Thresholding Approach for Automatic Detection of Pseudomonas aeruginosa Biofilms from Fluorescence in situ Hybridization Images

Pseudomonas aeruginosa is an opportunistic pathogen that forms surface-associated microbial communities (biofilms) on artificial implant devices and on human tissue. Biofilm infections are difficult to treat with antibiotics, in part, because the bacteria in biofilms are physiologically heterogeneous. One measure of biological heterogeneity in a population of cells is to quantify the cellular concentrations of ribosomes, which can be probed with fluorescently labeled nucleic acids. The fluorescent signal intensity following fluorescence in situ hybridization (FISH) analysis correlates to the cellular level of ribosomes. The goals here are to provide computationally and statistically robust approaches to automatically quantify cellular heterogeneity in biofilms from a large library of epifluorescent microscopy FISH images. In this work, the initial steps were developed toward these goals by developing an automated biofilm detection approach for use with FISH images. The approach allows rapid identification of biofilm regions from FISH images that are counterstained with fluorescent dyes. This methodology provides advances over other computational methods, allowing subtraction of spurious signals and non-biological fluorescent substrata. This method will be a robust and user-friendly approach which will enable users to semi-automatically detect biofilm boundaries and extract intensity values from fluorescent images for quantitative analysis of biofilm heterogeneity.

Numerical Simulation of Bio-Chemical Diffusion in Bone Scaffolds

Previously, some materials like solid metals and their alloys have been used as implants in human’s body. In order to amend fixation of these artificial hard human tissues, some porous structures have been introduced. In this way, tissues in vicinity of the porous structure can be attached more easily to the inserted implant. In particular, the porous bone scaffolds are useful since they can deliver important biomolecules like growth factors and proteins. This study focuses on the properties of the degradable porous hard tissues using a three-dimensional numerical Finite Element Method (FEM). The most important studied properties of these structures are diffusivity flux and concentration of different species like glucose, oxygen, and lactate. The process of cells migration into the scaffold is considered as a diffusion process, and related parameters are studied for different values of production/consumption rates.

Low Cost Technique for Measuring Luminance in Biological Systems

In this work, the relationship between the melanin content in a tissue and subsequent absorption of light through that tissue was determined using a digital camera. This technique proved to be simple, cost effective, efficient and reliable. Tissue phantom samples were created using milk and soy sauce to simulate the optical properties of melanin content in human tissue. Increasing the concentration of soy sauce in the milk correlated to an increase in melanin content of an individual. Two methods were employed to measure the light transmitted through the sample. The first was direct measurement of the transmitted intensity using a conventional lux meter. The second method involved correctly calibrating an ordinary digital camera and using image analysis software to calculate the transmitted intensity through the phantom. The results from these methods were then graphically compared to the theoretical relationship between the intensity of transmitted light and the concentration of absorbers in the sample. Conclusions were then drawn about the effectiveness and efficiency of these low cost methods.

Analysis of the Internal Mechanical Conditions in the Lower Limb Due to External Loads

Human soft tissue is loaded and deformed by any activity, an effect known as a stress-strain relationship, and is often described by a load and tissue elongation curve. Several advances have been made in the fields of biology and mechanics of soft human tissue. However, there is limited information available on in vivo tissue mechanical characteristics and behavior. Confident mechanical properties of human soft tissue cannot be extrapolated from e.g. animal testing. Thus, there is need for non invasive methods to analyze mechanical characteristics of soft human tissue. In the present study, the internal mechanical conditions of the lower limb, which is subject to an external load, is studied by use of the finite element method. A detailed finite element model of the lower limb is made possible by use of MRI scans. Skin, fat, bones, fascia and muscles are represented separately and the material properties for them are obtained from literature. Previous studies have been shown to address macroscopic deformation features, e.g. indentation depth, to a large extent. However, the detail in which the internal anatomical features have been modeled does not reveal the critical internal strains that may induce hypoxia and/or eventual tissue damage. The results of the present study reveals that lumped material models, i.e. averaging of the material properties for the different constituents, does not capture regions of critical strains in contrast to more detailed models.

Performance Analysis of Reconstruction Algorithms in Diffuse Optical Tomography

Diffuse Optical Tomography (DOT) is a non-invasive imaging modality used in clinical diagnosis for earlier detection of carcinoma cells in brain tissue. It is a form of optical tomography which produces gives the reconstructed image of a human soft tissue with by using near-infra-red light. It comprises of two steps called forward model and inverse model. The forward model provides the light propagation in a biological medium. The inverse model uses the scattered light to collect the optical parameters of human tissue. DOT suffers from severe ill-posedness due to its incomplete measurement data. So the accurate analysis of this modality is very complicated. To overcome this problem, optical properties of the soft tissue such as absorption coefficient, scattering coefficient, optical flux are processed by the standard regularization technique called Levenberg - Marquardt regularization. The reconstruction algorithms such as Split Bregman and Gradient projection for sparse reconstruction (GPSR) methods are used to reconstruct the image of a human soft tissue for tumour detection. Among these algorithms, Split Bregman method provides better performance than GPSR algorithm. The parameters such as signal to noise ratio (SNR), contrast to noise ratio (CNR), relative error (RE) and CPU time for reconstructing images are analyzed to get a better performance.

Influence of Internal Topologies on Components Produced by Selective Laser Melting: Numerical Analysis

Regardless of the manufacturing process used, subtractive or additive, material, purpose and application, produced components are conventionally solid mass with more or less complex shape depending on the production technology selected. Aspects such as reducing the weight of components, associated with the low volume of material required and the almost non-existent material waste, speed and flexibility of production and, primarily, a high mechanical strength combined with high structural performance, are competitive advantages in any industrial sector, from automotive, molds, aviation, aerospace, construction, pharmaceuticals, medicine and more recently in human tissue engineering. Such features, properties and functionalities are attained in metal components produced using the additive technique of Rapid Prototyping from metal powders commonly known as Selective Laser Melting (SLM), with optimized internal topologies and varying densities. In order to produce components with high strength and high structural and functional performance, regardless of the type of application, three different internal topologies were developed and analyzed using numerical computational tools. The developed topologies were numerically submitted to mechanical compression and four point bending testing. Finite Element Analysis results demonstrate how different internal topologies can contribute to improve mechanical properties, even with a high degree of porosity relatively to fully dense components. Results are very promising not only from the point of view of mechanical resistance, but especially through the achievement of considerable variation in density without loss of structural and functional high performance.

Determination of Electromagnetic Properties of Human Tissues

In this paper a computer system for electromagnetic properties measurements is designed. The system employs Agilent 4294A precision impedance analyzer to measure the amplitude and the phase of a signal applied over a tested biological tissue sample. Measured by the developed computer system data could be used for tissue characterization in wide frequency range from 40Hz to 110MHz. The computer system can interface with output devices acquiring flexible testing process.

Electromagnetic Field Modeling in Human Tissue

For investigations of electromagnetic field distributions in biological structures by Finite Element Method (FEM), a method for automatic 3D model building of human anatomical objects is developed. Models are made by meshed structures and specific electromagnetic material properties for each tissue type. Mesh is built according to specific FEM criteria for achieving good solution accuracy. Several FEM models of anatomical objects are built. Formulation using magnetic vector potential and scalar electric potential (A-V, A) is used for modeling of electromagnetic fields in human tissue objects. The developed models are suitable for investigations of electromagnetic field distributions in human tissues exposed in external fields during magnetic stimulation, defibrillation, impedance tomography etc.