for a small-class low-cost UUV. We propose a navigation system composed by: (i) an image mosaicking module which provides velocity estimates; and (ii) an extended Kalman filter based on the hydrodynamic equation of motion, previously identified for this particular UUV. The obtained system is able to estimate the position and velocity of the robot. Moreover, it is able to deal with visual occlusions that usually appear when the sea bottom does not have enough visual features to solve the correspondence problem in a certain area of the trajectory

Monocular-based 3-D seafloor reconstruction and ortho-mosaicing by piecewise planar representation

Photo-mosaicing techniques have become popular for seafloor mapping in various marine science applications. However, the common methods cannot accurately map regions with high relief and topographical variations. Ortho-mosaicing borrowed from photogrammetry is an alternative technique that enables taking into account the 3-D shape of the terrain. A serious bottleneck is the volume of elevation information that needs to be estimated from the video data, fused, and processed for the generation of a composite ortho-photo that covers a relatively large seafloor area. We present a framework that combines the advantages of dense depth-map and 3-D feature estimation techniques based on visual motion cues. The main goal is to identify and reconstruct certain key terrain feature points that adequately represent the surface with minimal complexity in the form of piecewise planar patches. The proposed implementation utilizes local depth maps for feature selection, while tracking over several views enables 3-D reconstruction by bundle adjustment. Experimental results with synthetic and real data validate the effectiveness of the proposed approach

Behaviours implemented using a vision system based on coded light projection

In a search for new sensor systems and new methods for underwater vehicle positioning based on visual observation, this paper presents a computer vision system based on coded light projection. 3D information is taken from an underwater scene. This information is used to test obstacle avoidance behaviour. In addition, the main ideas for achieving stabilisation of the vehicle in front of an object are presented

Shape complexity based on mutual information

Shape complexity has recently received attention from different fields, such as computer vision and psychology. In this paper, integral geometry and information theory tools are applied to quantify the shape complexity from two different perspectives: from the inside of the object, we evaluate its degree of structure or correlation between its surfaces (inner complexity), and from the outside, we compute its degree of interaction with the circumscribing sphere (outer complexity). Our shape complexity measures are based on the following two facts: uniformly distributed global lines crossing an object define a continuous information channel and the continuous mutual information of this channel is independent of the object discretisation and invariant to translations, rotations, and changes of scale. The measures introduced in this paper can be potentially used as shape descriptors for object recognition, image retrieval, object localisation, tumour analysis, and protein docking, among others

An information theoretic framework for image segmentation

In this paper, an information theoretic framework for image segmentation is presented. This approach is based on the information channel that goes from the image intensity histogram to the regions of the partitioned image. It allows us to define a new family of segmentation methods which maximize the mutual information of the channel. Firstly, a greedy top-down algorithm which partitions an image into homogeneous regions is introduced. Secondly, a histogram quantization algorithm which clusters color bins in a greedy bottom-up way is defined. Finally, the resulting regions in the partitioning algorithm can optionally be merged using the quantized histogram

Estimation of the lateral correlation structure of subsurface water content from surface-based ground-penetrating radar reflection images

Over the past decade, significant interest has been expressed in relating the spatial statistics of surface-based reflection ground-penetrating radar (GPR) data to those of the imaged subsurface volume. A primary motivation for this work is that changes in the radar wave velocity, which largely control the character of the observed data, are expected to be related to corresponding changes in subsurface water content. Although previous work has indeed indicated that the spatial statistics of GPR images are linked to those of the water content distribution of the probed region, a viable method for quantitatively analyzing the GPR data and solving the corresponding inverse problem has not yet been presented. Here we address this issue by first deriving a relationship between the 2-D autocorrelation of a water content distribution and that of the corresponding GPR reflection image. We then show how a Bayesian inversion strategy based on Markov chain Monte Carlo sampling can be used to estimate the posterior distribution of subsurface correlation model parameters that are consistent with the GPR data. Our results indicate that if the underlying assumptions are valid and we possess adequate prior knowledge regarding the water content distribution, in particular its vertical variability, this methodology allows not only for the reliable recovery of lateral correlation model parameters but also for estimates of parameter uncertainties. In the case where prior knowledge regarding the vertical variability of water content is not available, the results show that the methodology still reliably recovers the aspect ratio of the heterogeneity.

Methods for the ex vivo characterization of human CD8+ T subsets based on gene expression and replicative history analysis.

The generation of an antigen-specific T-lymphocyte response is a complex multi-step process. Upon T-cell receptor-mediated recognition of antigen presented by activated dendritic cells, naive T-lymphocytes enter a program of proliferation and differentiation, during the course of which they acquire effector functions and may ultimately become memory T-cells. A major goal of modern immunology is to precisely identify and characterize effector and memory T-cell subpopulations that may be most efficient in disease protection. Sensitive methods are required to address these questions in exceedingly low numbers of antigen-specific lymphocytes recovered from clinical samples, and not manipulated in vitro. We have developed new techniques to dissect immune responses against viral or tumor antigens. These allow the isolation of various subsets of antigen-specific T-cells (with major histocompatibility complex [MHC]-peptide multimers and five-color FACS sorting) and the monitoring of gene expression in individual cells (by five-cell reverse transcription-polymerase chain reaction [RT-PCR]). We can also follow their proliferative life history by flow-fluorescence in situ hybridization (FISH) analysis of average telomere length. Recently, using these tools, we have identified subpopulations of CD8+ T-lymphocytes with distinct proliferative history and partial effector-like properties. Our data suggest that these subsets descend from recently activated T-cells and are committed to become differentiated effector T-lymphocytes.