Facilitated crystal handling using a simple device for evaporation reduction in microtiter plates.
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ABSTRACT:
SUBMITTER: Barthel T
PROVIDER: S-EPMC7941301 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Formulation screening for biotherapeutics can cover a vast array of excipients and stress conditions. These studies consume quantities of limited material and, with higher concentrated therapeutics, more material is needed. Here, we evaluate the use of crystal zenith (CZ) microtiter plates in conjunction with FluoroTec-coated butyl rubber mats as a small-volume, high-throughput system for formulation stability studies. The system was studied for evaporation, edge effects, and stability with comparisons to type 1 glass and CZ vials for multiple antibodies and formulations. Evaporation was minimal at 4°C and could be reduced at elevated temperatures using sealed, mylar bags. Edge effects were not observed until 12 weeks at 40°C. The overall stability ranking as measured by the rate of change in high molecular weight and percent main peak species was comparable across both vials and plates at 4°C and 40°C out to 12 weeks. Product quality attributes as measured by the multi-attribute method were also comparable across all containers for each molecule formulation. A potential difference was measured for subvisible particle analysis, with the plates measuring lower particle counts than the vials. Overall, CZ plates are a viable alternative to traditional vials for small-volume, high-throughput formulation stability screening studies.
| S-EPMC6941220 | biostudies-literature
Project description:BackgroundConventional crop protection has major drawbacks, such as developing pest and pathogen insensitivity to pesticides and low environmental compatibility. Therefore, alternative crop protection strategies are needed. One promising approach treats crops with chemical compounds that induce the primed state of enhanced defense. However, identifying priming compounds is often tedious as it requires offline sampling and analysis. High throughput screening methods for the analysis of priming-active compounds have great potential to simplify the search for such compounds. One established method to identify priming makes use of parsley cell cultures. This method relies on measurement of fluorescence of furanocoumarins in the final sample. This study demonstrates for the first time the online measurement of furanocoumarins in microtiter plates. As not all plants produce fluorescence molecules as immune response, a signal, which is not restricted to a specific plant is required, to extend online screening methods to other plant cell cultures. It was shown that the breathing activity of primed parsley cell cultures increases, compared to unprimed parsley cell cultures. The breathing activity can by monitored online. Therefore, online identification of priming-inducing compounds by recording breathing activity represents a promising, straight-forward and highly informative approach. However, so far breathing has been recorded in shake flasks which suffer from low throughput. For industrial application we here report a high-throughput, online identification method for identifying priming-inducing chemistry.ResultsThis study describes the development of a high-throughput screening system that enables identifying and analyzing the impact of defense priming-inducing compounds in microtiter plates. This screening system relies on the breathing activity of parsley cell cultures. The validity of measuring the breathing activity in microtiter plates to drawing conclusions regarding priming-inducing activity was demonstrated. Furthermore, for the first time, the fluorescence of the priming-active reference compound salicylic acid and of furanocoumarins were simultaneously monitored online. Dose and time studies with salicylic acid-treated parsley cell suspensions revealed a wide range of possible addition times and concentrations that cause priming. The online fluorescence measuring method was further confirmed with three additional compounds with known priming-causing activity.ConclusionsDetermining the OTR, fluorescence of the priming-active chemical compound SA and of furanocoumarins in parsley suspension cultures in MTPs by online measurement is a powerful and high-throughput tool to study possible priming compounds. It allows an in-depth screening for priming compounds and a better understanding of the priming process induced by a given substance. Evaluation of priming phenomena via OTR should also be applicable to cell suspensions of other plant species and varieties and allow screening for priming-inducing chemical compounds in intact plants. These online fluorescence methods to measure the breathing activity, furanocoumarin and SA have the potential to accelerate the search for new priming compounds and promote priming as a promising, eco-friendly crop protection strategy.
| S-EPMC9893529 | biostudies-literature
Project description:Paper-based analytical devices have prominently emerged as a group of diagnostic tools with prospective to eliminate the expensive, time-consuming, and intricate analytical methodologies. Wax printing has been a dominant technique to fabricate hydrophobic patterns on paper for fluid control, but the discontinuation of commercial solid ink printers has begun a genesis of alternate wax patterning strategies. In this study, a simple and rapid fabrication methodology for realizing a 96-well microtiter plate on paper has been developed. The method involves the use of commercially available polystyrene microplates as a stamp for wax patterning. The technique further eradicates the requirement of customized stamps and the step of heating paper substrates for creating wax barriers. Thus, wax stamped paper microplates can be used for a wide range of bioanalytical tests maneuvering reduced generation of non-biodegradable waste, minimal reagent usage, and inexpensive readout strategies. The viability of the fabricated platform has been assessed by colorimetric detection of glutathione using 3,3′,5,5′-tetramethylbenzidine–H2O2 redox system. RGB analysis of the colorimetric response showed a linear concentration range from 0 to 90 µM (R2 = 0.989) along with a detection limit of 28.375 µM. Supplementary Information The online version contains supplementary material available at 10.1007/s10404-022-02606-3.
| S-EPMC9632569 | biostudies-literature
Project description:Interfacial water evaporation technology by using solar energy provides one of the promising pathways for freshwater shortage management. However, current research mainly focuses on the improvement of evaporation efficiency by macro or microregulations, ignoring the steam temperature, which is a manifestation of the quality of water. Herein not only is a high-rate solar evaporation achieved but also steam temperature is enhanced by a simple three-tier (wet absorber-air gap-dry absorber) device. In a routine interfacial evaporation test, the evaporator achieves a stable evaporation rate up to 2.15 kg m-2 h-1 under one sun, demonstrating a competitive evaporation rate compared with other reports. With the three-tier device, the steam temperature can increase 33.7%, 41.13%, and 47% without dry absorber under one sun, two sun, and three sun illumination, respectively. At the same time, the steam temperature can be as high as 95.5 °C under three sun intensities. This work provides the possibility of using a simple three-tier device for high-temperature steam generation without extra energy input, which contributes to an idea for future research on the production high-quality water.
| S-EPMC8025398 | biostudies-literature
Project description:BackgroundLight transmission aggregometry (LTA) can be performed with microtiter plates (96-well LTA). When conducting LTA, an agonist is added to platelet-rich plasma and the sample is shaken for minutes after which absorbance readings are done. Platelet aggregation is detected as decrease in absorbance. However, the classical method is cumbersome and therefore microtiter plates can be used for concomitant testing of multiple samples. Furthermore, it would be convenient to prepare the plate in advance of platelet aggregation testing. Aim: The aim of the present study was to establish a simplified 96-well LTA protocol, where plates were pre-coated with agonists and stored at -80 C until use.ResultsWe developed and validated a protocol for 96-well LTA using a Victor X5 plate reader and pre-coated microtiter plates. The minimum requirement of platelet-rich plasma was 45 ?L per sample and the sample platelet count should not be below 100 x109/L. Optimal absorbance reading was 595 nm wavelengths. Platelet aggregation results were higher at 37°C than at room temperature. Platelet adherence to wells after stimulation was observed; it was not avoided by pre-coating of the wells with gelatin. A range of up to 7 concentrations for each agonist (collagen, arachidonic acid, adenosine diphosphate, thrombin receptor-activating peptide and protease-activated receptor-4) was tested concomitantly. A transient rise in platelet aggregation was observed after 2 minutes of shaking in some samples with low agonist concentration, and platelet aggregation was optimal after 10 minutes of shaking for samples with high agonist concentration. Plates could be stored at -80°C for 15 days without significant change in the platelet aggregation results.ConclusionThe 96-well LTA is suitable for platelet aggregation testing and a range of agonist concentrations can be concomitantly tested.
| S-EPMC5638243 | biostudies-literature
Project description:Droplet-based microfluidic screening techniques can benefit from interfacing established microtiter plate-based screening and sample management workflows. Interfacing tools are required both for loading preconfigured microtiter-plate (MTP)-based sample collections into droplets and for dispensing the used droplets samples back into MTPs for subsequent storage or further processing. Here, we present a collection of Digital Microfluidic Pipetting Tips (DMPTs) with integrated facilities for droplet generation and manipulation together with a robotic system for its operation. This combination serves as a bidirectional sampling interface for sample transfer from wells into droplets (w2d) and vice versa droplets into wells (d2w). The DMPT were designed to fit into 96-deep-well MTPs and prepared from glass by means of microsystems technology. The aspirated samples are converted into the channel-confined droplets' sequences separated by an immiscible carrier medium. To comply with the demands of dose-response assays, up to three additional assay compound solutions can be added to the sample droplets. To enable different procedural assay protocols, four different DMPT variants were made. In this way, droplet series with gradually changing composition can be generated for, e.g., 2D screening purposes. The developed DMPT and their common fluidic connector are described here. To handle the opposite transfer d2w, a robotic transfer system was set up and is described briefly.
| S-EPMC9316479 | biostudies-literature
Project description:We have developed a methodology that enables for the rapid measurement of ionization constants (pK a) of series of compounds by UV spectrophotometry. This protocol, which is straightforward to set up, takes advantage of the sensitivity of UV spectroscopy and the throughput enabled by the 96-well microplate (as opposed to the use of 1 cm quartz cuvette). The compounds, in stock solutions in DMSO, are dissolved in several aqueous buffer solutions directly in the microtiter plate, allowing the simultaneous determination of the UV spectra as a function of pH. Further treatment of the data provides the pK a values in a medium-throughput manner. The pK a values of 11 new antitrypanosomal dibasic compounds were determined using this methodology.
| S-EPMC4027549 | biostudies-literature
Project description:Affinity ligands such as antibodies are widely used in (bio)medical research for purifying proteins from complex biological samples. These ligands are generally immobilized onto solid supports which facilitate the separation of a captured protein from the sample matrix. Adsorptive microtiter plates are commonly used as solid supports prior to immunochemical detection (e.g., immunoassays) but hardly ever prior to liquid chromatography-mass spectrometry (LC-MS-)-based detection. Here, we describe the use of adsorptive microtiter plates for protein enrichment prior to LC-MS detection, and we discuss opportunities and challenges of corresponding workflows, based on examples of targeted (i.e., soluble receptor for advanced glycation end-products (sRAGE) in human serum) and discovery-based workflows (i.e., transcription factor p65 (NF-κB) in lysed murine RAW 264.7 macrophages and peptidyl-prolyl cis-trans isomerase FKBP5 (FKBP5) in lysed human A549 alveolar basal epithelial cells). Thereby, we aim to highlight the potential usefulness of adsorptive microtiter plates in affinity purification workflows prior to LC-MS detection, which could increase their usage in mass spectrometry-based protein research.
| S-EPMC8576839 | biostudies-literature
Project description:One of the main trends in the development of solar interface evaporation technology is the simple, efficient, and environmentally friendly bio-based evaporation device. However, the development of bio-based evaporators with high water evaporation rates and good pollution removal capability is a significant challenge. Here, we present a carbonized cattail-agarose hydrogel (CCAH) membrane with numerous microchannels resembling bamboo knots, exceptional hydrophilicity, outstanding light absorption capability, and potent adsorption. Under one solar irradiation, its evaporation rate and efficiency reached 1.93 kg m-2 h-1 and 95.8%, respectively. More importantly, the CCAH membrane produces steam water that is almost totally free of salts (Na+, K+, Mg2+, and Ca2+), heavy metal ions (Pb2+, Cd2+, and Cr2+), and organic dyes (Rhodamine B, methylene blue, and methyl orange). The CCAH membrane is highly promising for the use of saltwater desalination and wastewater recovery to help people in impoverished areas with water scarcity problems.
| S-EPMC9692801 | biostudies-literature
Project description:Microtiter plate-based bacterial biofilm assay is frequently used to study bacterial biofilm development and growth. While this assay is simple and relatively high-throughput, it frequently shows difficulty in establishing robust biofilm attachment in the wells. We report that the consistency of bacterial biofilm assays carried out in microtiter plates subjected to abrasive treatment, by sandblasting or drill press grinding, is significantly improved in a Pseudomonas fluorescens Pf0-1 model. Scanning electron microscopy imaging suggests that the treated surfaces could provide points of attachment to facilitate the recruitment of bacteria in the initial phase of biofilm colony establishment. The sandblast treated polypropylene, but not polystyrene, plates were found suitable in studying the impact of flavonoid quercetin on the biofilm formation in Bacillus subtilis FB17. Further investigation revealed that due to the hydrophobicity of the polystyrene surfaces, a greater amount of quercetin was adsorbed on the plate surface, effectively lowering the concentration of the flavonoid in solution.
| S-EPMC9086168 | biostudies-literature