Reusable Chamber Research Articles

LensHooke® X1PRO comparison with laboratory based manual semen analyses in a Chilean men’s population.

For over 30 years, CASA-Mot technology has been used for kinematic analysis of sperm motility in different mammalian species, but insufficient attention has been paid to the technical limitations of commercial computer-aided sperm analysis (CASA) systems. Counting chamber type and frame rate are two of the most important aspects to be taken into account. Counting chambers can be disposable or reusable, with different depths. In human semen analysis, reusable chambers with a depth of 10 µm are the most frequently used, whereas for most farm animal species it is more common to use disposable chambers with a depth of 20 µm . The frame rate was previously limited by the hardware, although changes in the number of images collected could lead to significant variations in some kinematic parameters, mainly in curvilinear velocity (VCL). A frame rate of 60 frames s−1 is widely considered to be the minimum necessary for satisfactory results. However, the frame rate is species specific and mus...

Imaging Chambers for Arabidopsis Seedlings for Mitotic Studies.

Flowering plants evolved away from creating centrosomes or conventional microtubule organizing centers. Therein, plants have posed a long-standing challenge to many of the conventional ideas for mitotic spindle construction and the process of chromosome segregation. The Arabidopsis seedling has emerged as a leading model for plant cell biological studies of the cytoskeleton and vesicle trafficking. Here we describe methods for creating a reusable chamber for mitotic studies in both seedling root and shoot cells with instruction for best practices with conventional microscopic techniques.

Influence of Particle Size and Particle Range Distribution on the Microstructure of Al–Mg–Si/PKSA Composite for Amateur Solid Rocket Chamber

The study discusses an overview of small rocket program, development and characterization of locally sourced material at low cost. The aim is to enable the construction of small sounding rocket components for experimental purposes in Nigeria. This paper proposes the utilization of palm kernel shell ash as filler materials in the development, characterization and production of composite material to construct prototype reusable chamber and accessories that will enable it possible for small scientific experiments. To achieve this, particle size and particle range distribution on the microstructure, texture, and mechanical properties of (Al–Mg–Si)/PKSA composites developed by powder metallurgy method were investigated. The access in the 6xxx series of aluminum-magnesium-silicon alloy was investigated for this reason. ImageJ software was used to do particle size analysis professionally and the software was used to calculate the area, mean, standard deviation (SD) and the pixel values. The particle size distribution of big constituent (densification or rather densified solids) particles and small dispersoids possess a finer and slightly elongated grain structure when compared with the unreinforced alloy. The results of the XRD and XRF of all the samples considered showed that Al2O3, SiO2, Mn2O3 and MgO phases were common to all. These hard phases are considered to be responsible for improved mechanical properties and resistance of the composite, while the SEM result showed that the reinforcement was uniformly distributed which further improves the mechanical property of the composite.Keywords: Microstructure, Composite, Rocket, Palm Kernel

Standardization of sperm motility analysis by using CASA-Mot for Atlantic salmon (Salmo salar), European eel (Anguilla anguilla) and Siberian sturgeon (Acipenser baerii)

It is essential to define an optimized standard method to assess the fish sperm quality to minimize the differences between the results obtained by different laboratories. Only this optimization and standardization can make them useful from academia to industry. This study presents the validation of sperm motility assessment using a CASA-Mot system for three endangered diadromous fish species: European eel (Anguilla anguilla), Atlantic salmon (Salmo salar) and Siberian sturgeon (Acipenser baerii). To attain this goal, different technical and data processing methods were tested: 1) magnification lens (×10 and ×20), 2) Spermtrack® reusable chambers (10 and 20 μm depth) and 3) different frame rates (50 ≥ FR ≤ 250). The results suggested that the sperm motility assessment for eel, salmon and sturgeon should be performed at 200, 250 and 225 frames s−1, respectively. Moreover, to obtain a high number of analysed spermatozoa in less time and a natural movement of the sperm cells, it is recommended to use ×10 objective and 20 μm depth. In conclusion, different technical settings influence sperm kinetic parameters and should be validated for each fish species to allow the comparison of results between laboratories.

Low-cost equipment for electroformation of Giant Unilamellar Vesicles

<h2>Abstract</h2> Giant Unilamellar Vesicles (GUVs) constitute a very useful model for biomembranes. They are usually generated by electroformation using sinusoidal potential waves applied between two platinum electrodes or electrodes composed of glasses covered by indium tin oxide (ITO). Here we present the design of a programmable, compact and low cost wave generator that allows applying different waveforms, even with initial non-periodical shapes. Thus, it allows to optimize the potential waveform for the particular GUV' composition. We also describe the design of a low cost, reusable chamber with stainless steel electrodes that allows using very small volumes of solution (0.3 mL). With this chamber, a larger number of GUVs were generated in less electroformation times compared to ITO electrodes. The general characteristics of the obtained GUVs are comparable to those electroformed in platinum or ITO surfaces. The chamber, combined with the wave generator comprises a simple, reusable, programmable and low cost system for the formation of GUVs.

CASA-Mot technology: how results are affected by the frame rate and counting chamber.

For over 30 years, CASA-Mot technology has been used for kinematic analysis of sperm motility in different mammalian species, but insufficient attention has been paid to the technical limitations of commercial computer-aided sperm analysis (CASA) systems. Counting chamber type and frame rate are two of the most important aspects to be taken into account. Counting chambers can be disposable or reusable, with different depths. In human semen analysis, reusable chambers with a depth of 10µm are the most frequently used, whereas for most farm animal species it is more common to use disposable chambers with a depth of 20µm . The frame rate was previously limited by the hardware, although changes in the number of images collected could lead to significant variations in some kinematic parameters, mainly in curvilinear velocity (VCL). A frame rate of 60 frames s-1 is widely considered to be the minimum necessary for satisfactory results. However, the frame rate is species specific and must be defined in each experimental condition. In conclusion, we show that the optimal combination of frame rate and counting chamber type and depth should be defined for each species and experimental condition in order to obtain reliable results.

Infrared Techniques for Practical Defect Identification in Bonded Joints in Liquefied Natural Gas Carriers

A robust and reliable method for the identification of defects in adhesively bonded joints used in the membrane containment system used to store liquefied natural gas during transport on board ships is required. The adhesively bonded interface of the membrane system may contain low volume defects known as kissing defects, which are extremely difficult to detect using portable inspection techniques. A novel methodology for detecting such defects is described in the paper. To demonstrate the approach simulated kissing defects were produced in a controlled fashion in a representative secondary membrane bond using silicon grease contamination. The defects were detected using an active thermographic approach known as pulsed phase thermography (PPT) which detects subsurface defects by monitoring the effect they have on the propagation of heat through a component. Due to the low volume of kissing defects, they generally have minimal effect on the heat propagation, so the detection was made possible by application of a small load generated by application of a vacuum. The vacuum can be set-up using a reusable chamber and a standard vacuum pump and therefore is portable and can be applied on-site and in a shipyard during construction.

Efecto de la cámara de recuento utilizada sobre los parámetros espermáticos, analizados con el ISASv1®

IntroductionThe use of counting chambers conditions the sperm concentration and motility values by their type of filling (capillary or drop displacement) and chamber height. We have recently introduced new chambers, both disposable (ISAS®D4C) and reusable (Spermtrack®), with different heights in each case. ObjectiveThis study has aimed to validate its use by analyzing different sperm parameters with a CASA system, that is, the ISASv1®. Material and methodSamples from volunteer donors (10 for the disposable and 5 for reusable), analyzing the effect of chamber type, height, filling and analysis time, were used. ResultsThe results obtained showed no differences within the filling time for the reusable chambers, or the analysis time in any cases. DiscussionThe 10 microns reusable chamber and disposable, 10, 16 and 20 microns showed no differences between them. However, the reusable 20 microns showed significantly different results, so its use (as recommended by WHO) should be taking this into account. ConclusionThe new chambers offer very consistent results, independent of filling or time of analysis.

The performance of 10 different methods for the estimation of sperm concentration

Objective: To evaluate the performance of different methods of sperm counting using latex beads and sperm suspension. Design: A quality-control study. Setting: University-based andrology laboratory. Intervention(s): None. Main Outcome Measure(s): Counting latex beads six times in two standard suspensions using 10 methods and counting spermatozoa with 4 methods. Result(s): When air-displacement pipettes were used, the disposable chambers Standard Count (Leja, Amsterdam, Holland), Cellvision (Anthos-ec, Heerhugowaard, The Netherlands), and Cell Vu (Fertility Technologies, Inc. Natick, MA) showed small variation and correct estimation of bead concentration. All the reusable chambers gave relatively large variability, with tendency to underestimate (improved Neubauer; Hawksley, Lancing, United Kingdom) or to overestimate the bead concentration. The use of plunger-displacement pipettes resulted in an overestimation of bead concentration in medium but not in seminal plasma. Counting the sperm suspension using plunger-displacement pipettes indicated that the Bürker hemocytometer overestimated concentration relative to that obtained by Cellvision and Makler Counting Chambers (Sefi Medical Instruments, Haifa, Israel) and that the improved Neubauer presented the lowest variability (7.1%). Conclusion(s): The improved Neubauer hemocytometer is the standard for sperm counting, though disposable chambers give reliable results as well. If beads are used to evaluate the accuracy of counting chambers, it is recommended to dilute them with seminal plasma.

Quantitative studies of coccidial development in miniature cultures of animal cells

The use of miniature culture chambers reduces the time required to count intracellular coccidia in standard areas of infected monolayer cell cultures. The saving in time allows an increased number of replicate cultures, thereby producing a 50% reduction in standard error. We use a commercially available non-plastic device which (a) provides easily cleaned, reusable chambers, (b) allows easy fixing and staining of culture slides which (c) can be examined at any magnification under a light microscope and (d) can be retained as a permanent record.

A chamber for organotypic culture; adapted for growing large volumes of tissue.

Plastic vessels, one glued within the other, form a reusable chamber for the culture of large volumes (3-5 mm3) of tissue minced into fine particles (0.25 mm or less). The vessels are three in number: one for water to prevent dehydration, the second for the medium and the third for tissue. The medium is passed from its vessel to the tissue by means of an absorbent cotton wick. Plastic tubing, ventilating the chamber, opens to the outside and may be utilized for the exchange of gasses or for replenishing the culture with a desired medium. Any oven used for tissue culture may be fitted with jets for the supply of gasses to the chamber, thus allowing a number of chambers to function concomitantly. Hypophyseal intermediate lobe tissue as well as mature and embryonic tissue from many species have been successfully grown in the chamber, and it can be utilized also for the cultivation of viruses, for the the study of antibody formation in various organs and for biochemical investigations.