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Methods in Molecular Biology (Clifton,... 2018Here we describe a culture technique of cells dissociated from the external muscularis of the guinea pig small intestine, which allows us to maintain all the elements...
Here we describe a culture technique of cells dissociated from the external muscularis of the guinea pig small intestine, which allows us to maintain all the elements involved in the intestinal peristaltic reflex. After a few days in culture, these cells reorganize to form a small group of cells that permit the generation of pacemaker activity, spontaneous contractions, and the development of inhibitory and excitatory junction potentials in the petri dish, all elements involved in the peristaltic reflex. Therefore, these co-cultures are suitable to study the cellular and molecular aspects related to the development, maintenance, and modulation of motor intestinal functions.
Topics: Action Potentials; Animals; Coculture Techniques; Female; Guinea Pigs; Intestine, Small; Male; Mice, Inbred C57BL; Motor Neurons; Muscle Contraction; Myocytes, Smooth Muscle; Peristalsis; Rats
PubMed: 29222802
DOI: 10.1007/978-1-4939-7571-6_34 -
Heliyon Jan 2019The overall structure of dihydrate cupric chloride (CuCl * 2HO) crystallization patterns in the presence of bovine serum albumin (BSA) in a Petri dish is influenced by...
The overall structure of dihydrate cupric chloride (CuCl * 2HO) crystallization patterns in the presence of bovine serum albumin (BSA) in a Petri dish is influenced by dewetting. The dewetting behavior, which can be either before or after initial CuCl nucleation, depends on the amount of CuCl₂ and BSA in the Petri dish. We postulate that the concentration and/or temperature gradient area in the dish, which is built up during the evaporation process, coincides with the location where dewetting predominantly starts. This hypothesis could be supported by measurements of the CuCl coverage of the Petri dish. During the evaporation the height of the meniscus at the rim of the Petri dish recedes in favor of the central Petri dish area. This could not be explained by the above mentioned hypothesis.
PubMed: 30627687
DOI: 10.1016/j.heliyon.2018.e01102 -
British Journal of Nursing (Mark Allen...
Topics: Academies and Institutes; Animals; Attitude to Health; Humans; Licensure; Public Opinion; Research Embryo Creation; Stem Cell Transplantation; Transplantation Chimera; United Kingdom
PubMed: 18414276
DOI: 10.12968/bjon.2008.17.2.28131 -
Andrologia Dec 2022Herein, we introduced a novel individual sperm freezing device named SpermCD, which consists of a right angular cryopiece (RA-Cryopiece, or "C") and a grooved petri dish...
Herein, we introduced a novel individual sperm freezing device named SpermCD, which consists of a right angular cryopiece (RA-Cryopiece, or "C") and a grooved petri dish ("D"). SpermCD allows embryologists to transfer sperm and perform ICSI on the same focal plane. Thirty-five patients underwent single sperm cryopreservation using SpermCD, including four patients with non-obstructive azoospermia (NOA), 14 patients with virtual azoospermia and 17 patients with cryptozoospermia. One hundred and twenty-five cryopreserved spermatozoa from nine patients were thawed on the day of the oocyte retrieval and 121 spermatozoa were found, with a sperm recovery rate of 97.1 ± 4.6%. Sixty-five MII oocytes from their spouse were injected with thawed sperm. Normal fertilization and high-quality embryo rates were 68.0% ± 33.2% and 24.4% ± 22.2%. Nineteen transplantable embryos were formed after fertilization with frozen sperm, eight of which were transplanted in five couples, resulting in four successful deliveries. SpermCD is a simple and practical individual sperm freezing device.
Topics: Humans; Male; Azoospermia; Sperm Injections, Intracytoplasmic; Freezing; Embryo Transfer; Spermatozoa; Cryopreservation; Testis
PubMed: 36285609
DOI: 10.1111/and.14619 -
Journal of Neuroscience Methods Jan 2018Microscope chambers that accept glass coverslips with cultured cells are often used to monitor intracellular Ca concentration ([Ca]) during cell superfusion....
BACKGROUND
Microscope chambers that accept glass coverslips with cultured cells are often used to monitor intracellular Ca concentration ([Ca]) during cell superfusion. Unfortunately, the experimental maneuvers associated with the coverslip installation in these chambers (medium removal and re-application) trigger unintended [Ca] elevations.
NEW METHOD
To prevent these [Ca] elevations, a Petri dish insert has been constructed. The insert features a superfusion-optimized well to grow cell cultures. After this insert is removed from the Petri dish, the well retains the medium. This feature allows the inserts to be installed in microscope chambers while keeping the cells submerged at all times.
RESULTS
These inserts were used to test the impact of a transient medium removal from the well (an equivalent of a coverslip removal from the medium) on [Ca] in primary murine cortical neurons and astrocytes, and in HEK-293 cells. In all of these models, the medium removal/re-application caused a micromolar [Ca] spike. While in neurons this spike was caused by a Ca influx, in astrocytes and HEK-293 cells, it was caused by a Ca release from intracellular stores. After the spike, a subpopulation of neurons failed to restore low [Ca]; in 24% of the astrocytes, the spike triggered [Ca] oscillations. However, prior to the spike, [Ca] was low and uniform in all these cells.
COMPARISON WITH EXISTING METHOD(S)
The new method avoids the artificially-induced [Ca] elevations that take place during the handling of glass coverslips with cultured cells.
CONCLUSIONS
The new method allows monitoring [Ca] without disturbing the basal [Ca] levels.
Topics: Animals; Astrocytes; Calcium Signaling; Cell Culture Techniques; Coculture Techniques; Culture Media; Female; HEK293 Cells; Humans; Male; Mice, Inbred C57BL; Neurons; Optical Imaging; Primary Cell Culture
PubMed: 29132810
DOI: 10.1016/j.jneumeth.2017.11.007 -
Lab on a Chip May 2009We report high-speed real-time PCR performed on an unmodified disposable polystyrene Petri dish. The reaction cycle relies solely on an infrared laser for heating; no...
We report high-speed real-time PCR performed on an unmodified disposable polystyrene Petri dish. The reaction cycle relies solely on an infrared laser for heating; no conventional heater is required. Nanoliter droplets of PCR mixture as water-in-oil emulsions printed in an array format served as individual PCR microreactors. A simple contact printing technique was developed to generate a large array of uniform sized nanoliter droplets using disposable pipette tips. Printed droplets showed variation of less than 10% in volume and the oil/water/polystyrene interface formed a compact droplet microreactor approximately spherical in shape. The uniform droplet array was used to optimize the laser power required for the two heating steps of PCR, annealing/extension and melting, while the ambient conditions were at room temperature. The optical heating allows for an extremely fast heating rate due to the selective absorption of the infrared laser by PCR buffer only and not the oil or polystyrene Petri dish, allowing completion of 40 amplification cycles in approximately 6 minutes. The quantitative assay capability of the system is also presented and discussed.
Topics: Computer-Aided Design; Equipment Design; Equipment Failure Analysis; Heating; Lasers; Microchemistry; Microfluidic Analytical Techniques; Nanotechnology; Oligonucleotide Array Sequence Analysis; Polymerase Chain Reaction; Reproducibility of Results; Sensitivity and Specificity
PubMed: 19370241
DOI: 10.1039/b817288a -
Postdoc Journal : a Journal of... Nov 2015Ohio State University researchers have made a leap forward in disease research by creating an eraser sized human "brain" in a petri dish. Although lacking a circulatory...
Ohio State University researchers have made a leap forward in disease research by creating an eraser sized human "brain" in a petri dish. Although lacking a circulatory system their brain model includes spinal cord, cortex, midbrain, brain stem, and even the beginnings of an eye- aiding in the effectiveness of research on complex neurological disease. To create their new brain model, the researchers converted adult skin cells into pluripotent stem cells, which afforded the opportunity to build the multiple nervous cell types required for such a complex system. Having this tissue model will assist researchers in developing new disease models, and thus, facilitate the development of novel clinical interventions.
PubMed: 27429994
DOI: No ID Found -
Journal of Visualized Experiments : JoVE Feb 2021To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate their food...
To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate their food environment. The fruit fly, Drosophila melanogaster, is a genetically tractable model organism that is widely used to decipher the molecular, cellular, and neural underpinnings of food preference. To analyze fly food preference, a robust feeding method is needed. Described here is a two-choice feeding assay, which is rigorous, cost-saving, and fast. The assay is Petri-dish-based and involves the addition of two different foods supplemented with blue or red dye to the two halves of the dish. Then, ~70 prestarved, 2-4-day-old flies are placed in the dish and allowed to choose between blue and red foods in the dark for about 90 min. Examination of the abdomen of each fly is followed by the calculation of the preference index. In contrast to multiwell plates, each Petri dish takes only ~20 s to fill and saves time and effort. This feeding assay can be employed to quickly determine whether flies like or dislike a particular food.
Topics: Animals; Biological Assay; Coloring Agents; Drosophila melanogaster; Feeding Behavior; Food Preferences; Indicators and Reagents; Starvation
PubMed: 33645577
DOI: 10.3791/62051 -
Frontiers in Microbiology 2021Cocultivation is an emerging and potential way to investigate microbial interaction in the laboratory. Extensive researches has been carried out over the years, but some...
Cocultivation is an emerging and potential way to investigate microbial interaction in the laboratory. Extensive researches has been carried out over the years, but some microorganism cocultivation are not easy to implement in the laboratory, especially the fungus-fungus (FF) cocultivation, owing to the obstacles such as fungal different growth rate, limited growing space, hyphae intertwining, and difficulty of sample separation, etc. In this research, a double-sided petri dish (DSPD) was designed and carried out as a tool to study FF cocultivation in the laboratory. A natural FF cocultivation of spp. and inspired from black-skin-red-koji (BSRK), were studied. By using DSPD, the aforementioned obstacles in the FF cocultivation study were overcome through co-culturing spp. and on each side of DSPD. The characteristics of monocultured and co-cultured spp. and were compared and analyzed, including colonial and microscopic morphologies, and main secondary metabolites (SMs) of spp. analyzed by high performance liquid chromatography. And a novel SM was found to be produced by M7 when co-cultured with CBS 513.88. Since the above mentioned obstacles, were overcome, we obtained good quality of transcriptome data for further analysis. These results indicate that DSPD might be an efficient tool for investigation of microbial interaction, in particular, for FF interaction.
PubMed: 34177849
DOI: 10.3389/fmicb.2021.670684 -
Lab on a Chip Feb 2021The selection of high quality sperm is critical for intracytoplasmic sperm injection (ICSI), a prevalent assisted reproduction technology. However, standard selection...
The selection of high quality sperm is critical for intracytoplasmic sperm injection (ICSI), a prevalent assisted reproduction technology. However, standard selection methods are time-consuming and fail to recover the most viable sperm, thereby limiting the ICSI success rate. Microfluidics enables rapid selection of viable sperm in a manner representing in vivo processes, however, existing platforms lack clinical applicability. Here, we present FertDish, which integrates the clinically established ICSI Petri dish with a film featuring an array of sperm-selecting microchannels for selection of sperm directly from semen. The FertDish format mimics the clinician-familiar ICSI dish setup, and provides rapid (<10 min) single stage sperm preparation that circumvents standard labour-intensive multi-stage sperm processing steps. Tests with human donor and patient semen samples show that FertDish enables the selection of a high quality sperm sub-population, featuring improvements in DNA fragmentation index of more than 91% (donor) and 74% (patient) versus raw semen and 50% (donor) and 63% (patient) versus standard methods, and a distribution of more than 97% sperm with viable and high level DNA. The FertDish enables a high sperm recovery rate (>3.3 × 105 sperm per mL), and is readily adaptable to the clinical workflow with potential to improve ICSI outcomes.
Topics: Humans; Male; Microfluidics; Sperm Injections, Intracytoplasmic; Spermatozoa
PubMed: 33507191
DOI: 10.1039/d0lc00874e