Digital Analysis Of Microscopic Images Research Articles

Application of imaging and spectroscopy techniques for grading of bovine embryos - a review.

Although embryo transfers have grown considerably in the cattle industry, the selection of embryos required for successful pregnancies remains a challenging task. Visual inspection of 7th-day embryos using a stereomicroscope, followed by classification based on morphological features is the most commonly practiced procedure. However, there are inaccuracies and inconsistencies in the manual grading of bovine embryos. The objective of this review was to evaluate the potential of imaging and spectroscopic techniques in the selection of bovine embryos. Digital analysis of microscopic images through extracting visual features in the embryo region, and classification using machine learning methods have yielded about 88-96% success in pregnancies. The Raman spectral pattern provides valuable information regarding developmental stages and quality of the embryo. The Raman spectroscopy approach has also been successfully used to determine various parameters of bovine oocytes. Besides, Fourier Transform Infrared (FTIR) spectroscopy has the ability to assess embryo quality through analyzing embryo composition, including nucleic acid and amides present. Hyperspectral Imaging has also been used to characterize metabolite production during embryo growth. Although the time-lapse imaging approach is beneficial for morphokinetics evaluation of embryo development, optimized protocols are required for successful implementation in bovine embryo transfers. Most imaging and spectroscopic findings are still only at an experimental stage. Further research is warranted to improve the repeatability and practicality to implement in commercial facilities.

Use of ImageJ Software for Assessment of Mechanical Damage to Starch Granules

This study attempted to assess the influence of mechanical forces on potato, tapioca, wheat, rice, and maize starch granules. For this purpose, we used digital analysis of microscopic images of starch granules before and after starch grinding using ImageJ software. Additionally, we studied the influence of temperature on the size and shape of starch granules by drying the starches for 30 min at 60 °C. Our results indicate that mechanical forces very rarely cause damage to starch granules, such as breaking or cracking. In most cases, the action of mechanical forces results only in smoother shape of starch granules and their shrinking, linked with rising temperature. Results of this study show that ImageJ software can be successfully used to assess starch granule size and shape.

Kinetic model to describe the morphological evolution of filamentous fungi during their early stages of growth in the standard submerged and microparticle-enhanced cultivations.

Biosynthesis of metabolites and enzymes by filamentous fungi depends on their morphological form in submerged cultures. However, their early stages of growth lasting approximately 24h, from the introduction of spores to the medium until the formation of stable morphological forms, such as clumps or pellets, have rarely been the objects of experimental and modeling studies. Microparticle-enhanced cultivation (MPEC) has been applied only to a few fungal species, mainly Aspergilli. Therefore, the objective of this work was to formulate the kinetic model to describe the early stages of the fungal evolution in the standard cultivation and MPEC for Aspergillus terreus, Chaetomium globosum, Penicillium rubens, and Mucor racemosus. These fungi exhibit various mechanisms of agglomerates formation in submerged cultures. The experiments were performed in batch shake flasks (parameters identification) and a stirred tank bioreactor (model verification). In the balance equation for fungal cells, the mean projected area of hyphal objects measured by the digital analysis of microscopic images was used as the dependent variable. The analysis of the experimental data and model solution revealed that the effect of the microparticles (aluminum oxide at 6gL-1) in MPEC toward the studied filamentous fungi was to the high extent species dependent. This effect was most evident in the case of spore coagulative A. terreus and noncoagulative M. racemosus.

Morphological evolution of various fungal species in the presence and absence of aluminum oxide microparticles: Comparative and quantitative insights into microparticle-enhanced cultivation (MPEC).

The application of microparticle‐enhanced cultivation (MPEC) is an attractive method to control mycelial morphology, and thus enhance the production of metabolites and enzymes in the submerged cultivations of filamentous fungi. Unfortunately, most literature data deals with the spore‐agglomerating species like aspergilli. Therefore, the detailed quantitative study of the morphological evolution of four different fungal species (Aspergillus terreus, Penicillium rubens, Chaetomium globosum, and Mucor racemosus) based on the digital analysis of microscopic images was presented in this paper. In accordance with the current knowledge, these species exhibit different mechanisms of agglomerates formation. The standard submerged shake flask cultivations (as a reference) and MPEC involving 10 μm aluminum oxide microparticles (6 g·L−1) were performed. The morphological parameters, including mean projected area, elongation, roughness, and morphology number were determined for the mycelial objects within the first 24 hr of growth. It occurred that heretofore observed and widely discussed effect of microparticles on fungi, namely the decrease in pellet size, was not observed for the species whose pellet formation mechanism is different from spore agglomeration. In the MPEC, C. globosum developed core‐shell pellets, and M. racemosus, a nonagglomerative species, formed the relatively larger, compared to standard cultures, pellets with distinct cores.

Hemorheological invitro action of propofol on erythrocytes from healthy donors and diabetic patients.

Drugs used during anesthesia might induce disturbance on microcirculation due to their systemic cardiovascular actions and to direct hemorheological effects. A comparative investigation of the hemorheological alterations related to in vitro propofol treatment of red blood cells (RBCs) from healthy and diabetic volunteers is presented here. Viscoelasticity and aggregation of RBCs from type 2 diabetic patients (DBT) and healthy donors (HD) were studied from RBCs incubated with propofol near steady-state concentration. 'S parameter', which measures the aggregation degree, was obtained using digital analysis of microscopic images. Erythrocyte viscoelasticity parameters were determined using an Erythrocyte Rheometer. Results obtained from DBT samples showed an increase of 10% or more in aggregation due to the propofol action. The phase shift between erythrocyte response and oscillating shear stress applied at 1 Hz was altered by propofol treatment of erythrocyte from HD and DBT. Propofol could produce slight alterations in the rheological behavior of erythrocyte from HD and DBT, at concentrations near those of steady state. Moreover, this anesthetic could induce an adverse effect in DBT, particularly on erythrocyte aggregation. The observed hemorheologic alteration would increase the possibility of microcapillary obstruction. Hence, this type of study [0] would prove relevant to avoid possible postoperative complications.

Application of Image Analysis to Identify Quartz Grains in Heavy Aggregates Susceptible to ASR in Radiation Shielding Concrete.

Alkali-silica reaction (ASR) is considered as a potential aging-related degradation phenomenon that might impair the durability of concrete in nuclear containments. The objective of this paper is the application of digital analysis of microscopic images to identify the content and size of quartz grains in heavy mineral aggregates. The range of investigation covered magnetite and hematite aggregates, known as good absorbers of gamma radiation. Image acquisition was performed using thin sections observed in transmitted cross-polarized light with λ plate. Image processing, consisting of identification of ferrum oxide and epoxy resin, and the subsequent application of a set of filtering operations resulted in an adequate image reduction allowing the grain size analysis. Quartz grains were classified according to their mean diameter so as to identify the reactive range. Accelerated mortar bar tests were performed to evaluate the ASR potential of the aggregates. The SiO2 content in the heavyweight aggregates determined using the image analysis of thin sections was similar to XRF test result. The content of reactive quartz hematite was 2.7%, suggesting that it would be prone to ASR. The expansion test, according to ASTM C1260, confirmed the prediction obtained using the digital image analysis.

Morphologically structured model for growth and citric acid accumulation by Aspergillus niger

A morphologically structured model for the batch process of biomass growth and citric acid accumulation by Aspergillus niger is presented in this paper. The model consists of ten ordinary differential equations, which balance biomass and four physiological zones of hyphae, and includes the most important medium components, such as carbon sources, nitrogen source and citric acid. Digital analysis of microscopic images was employed to determine the hyphal fractions within each physiological zone. All model parameters were determined on the basis of data obtained in four experiments carried out in two types of bioreactors and under various process conditions, specifically normal and increased pressure. Some parameters were directly calculated from the experimental data by means of linear and non-linear regression. This allowed minimisation of the number of parameters to be found by means of an optimisation procedure, down to 11 and thus, significantly reduced computation time. The simulated curves are largely consistent with the experimental data and some parameters appeared to be practically independent of process conditions. These findings contribute to the universality of the model proposed.

Direct gene transfer into rat liver cells by in vivo electroporation

In vivo electro-transfection efficiency and manner of transferred gene expression were investigated by fluorescence microscopic image analysis. Green fluorescent protein (GFP) gene was used as the genetic marker. Electroporation was carried out on the liver of live rats by use of disk electrodes mounted in the tips of tweezers, which were directly pressed onto the surface of a liver lobe in situ. Electroporation with eight electric pulses of 50 ms in duration at 50 V gave a good efficiency of transfection as judged by the induced GFP expression. Bright fluorescence of GFP appeared as dots, which were scattered around the area damaged by electroporation. The transfection efficiency increased as the amount of injected DNA was increased. The results indicate that the amount of induced gene expression can be controlled. Estimation of the efficiency of electro-gene transfer using the fluorescence of GFP and digital analysis of microscopic images was useful to determine the optimum conditions for local gene therapy in tissues and organs.