RESUMO
A low-cost, handheld centrifugal microfluidic system for multiplexed visual detection based on recombinase polymerase amplification (RPA) was developed. A concise centrifugal microfluidic chip featuring four reaction units was developed to run multiplexed RPA amplification in parallel. Additionally, a significantly shrunk-size and cost-effective handheld companion device was developed, incorporating heating, optical, rotation, and sensing modules, to perform multiplexed amplification and visual detection. After one-time sample loading, the metered sample was equally distributed into four separate reactors with high-speed centrifugation. Non-contact heating was adopted for isothermal amplification. A tiny DC motor on top of the chip was used to drive steel beads inside reactors for active mixing. Another small DC motor, which was controlled by an elaborate locking strategy based on magnetic sensing, was adopted for centrifugation and positioning. Visual fluorescence detection was optimized from different sides, including material, surface properties, excitation light, and optical filters. With fluorescence intensity-based visual detection, the detection results could be directly observed through the eyes or with a smartphone. As a proof of concept, the handheld device could detect multiple targets, e.g., different genes of African swine fever virus (ASFV) with the comparable LOD (limit of detection) of 75 copies/test compared to the tube-based RPA.
Assuntos
Técnicas de Amplificação de Ácido Nucleico , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Técnicas de Amplificação de Ácido Nucleico/métodos , Vírus da Febre Suína Africana/isolamento & purificação , Vírus da Febre Suína Africana/genética , Dispositivos Lab-On-A-Chip , Limite de Detecção , Centrifugação/instrumentação , Animais , Smartphone , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/economiaRESUMO
To improve the accuracy of modal analysis for a four-stage centrifugal-pump rotor system with a balancing disc based on the concentrated-mass analytical method, a simplified concentrated mass mathematical model and an ANSYS simulation model are established. The results from these two models are compared to determine factors that cause significant differences in the mode shapes. Subsequently, an optimized mathematical model based on the corrected mass moment of an inertia matrix and stiffness correction coefficients is proposed, and the effectiveness of this optimized mathematical model is validated using a four-stage centrifugal pump with back blades. The results show that the natural frequencies obtained from the ANSYS simulations are consistently higher than those obtained using the analytical method. The simplification of the moment of inertia at the impeller and balancing disc contributes primarily to the calculated errors. The optimized mathematical model reduces the errors in the natural frequencies from 12.96%, 12.13%, 9.96%, 5.85%, and 8.74% to 2.45%, 1.56%, 0.65%, 5.34%, and 2.28%, respectively. The optimization of natural frequencies offers better performance at lower-order modes, whereas its effects on higher-order modes are less significant. The optimization method is applicable to centrifugal pumps with back blades and reduces the error in theoretical calculations, based on reductions in the concentrated mass from 13.11%, 12.85%, 9.91%, and 7.2% to 3.7%, 3.86%, 0.57%, and 2.87%, respectively, thus further confirming the feasibility of the optimized model design.
Assuntos
Centrifugação , Modelos Teóricos , Centrifugação/instrumentação , Centrifugação/métodos , Simulação por Computador , Desenho de EquipamentoRESUMO
Point of care testing (POCT) of nucleic acids holds significant importance in the realm of infectious disease prevention and control, as well as the advancement of personalized precision medicine. Nevertheless, conventional nucleic acid testing methods continue to face challenges such as prolonged detection times and dependence on extensive specialized equipment and personnel, rendering them unsuitable for point of care applications. Here, we proposed an innovative active centrifugal microfluidic system (ACMS) for automatic nucleic acid extraction, encompassing modules for active valve control and magnetic control. An on-chip centrifugal puncture valve (PV) was devised based on the elastic tolerance differences between silicone membranes and tinfoils to release pre-embedded liquid reagents on demand. Furthermore, we have utilized the returnable valve (RV) technology to accurately control the retention and release of liquids, leveraging the high elastic tolerance of the silicone membrane. By incorporating an online controllable magnetic valve, we have achieved controlled and rapid aggregation and dispersion of magnetic beads. The final chip encapsulates multiple reagents and magnetic beads necessary for nucleic acid extraction. Upon sample addition and loading into the instrument, automated on-chip sample loading and nucleic acid extraction, purification, and collection can be accomplished within 30 minutes, halving the overall operation time and even increasing the efficiency of pseudovirus extraction by three orders of magnitude. Consequently, real-time fluorescence quantitative PCR amplification has successfully detected multiple targets of the SARS-CoV-2 virus (with an impressive detection limit as low as 10 copies per µL), along with targeted sequencing analysis yielding a conformity rate of 99%.
Assuntos
Centrifugação , Dispositivos Lab-On-A-Chip , Centrifugação/instrumentação , Humanos , SARS-CoV-2/isolamento & purificação , SARS-CoV-2/genética , Técnicas Analíticas Microfluídicas/instrumentação , Desenho de Equipamento , Ácidos Nucleicos/isolamento & purificação , Ácidos Nucleicos/análise , RNA Viral/isolamento & purificação , RNA Viral/análise , COVID-19/diagnóstico , COVID-19/virologiaRESUMO
Immunoassays serve as powerful diagnostic tools for early disease screening, process monitoring, and precision treatment. However, the current methods are limited by high costs, prolonged processing times (>2 h), and operational complexities that hinder their widespread application in point-of-care testing. Here, we propose a novel centrifugo-pneumatic reciprocating flowing coupled with spatial confinement strategy, termed PRCM, for ultrafast multiplexed immunoassay of pathogens on a centrifugal microfluidic platform. Each chip consists of four replicated units; each unit allows simultaneous detection of three targets, thereby facilitating high-throughput parallel analysis of multiple targets. The PRCM platform enables sequential execution of critical steps such as solution mixing, reaction, and drainage by coordinating inherent parameters, including motor rotation speed, rotation direction, and acceleration/deceleration. By integrating centrifugal-mediated pneumatic reciprocating flow with spatial confinement strategies, we significantly reduce the duration of immune binding from 30 to 5 min, enabling completion of the entire testing process within 20 min. As proof of concept, we conducted a simultaneous comparative test on- and off-the-microfluidics using 12 negative and positive clinical samples. The outcomes yielded 100% accuracy in detecting the presence or absence of the SARS-CoV-2 virus, thus highlighting the potential of our PRCM system for multiplexed point-of-care immunoassays.
Assuntos
COVID-19 , Centrifugação , SARS-CoV-2 , Imunoensaio/métodos , Imunoensaio/instrumentação , SARS-CoV-2/isolamento & purificação , Centrifugação/instrumentação , COVID-19/diagnóstico , COVID-19/virologia , Humanos , Técnicas Analíticas Microfluídicas/instrumentação , Dispositivos Lab-On-A-ChipRESUMO
Cytocentrifugation is a common technique for the capture of cells on microscopic slides. It usually requires a special cytocentrifuge or cytorotor and cassettes. In the study presented here, we tested the new concept of cytocentrifugation based on the threaded connection of the lid and the sample holder to ensure an adjustable flow of solutions through the filters and the collection of the filtered solutions in the reservoir during centrifugation. To test this concept, we developed a device for the preparation of cell samples on circular coverslips. The device was tested for the capture and sample processing of both eukaryotic and prokaryotic cells, cell nuclei, and mitochondria for microscopy analysis including image cytometry. Moreover, an efficient procedure was developed for capturing formaldehyde-fixed cells on non-treated coverslips without cell drying. The results showed that the tested arrangement enables the effective capture and processing of all of the tested samples and the developed device represents an inexpensive alternative to common cytocentrifuges, as only the paper filter is consumed during sample processing, and no special centrifuge, cytorotor, or cassette is necessary. As no additional system of solution removal is required during sample staining, the tested concept also facilitates the eventual automation of the staining procedure.
Assuntos
Centrifugação/instrumentação , Técnicas Citológicas/instrumentação , Técnicas Citológicas/métodos , Animais , Centrifugação/métodos , Humanos , Microscopia/métodos , Manejo de Espécimes/métodos , Coloração e Rotulagem/métodosRESUMO
Leukocytes have an essential role in patient clinical trajectories and progression. Traditional methods of leukocyte enrichment have many significant limitations for current applications. It is demonstrated a novel 3D printing leukocyte sorting accumulator that combines with centrifugation to ensure label-free initial leukocyte enrichment based on cell density and size. The internal structure of leukocyte sorting accumulator (revealed here in a new design, leukocyte sorting accumulator-3, upgraded from earlier models), optimizes localization of the buffy coat fraction and the length of the period allocated for a second centrifugation step to deliver a higher recovery of buffy coats than earlier models. Established methodological parameters were evaluated for reliability by calculating leukocyte recovery rates and erythrocyte depletion rates by both pushing and pulling methods of cell displacement. Results indicate that leukocyte sorting accumulator-3 achieves a mean leukocytes recovery fraction of 96.2 ± 2.38% by the pushing method of layer displacement. By the pulling method, the leukocyte sorting accumulator-3 yield a mean leukocytes recovery fraction of 94.4 ± 0.8%. New procedures for preliminary enrichment of leukocytes from peripheral blood that avoid cellular damage, as well as avert metabolic and phase cycle intervention, are required as the first step in many modern clinical and basic research assays.
Assuntos
Procedimentos de Redução de Leucócitos/métodos , Leucócitos/citologia , Impressão Tridimensional/instrumentação , Buffy Coat/classificação , Buffy Coat/citologia , Centrifugação/instrumentação , Centrifugação/métodos , Humanos , Procedimentos de Redução de Leucócitos/instrumentação , Leucócitos/classificaçãoRESUMO
BACKGROUND: We present the analytical performance of the ivisen IA-1400, a new point-of-care device that features a characteristic built-in centrifuge system, to measure blood cardiac troponin I (cTnI) levels. METHODS: Whole blood and plasma samples obtained from patients who visited Korea University Guro Hospital were used to analyze measurement range, cross-reactivity, interference, and sensitivity and specificity. We performed a correlation analysis of the ivisen IA-1400 versus the Access AccuTnI+3 immunoassay using the UniCel™ DxI 800 platform and the PATHFAST™ hs-cTnI assay. RESULTS: Within-run precisions were classified as low, 9.8%; middle, 10.2%; and high, 8.5%. The limit of blank was 3.1 ng/L for plasma samples and 4.3 ng/L for whole blood samples. The limit of detection was 8.4 ng/L for plasma samples and 10.0 ng/L for whole blood samples, respectively. The limit of quantitation at a coefficient of variation of 20% and 10% was 19.5 ng/L and 45.5 ng/L for plasma samples, respectively. The comparative evaluation between the two other assays and ivisen IA-1400 showed excellent correlation, with Spearman's correlation coefficients (R) of 0.992 and 0.985. The sensitivity and specificity of ivisen IA-1400 using the optimum cut-off value of 235 ug/L were 94.6% and 98.2%, respectively. CONCLUSION: The ivisen IA-1400 showed acceptable and promising performance in cTnI measurements using whole blood and plasma samples, with limited information in the clinical performance. The flexibility for sample selection using the internal centrifugation system is the main advantage of this point-of-care device.
Assuntos
Centrifugação/instrumentação , Miocárdio/metabolismo , Sistemas Automatizados de Assistência Junto ao Leito , Troponina I/sangue , Intervalos de Confiança , Reações Cruzadas , Humanos , Limite de Detecção , Sensibilidade e EspecificidadeRESUMO
Adulteration of food ingredients, particularly replacement of high-value milk with low-cost milk, affects food safety. For rapid and accurate identification of the possible adulterating milk species in an unknown sample, a centrifugal microfluidic chip-based real-time fluorescent multiplex loop-mediated isothermal amplification (LAMP) assay was developed to simultaneously detect milk from cow, camel, horse, goat, and yak. Using precoated primers in different reaction wells, the centrifugal microfluidic chip markedly simplified the detection process and reduced false-positive results. The entire amplification was completed within 90 min with a genomic detection limit of 0.05 ng/µL in cow, camel, horse, and goat milk and 0.005 ng/µL in yak milk. Using simulated adulterated samples for validation, the detection limit for adulterated milk samples was 2.5%, satisfying authentication requirements, as the proportion of adulterated milk higher than 10% affects economic interests. Therefore, this simple, centrifugal, microfluidic chip-based multiplex real-time fluorescent LAMP assay can simultaneously detect common milk species in commercial products to enable accurate labeling.
Assuntos
Centrifugação/instrumentação , Qualidade dos Alimentos , Dispositivos Lab-On-A-Chip , Leite/química , Técnicas de Diagnóstico Molecular/instrumentação , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Animais , Bovinos , Primers do DNA/genética , Feminino , Leite/normas , Fatores de TempoRESUMO
Introduction: Echinoderm coelomocytes have traditionally been investigated through a morphological approach using light microscopy, which relies on the idea of constant cell shape as a stable character. However, this can be affected by biotic or abiotic conditions. Objective: To analyze if the consistency in cell morphology offered by the cytocentrifugation method, might be used as a convenient tool to study echinoderm coelomocytes. Methods: Cells of Echinaster (Othilia) brasiliensis (Asteroidea), Holothuria (Holothuria) tubulosa (Holothuroidea), Eucidaris tribuloides, Arbacia lixula, Lytechinus variegatus, and Echinometra lucunter (Echinoidea) were spread on microscope slides by cytocentrifugation, stained, and analyzed through light microscopy. Additionally, fluorescence microscopy, scanning electron microscopy, and energy-dispersive x-ray spectroscopy were applied to cytospin preparations, to complement the analysis of granular and colorless spherulocytes of Eucidaris tribuloides. Results: Altogether, 11 cell types, including phagocytes, spherulocytes, vibratile cells, and progenitor cells were identified in the samples analyzed. The granular spherulocyte, a newly-described cell type, was observed in all Echinoidea and was very similar to the acidophilic spherulocytes of Holothuria (Holothuria) tubulosa. Conclusions: Cytocentrifugation proved to be versatile, either as the main method of investigation in stained preparations, or as a framework on which other procedures may be performed. Its ability to maintain a constant morphology allowed accurate correspondence between live and fixed/stained cells, differentiation among similar spherulocytes as well as comparisons between similar cells of Holothuroidea and Echinoidea.
Introducción: Los celomocitos de equinodermos se han investigado tradicionalmente a través de un enfoque morfológico utilizando microscopía óptica, que se basa en la idea de la forma celular constante como un carácter estable. Sin embargo, esto puede verse afectado por condiciones bióticas o abióticas. Objetivo: Analizar si la consistencia en la morfología celular que ofrece el método de citocentrifugación podría utilizarse como una herramienta conveniente para estudiar los celomocitos de equinodermos. Métodos: Células de Echinaster (Othilia) brasiliensis (Asteroidea), Holothuria (Holothuria) tubulosa (Holothuroidea), Eucidaris tribuloides, Arbacia lixula, Lytechinus variegatus y Echinometra lucunter (Echinoidea) se esparcieron en portaobjetos de microscopio por citocentrifugación, se tiñeron y analizaron mediante microscopía óptica. Adicionalmente, se aplicó microscopía de fluorescencia, microscopía electrónica de barrido y espectroscopía de rayos X con dispersión de energía a las preparaciones de citoespina, para complementar el análisis de los esferulocitos granulares e incoloros de Eucidaris tribuloides. Resultados: En total, se identificaron en las muestras analizadas 11 tipos de células, incluidos fagocitos, esferulocitos, células vibrátiles y células progenitoras. El esferulocito granular, un tipo de célula recién descrito, se observó en todos los Echinoidea y fue muy similar a los esferulocitos acidófilos de Holothuria (Holothuria) tubulosa. Conclusiones: La citocentrifugación demostró ser un método bastante versátil, ya sea como el método principal de investigación en preparaciones teñidas o como un marco en el que se pueden realizar otros procedimientos. Su capacidad para mantener una morfología constante permitió una correspondencia precisa entre las células vivas y las células fijas/teñidas, la diferenciación entre esferulocitos similares, así como comparaciones entre células similares de Holothuroidea y Echinoidea.
Assuntos
Animais , Espectrometria por Raios X/métodos , Equinodermos/microbiologia , Centrifugação/instrumentação , Forma do Núcleo CelularRESUMO
BACKGROUND: Cryoprecipitate (CRYO) is neither produced nor supplied by the Japanese Red Cross Society. A novel CRYO extraction method established in-house by modifying a thaw-siphon technique was demonstrated in this study. STUDY DESIGN AND METHODS: A pack of fresh frozen plasma was thawed and equally divided into two bags for CRYO extraction by different methods. CRYO was extracted from the blood plasma using a standard centrifugation method and our modified thaw-siphon method (Bokutoh-siphon method; B method). The two different CRYOs extracted were analyzed to compare the differences in the amount of fibrinogen recovered, clotting factors extracted, and clotting activity. RESULTS: The amount of fibrinogen in the CRYO extracted using the B-siphon method was similar to that obtained using the standard method (recovery of fibrinogen: B-siphon method: 71.2% vs. standard method: 61.0%). The amount of clotting XIII factor extracted using the B-siphon method was significantly lower than those extracted using the standard method. On the other hand, clotting II, V factors, and C1q esterase inhibitor not concentrated in CRYO content from the B-siphon method were significantly higher than that from the standard method. CONCLUSION: A new in-house CRYO preparation method was established by modifying a previously used thaw-siphon method. A coagulation factor-rich CRYO was extracted from plasma frozen at -40°C along with the first fraction of thawed plasma, without using a large-capacity refrigerated centrifuge for blood bags.
Assuntos
Fatores de Coagulação Sanguínea/análise , Centrifugação/instrumentação , Criopreservação/métodos , Fibrinogênio/análise , Plasma/química , Fatores de Coagulação Sanguínea/metabolismo , Precipitação Química , Proteína Inibidora do Complemento C1/metabolismo , Fator V/análise , Fator VIII/análise , Fibrinogênio/metabolismo , Humanos , Indicadores e Reagentes/química , Protrombina/análiseRESUMO
Infection caused by the new coronavirus (SARS-CoV-2) has become a serious worldwide public health problem, and one of the most important strategies for its control is mass testing. Loop-mediated isothermal amplification (LAMP) has emerged as an important alternative to simplify the diagnostics of infectious diseases. In addition, an advantage of LAMP is that it allows for easy reading of the final result through visual detection. However, this step must be performed with caution to avoid contamination and false-positive results. LAMP performed on microfluidic platforms can minimize false-positive results, in addition to having potential for point-of-care applications. Here, we describe a polystyrene-toner (PS-T) centrifugal microfluidic device manually controlled by a fidget spinner for molecular diagnosis of COVID-19 by RT-LAMP, with integrated and automated colorimetric detection. The amplification was carried out in a microchamber with 5 µL capacity, and the reaction was thermally controlled with a thermoblock at 72 °C for 10 min. At the end of the incubation time, the detection of amplified RT-LAMP fragments was performed directly on the chip by automated visual detection. Our results demonstrate that it is possible to detect COVID-19 in reactions initiated with approximately 10-3 copies of SARS-CoV-2 RNA. Clinical samples were tested using our RT-LAMP protocol as well as by conventional RT-qPCR, demonstrating comparable performance to the CDC SARS-CoV-2 RT-qPCR assay. The methodology described in this study represents a simple, rapid, and accurate method for rapid molecular diagnostics of COVID-19 in a disposable microdevice, ideal for point-of-care testing (POCT) systems.
Assuntos
Teste de Ácido Nucleico para COVID-19/métodos , Determinação de Ponto Final/métodos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Poliestirenos , SARS-CoV-2/isolamento & purificação , Animais , COVID-19/diagnóstico , COVID-19/genética , Teste de Ácido Nucleico para COVID-19/instrumentação , Centrifugação/instrumentação , Centrifugação/métodos , Chlorocebus aethiops , Determinação de Ponto Final/instrumentação , Humanos , Técnicas de Diagnóstico Molecular/instrumentação , Técnicas de Amplificação de Ácido Nucleico/instrumentação , SARS-CoV-2/genética , Fatores de Tempo , Células VeroRESUMO
This chapter covers the various methods of mechanical cell disruption and tissue homogenization that are currently commercially available for processing small samples s < 1 mL) to larger multikilogram production quantities. These mechanical methods of lysing do not introduce chemicals or enzymes to the system. However, the energies required when using these "harsh," high mechanical energy methods can be enough to damage the very components being sought.The destruction of cell membranes and walls is effected by subjecting the cells (a) to shearing by liquid flow, (b) to exploding by pressure differences between inside and outside of cell, (c) to collision forces by impact of beads or paddles, or (d) a combination of these forces.Practical suggestions to optimize each method, where to acquire such equipment, and links to reference sources are included. Several novel technologies are presented.
Assuntos
Fracionamento Celular/instrumentação , Extratos de Tecidos , Animais , Extratos Celulares , Centrifugação/instrumentação , Desenho de Equipamento , Humanos , Pressão , Sonicação/instrumentação , Estresse MecânicoRESUMO
This study evaluated the impact of rotor angle and time of storage after centrifugation on the in vitro biological properties of platelet-rich fibrin (PRF) membranes. Blood samples (n = 9) were processed with a vertical fixed-angle (V) or a swing-out horizontal (H) centrifuge, with 20-60 min of sample storage after centrifugation. Leukocytes, platelets, and red blood cells were counted, and fibrin architecture was observed by scanning electron microscopy (SEM). The release of FGF2, PDGFbb, VEGF, IL-6, and IL-1ß was measured after incubation on culture media for 7-21 days. Cell content was equivalent in all experimental groups (p > .05). The fibrin matrix was similar for fixed-angle and horizontal centrifugation. Horizontal centrifugation induced a twofold increase in PDGF and 1.7× increase on FGF release as compared to V samples, while IL-1ß was significantly reduced (p < .05). No significant difference was observed on the release of growth factors and cytokines at different times after centrifugation (p < .05). These data suggest that both angles of centrifugation produce PRF membranes with similar structure and cellularity, but horizontal centrifugation induces a higher release of growth factors. Higher times of storage after centrifugation did not impact on cell content and the release of growth factors.
Assuntos
Centrifugação/instrumentação , Centrifugação/métodos , Fibrina Rica em Plaquetas/química , Adulto , Plaquetas/química , Citocinas/química , Eritrócitos , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/química , Leucócitos/química , Masculino , Pessoa de Meia-IdadeRESUMO
Artificial gravity elicited through short-arm human centrifugation combined with physical exercise, such as jumping, is promising in maintaining health and performance during space travel. However, motion sickness symptoms could limit the tolerability of the approach. Therefore, we determined the feasibility and tolerability, particularly occurrence of motion sickness symptoms, during reactive jumping exercises on a short-arm centrifuge. In 15 healthy men, we assessed motion sickness induced by jumping exercises during short-arm centrifugation at constant +1Gz or randomized variable +0.5, +0.75, +1, +1.25 and +1.5 Gz along the body axis referenced to center of mass. Jumping in the upright position served as control intervention. Test sessions were conducted on separate days in a randomized and cross-over fashion. All participants tolerated jumping exercises against terrestrial gravity and on the short-arm centrifuge during 1 Gz or variable Gz at the center of mass without disabling motion sickness symptoms. While head movements markedly differed, motion sickness scores were only modestly increased with jumping on the short-arm centrifuge compared with vertical jumps. Our study demonstrates that repetitive jumping exercises are feasible and tolerable during short-arm centrifugation. Since jumping exercises maintain muscle and bone mass, our study enables further development of exercise countermeasures in artificial gravity.
Assuntos
Centrifugação/efeitos adversos , Gravidade Alterada/efeitos adversos , Enjoo devido ao Movimento/etiologia , Voo Espacial , Adaptação Fisiológica , Adulto , Medicina Aeroespacial , Centrifugação/instrumentação , Exercício Físico/fisiologia , Gravitação , Movimentos da Cabeça/fisiologia , Voluntários Saudáveis , Humanos , Masculino , Enjoo devido ao Movimento/fisiopatologia , Enjoo devido ao Movimento/prevenção & controle , Contramedidas de Ausência de Peso , Adulto JovemRESUMO
Implantable ventricular assist devices are a type of mechanical circulatory support for assisting the pumping of the heart. Accurate estimation of the flow rate through such devices is critical to ensure effective performance. A novel method for estimating the flow rate using the passively stabilized position of a magnetically levitated impeller was developed by our group. However, the performance of the method is affected by the gravity vector, which depends on the patient's posture. In this study, the effects of gravity on the flow estimation method are analyzed, and a compensation method is proposed. The magnetically levitated impeller is axially suspended and radially restricted by passive stability in a centrifugal blood pump developed by our group. The gravity effects were evaluated by analyzing the relationships between the radial position of the magnetically levitated impeller and the flow rate with respect to the gravity direction. Accurate estimation of the flow rate could not be achieved when the direction of gravity with respect to impeller was unknown. A mean absolute error of up to 4.89 L/min was obtained for flow rate measurement range of 0-5 L/min. However, analysis of the equilibrium of forces on the passively stabilized impeller indicated that the effects of gravity on the flow estimation could be compensated by performing additional measurements of the gravity direction with respect to impeller. The method for compensating the effects of gravity on the flow estimation should improve the performance of therapy with the implantable ventricular assist devices.
Assuntos
Circulação Assistida/instrumentação , Coração Auxiliar/normas , Hidrodinâmica , Posicionamento do Paciente/métodos , Centrifugação/instrumentação , Desenho de Equipamento , Gravitação , Humanos , Fenômenos MagnéticosRESUMO
INTRODUCTION: The miniaturization of blood pumps has become a trend due to the advantage of easier transplantation, especially for pediatric patients. In small-scale pumps, it is much easier and more cost-efficient to manufacture the impeller with straight blades compared to spiral-profile blades. METHODS: Straight-blade impeller designs with different blade angles, blade numbers, and impeller flow passage positions are evaluated using the computational fluid dynamics method. Blade angles (θ = 0°, 20°, 30°, and 40°), blade numbers (N = 5, 6, 7, and 8), and three positions of impeller flow passage (referred to as top, middle, and bottom) are selected as the studied parametric values. RESULTS: The numerical results reveal that with increasing blade angle, the pressure head and the hydraulic efficiency increase, and the average scalar shear stress and the normalized index of hemolysis decrease. The minimum radial force and axial thrust are obtained when θ equals 20°. In addition, the minimum average scalar shear stress and normalized index of hemolysis values are obtained when N = 6, and the maximum values are obtained when N = 5. Regarding the impeller flow passage position, the axial thrust and the stagnation area forming in the impeller eye are reduced as the flow passage height declines. CONCLUSION: The consideration of a blade angle can greatly improve the performance of blood pumps, although the influence of the blade number is not very easily determined. The bottom position of the impeller flow passage is the best design.
Assuntos
Circulação Assistida/instrumentação , Coração Auxiliar/efeitos adversos , Hemólise , Hidrodinâmica , Centrifugação/instrumentação , Desenho Assistido por Computador , Desenho de Equipamento , Humanos , Miniaturização/métodos , Estresse MecânicoRESUMO
We investigated whether an ordinary centrifuge can achieve the standard centrifugal effect required according to specifications for infectious disease screening using the Abbott i2000. Samples were collected and centrifuged following a standard operating procedure (SOP). They were then divided into three groups according to the results of the initial screening tests: a negative group, weak-positive group, and positive group. Twenty negative samples and all weak-positive and positive samples were re-analyzed. Two tubes for each re-analyzed sample were centrifuged simultaneously, one for 10 min at 10 000 × g, per recommendations, and one for 10 min at 2750 × g. No significant difference was found between the groups using different centrifugal forces. There was a strong correlation in the quantitative values between the two conditions of centrifugation. Consistency analysis showed a Cronbach's alpha > 0.8 for detection of Treponema pallidum, human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B surface antigen in the three groups (negative group, weak-positive group, and positive group) under different centrifugation conditions. Strong consistency was found under different centrifugal conditions, regardless of the initial testing results. In conclusion, we conducted centrifugation steps in duplicate, according to infectious disease screening protocols. Our study showed that all samples should be centrifuged using a recommended relative centrifugal force after a proper clotting time, as in the standard operating procedure of our laboratory. In this way, we were able to obtain the same results using an ordinary centrifuge as those obtained using a high-speed centrifuge, such as the Abbott i2000.
Assuntos
Centrifugação/métodos , Doenças Transmissíveis/diagnóstico , Manejo de Espécimes/instrumentação , Centrifugação/instrumentação , Centrifugação/normas , Guias como Assunto , HIV/isolamento & purificação , Hepacivirus/isolamento & purificação , Antígenos de Superfície da Hepatite B/metabolismo , Vírus da Hepatite B/isolamento & purificação , Humanos , Sensibilidade e Especificidade , Treponema pallidum/isolamento & purificaçãoRESUMO
The tendency of steroid molecules to adsorb to various materials, particularly plastics, has been known of for decades but has not received widespread attention in the scientific community, and a modern, systematic study is lacking. This adsorption is an important consideration for researchers working with steroid hormones as it could skew the results of various experiments. Here we show that steroids adsorb to various vessels used in experiments, including microcentrifuge tubes, glass vials, and cell culture plates, in a manner that depends on the steroid's molecular structure and on the type of vessel. The lipophilicity of steroids is a strong predictor of the degree of adsorption, with nearly 50 % of the most lipophilic steroid tested, pregnenolone, retained in a high-adsorbing microcentrifuge tube after one hour incubation of an aqueous pregnenolone solution followed by removal of the aqueous solvent. We also show the effects of other factors such as incubation time, centrifugation, and temperature on adsorption, and show that adsorption can be mostly prevented by the presence of serum proteins in steroid solutions and/or by the use of low-adsorbing tubes.