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Sensors (Basel, Switzerland) Jan 2024In recent years, the demand for effective intracytoplasmic sperm injection (ICSI) for the treatment of male infertility has increased. The ICSI operation is complicated...
In recent years, the demand for effective intracytoplasmic sperm injection (ICSI) for the treatment of male infertility has increased. The ICSI operation is complicated as it involves delicate organs and requires a high level of skill. Several cell manipulation systems that do not require such skills have been proposed; notably, several automated methods are available for cell rotation. However, these methods are unfeasible for the delicate ICSI medical procedure because of safety issues. Thus, this study proposes a microscopic system that enables intuitive micropipette manipulation using a haptic device that safely and efficiently performs the entire ICSI procedure. The proposed system switches between field-of-view expansion and three-dimensional image presentation to present images according to the operational stage. In addition, the system enables intuitive pipette manipulation using a haptic device. Experiments were conducted on microbeads instead of oocytes. The results confirmed that the time required for the experimental task was improved by 52.6%, and the injection error was improved by 75.3% compared to those observed in the conventional system.
Topics: Humans; Male; Sperm Injections, Intracytoplasmic; Haptic Interfaces; Semen; Infertility, Male; Oocytes; Spermatozoa
PubMed: 38276402
DOI: 10.3390/s24020711 -
PloS One 2024In remote communities, diagnosis of G6PD deficiency is challenging. We assessed the impact of modified test procedures and delayed testing for the point-of-care...
In remote communities, diagnosis of G6PD deficiency is challenging. We assessed the impact of modified test procedures and delayed testing for the point-of-care diagnostic STANDARD G6PD (SDBiosensor, RoK), and evaluated recommended cut-offs. We tested capillary blood from fingerpricks (Standard Method) and a microtainer (BD, USA; Method 1), venous blood from a vacutainer (BD, USA; Method 2), varied sample application methods (Methods 3), and used micropipettes rather than the test's single-use pipette (Method 4). Repeatability was assessed by comparing median differences between paired measurements. All methods were tested 20 times under laboratory conditions on three volunteers. The Standard Method and the method with best repeatability were tested in Indonesia and Nepal. In Indonesia 60 participants were tested in duplicate by both methods, in Nepal 120 participants were tested in duplicate by either method. The adjusted male median (AMM) of the Biosensor Standard Method readings was defined as 100% activity. In Indonesia, the difference between paired readings of the Standard and modified methods was compared to assess the impact of delayed testing. In the pilot study repeatability didn't differ significantly (p = 0.381); Method 3 showed lowest variability. One Nepalese participant had <30% activity, one Indonesian and 10 Nepalese participants had intermediate activity (≥30% to <70% activity). Repeatability didn't differ significantly in Indonesia (Standard: 0.2U/gHb [IQR: 0.1-0.4]; Method 3: 0.3U/gHb [IQR: 0.1-0.5]; p = 0.425) or Nepal (Standard: 0.4U/gHb [IQR: 0.2-0.6]; Method 3: 0.3U/gHb [IQR: 0.1-0.6]; p = 0.330). Median G6PD measurements by Method 3 were 0.4U/gHb (IQR: -0.2 to 0.7, p = 0.005) higher after a 5-hour delay compared to the Standard Method. The definition of 100% activity by the Standard Method matched the manufacturer-recommended cut-off for 70% activity. We couldn't improve repeatability. Delays of up to 5 hours didn't result in a clinically relevant difference in measured G6PD activity. The manufacturer's recommended cut-off for intermediate deficiency is conservative.
Topics: Humans; Male; Glucosephosphate Dehydrogenase; Pilot Projects; Sodium Oxybate; Glucosephosphate Dehydrogenase Deficiency; Biosensing Techniques
PubMed: 38241389
DOI: 10.1371/journal.pone.0296708 -
Scientific Reports Jan 2024The patch-clamp technique has revolutionized neurophysiology by allowing to study single neuronal excitability, synaptic connectivity, morphology, and the transcriptomic...
The patch-clamp technique has revolutionized neurophysiology by allowing to study single neuronal excitability, synaptic connectivity, morphology, and the transcriptomic profile. However, the throughput in recordings is limited because of the manual replacement of patch-pipettes after each attempt which are often also unsuccessful. This has been overcome by automated cleaning the tips in detergent solutions, allowing to reuse the pipette for further recordings. Here, we developed a novel method of automated cleaning by sonicating the tips within the bath solution wherein the cells are placed, reducing the risk of contaminating the bath solution or internal solution of the recording pipette by any detergent and avoiding the necessity of a separate chamber for cleaning. We showed that the patch-pipettes can be used consecutively at least ten times and that the cleaning process does not negatively impact neither the brain slices nor other patched neurons. This method, combined with automated patch-clamp, highly improves the throughput for single and especially multiple recordings.
Topics: Ultrasonics; Detergents; Neurons; Neurophysiology; Patch-Clamp Techniques
PubMed: 38238544
DOI: 10.1038/s41598-024-51837-7 -
Analytica Chimica Acta Feb 2024In "shotgun" approaches involving high-performance liquid chromatography or capillary zone electrophoresis (CZE), matrix removal prior to sample analysis is considered...
BACKGROUND
In "shotgun" approaches involving high-performance liquid chromatography or capillary zone electrophoresis (CZE), matrix removal prior to sample analysis is considered as an indispensable tool. Despite the fact that CZE offers a high tolerance towards salts, most publications reported on the use of desalting. There seems to be no clear consensus on the utilization of desalting in the CZE-MS community, most probably due to the absence of works addressing the comparison of desalted and non-desalted digests. Our aim was to fill this research gap using protein samples of varying complexity in different sample matrices.
RESULTS
First, standard protein digests were analyzed to build the knowledge on the effect of sample clean-up by solid-phase extraction (SPE) pipette tips and the possible stacking phenomena induced by different sample matrices. Desalting led to a somewhat altered peptide profile, the procedure affected mostly the hydrophilic peptides (although not to a devastating extent). Nevertheless, desalting samples allowed remarkable stacking efficiency owing to their low-conductivity sample background, enabling a so-called field-amplified sample stacking phenomenon. Non-desalted samples also produced a stacking event, the mechanism of which is based on transient-isotachophoresis due to the presence of high-mobility ions in the digestion buffer itself. Adding either extra ammonium ions or acetonitrile into the non-desalted digests enhanced the stacking efficiency. A complex sample (yeast cell lysate) was also analyzed with the optimal conditions, which yielded similar tendencies.
SIGNIFICANCE
Based on these results, we propose that sample clean-up in the bottom-up sample preparation process prior to CZE-MS analysis can be omitted. The preclusion of desalting can even enhance detection sensitivity, separation efficiency or sequence coverage.
Topics: Peptide Mapping; Tandem Mass Spectrometry; Proteomics; Electrophoresis, Capillary; Peptides; Ions
PubMed: 38220294
DOI: 10.1016/j.aca.2023.342162 -
Botanical Studies Jan 2024The bimolecular fluorescence complementation (BiFC) assay is commonly used for investigating protein-protein interactions. While several BiFC detection systems have been...
BACKGROUND
The bimolecular fluorescence complementation (BiFC) assay is commonly used for investigating protein-protein interactions. While several BiFC detection systems have been developed, there is a limited amount of research focused on using laser scanning confocal microscope (LSCM) techniques to observe protoplasts. Protoplasts are more susceptible to damage and instability compared to their original cell state due to the preparation treatments they undergo, which makes it challenging for researchers to manipulate them during observation under LSCMs. Therefore, it is crucial to utilize microscope techniques properly and efficiently in BiFC assays.
RESULTS
When the target fluorescence is weak, the autofluorescence of chloroplast particles in protoplasts can interfere with the detection of BiFC signals localized in the nuclear region. Spectrum analysis revealed that chloroplast autofluorescence can be excited by lasers of various types, with the highest fluorescence signal observed at around 660 nm. Furthermore, our investigation into the impact of different pipette tips on the integrity of protoplast samples indicated that the utilization of cut tips with larger openings can mitigate cell breakage. We presented a workflow of LSCM techniques for investigating protoplast BiFC and discussed the microscopic manipulation involved in sample preparation and image capturing.
CONCLUSION
When the BiFC signals are weak, they may be affected by chloroplast autofluorescence. However, when used properly, the autofluorescence of chloroplasts can serve as an excellent internal marker for effectively distinguishing other signals. In combination with other findings, this study can provide valuable reference for researchers conducting BiFC assays and related studies.
PubMed: 38194078
DOI: 10.1186/s40529-024-00409-z -
BioTechniques Feb 2024Additive manufacturing (3D printing) has been deployed across multiple platforms to fabricate bioengineered tissues. We demonstrate the use of a Thermal Inkjet Pipette...
Additive manufacturing (3D printing) has been deployed across multiple platforms to fabricate bioengineered tissues. We demonstrate the use of a Thermal Inkjet Pipette System (TIPS) for targeted delivery of cells onto manufactured substrates to design bio-bandages. Two cell lines - HEK 293 (kidney) and K7M2 wt (bone) - were applied using TIPS. We demonstrate a novel means for targeted cell delivery to a hydrogel support structure. These cell/support constructs (bio-bandages) had a high viability for survival and growth over extended periods. Combining a flexible biosupport with application of cells via TIPS printing now for the first time allows for custom cell substrate constructs with various densities to be deployed for regenerative medicine applications.
Topics: Humans; Hydrogels; Tissue Engineering; HEK293 Cells; Printing, Three-Dimensional; Tissue Scaffolds; Bioprinting
PubMed: 38084384
DOI: 10.2144/btn-2023-0052 -
European Journal of Human Genetics :... Apr 2024News stories and patient-facing material about genetic tests are often illustrated by images, but the content of such images and the messages they propagate are rarely...
News stories and patient-facing material about genetic tests are often illustrated by images, but the content of such images and the messages they propagate are rarely scrutinised. Stock image banks were searched to identify a hundred images relating to genetic tests and analysed using a multimodal critical discourse approach, aiming to identify what the images featured, how they were composed, and what they communicated about genetic testing. We found that images tended to focus on technical aspects of sample processing (for example, pipetting) and drew on older technologies (for example slab gel electrophoresis) when representing data arising from genetic tests. Composition choices like focussing images around pipette tips, or emphasising colour or brightness of electrophoretic bands, represented genetic testing as precise, unambiguous and illuminating. Only 7% of images featured a person having a genetic test, and only one image alluded to communication of genetic results. Current popular visual representations of genetic testing rarely highlight the possibility of uncertain or non-diagnostic outcomes, and may contribute to high public expectations of informativeness and certainty from such tests.
Topics: Humans; Genetic Testing; Gels
PubMed: 38066171
DOI: 10.1038/s41431-023-01508-4 -
Environmental Health Perspectives Dec 2023The rapid evolution of electronic cigarette (e-cigarette) products warrants surveillance of the differences in exposure across device types-modifiable devices (MODs),...
BACKGROUND
The rapid evolution of electronic cigarette (e-cigarette) products warrants surveillance of the differences in exposure across device types-modifiable devices (MODs), cartridge ("pod")-containing devices (PODs), disposable PODs (d-PODs)-and flavors of the products available on the market.
OBJECTIVE
This study aimed to measure and compare metal aerosol concentrations by device type and common flavors.
METHODS
We collected aerosol from 104 MODs, 67 PODs (four brands: JUUL, Bo, Suorin, PHIX), and 23 d-PODs (three brands: ZPOD, Bidi, Stig) via droplet deposition in a series of conical pipette tips. Metals and metalloids [aluminum (Al), arsenic (As), cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), lead (Pb), antimony (Sb), tin (Sn), and zinc (Zn)] were measured using inductively coupled plasma mass spectrometry (ICP-MS), results were log-transformed for statistical analysis, and concentrations are reported in aerosol units ().
RESULTS
Of the 12 elements analyzed, concentrations were statistically significantly higher in MOD devices, except for Co and Ni, which were higher in PODs and d-PODs. Of the POD brands analyzed, PHIX had the highest median concentrations among four metals (Al, Ni, Pb, and Sn) compared to the rest of the POD brands. According to POD flavor, seven metals were three to seven orders of magnitude higher in tobacco-flavored aerosol compared to those in mint and mango flavors. Among the d-POD brands, concentrations of four metals (Al, Cu, Ni, and Pb) were higher in the ZPOD brand than in Bidi Stick and Stig devices. According to d-POD flavor, only Cr concentrations were found to be statistically significantly higher in mint than tobacco-flavored d-PODs.
DISCUSSION
We observed wide variability in aerosol metal concentrations within and between the different e-cigarette device types, brands, and flavors. Overall, MOD devices generated aerosols with higher metal concentrations than PODs and d-PODs, and tobacco-flavored aerosols contained the highest metal concentrations. Continued research is needed to evaluate additional factors (i.e., nicotine type) that contribute to metal exposure from new and emerging e-cigarette devices in order to inform policy. https://doi.org/10.1289/EHP11921.
Topics: Electronic Nicotine Delivery Systems; Lead; Aluminum; Aerosols; Copper; Chromium
PubMed: 38048100
DOI: 10.1289/EHP11921 -
Digital Discovery Oct 2023Closed-loop experiments can accelerate material discovery by automating both experimental manipulations and decisions that have traditionally been made by researchers....
Closed-loop experiments can accelerate material discovery by automating both experimental manipulations and decisions that have traditionally been made by researchers. Fast and non-invasive measurements are particularly attractive for closed-loop strategies. Viscosity is a physical property for fluids that is important in many applications. It is fundamental in application areas such as coatings; also, even if viscosity is not the key property of interest, it can impact our ability to do closed-loop experimentation. For example, unexpected increases in viscosity can cause liquid-handling robots to fail. Traditional viscosity measurements are manual, invasive, and slow. Here we use convolutional neural networks (CNNs) as an alternative to traditional viscometry by non-invasively extracting the spatiotemporal features of fluid motion under flow. To do this, we built a workflow using a dual-armed collaborative robot that collects video data of fluid motion autonomously. This dataset was then used to train a 3-dimensional convolutional neural network (3D-CNN) for viscosity estimation, either by classification or by regression. We also used these models to identify unknown laboratory solvents, again based on differences in fluid motion. The 3D-CNN model performance was compared with the performance of a panel of human participants for the same classification tasks. Our models strongly outperformed human classification in both cases. For example, even with training on fewer than 50 videos for each liquid, the 3D-CNN model gave an average accuracy of 88% for predicting the identity of five different laboratory solvents, compared to an average accuracy of 32% for human observation. For comparison, random category selection would give an average accuracy of 20%. Our method offers an alternative to traditional viscosity measurements for autonomous chemistry workflows that might be used both for process control (, choosing not to pipette liquids that are too viscous) or for materials discovery (, identifying new polymerization catalysts on the basis of viscosification).
PubMed: 38013903
DOI: 10.1039/d3dd00109a -
Sensors (Basel, Switzerland) Nov 2023A pipette-free and fully integrated device that can be used to accurately recognize the presence of infectious pathogens is an important and useful tool in point-of-care...
Pipette-Free and Fully Integrated Paper Device Employing DNA Extraction, Isothermal Amplification, and Carmoisine-Based Colorimetric Detection for Determining Infectious Pathogens.
A pipette-free and fully integrated device that can be used to accurately recognize the presence of infectious pathogens is an important and useful tool in point-of-care testing, particularly when aiming to decrease the unpredictable threats posed by disease outbreak. In this study, a paper device is developed to integrate the three main processes required for detecting infectious pathogens, including DNA extraction, loop-mediated isothermal amplification (LAMP), and detection. All key reagents, including sodium dodecyl sulfate (SDS), NaOH, LAMP reagents, and carmoisine, are placed on the paper device. The paper device is operated simply via sliding and folding without using any bulky equipment, and the results can be directly observed by the naked eye. The optimized concentrations of sodium dodecyl sulfate (SDS), sodium hydroxide (NaOH), and carmoisine were found to be 0.1%, 0.1 M, and 0.5 mg/mL, respectively. The paper device was used to detect at concentrations as low as 10 CFU/mL within 60 min. Also, spiked in milk was successfully detected using the paper device, demonstrating the feasible application in real sample analysis.
Topics: Colorimetry; Sodium Dodecyl Sulfate; Sodium Hydroxide; Nucleic Acid Amplification Techniques; DNA
PubMed: 38005500
DOI: 10.3390/s23229112