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This protocol demonstrates the separation of living cells with the microfluidic dielectrophoresis chip, using the Jurkat cell as a model. The successful living cell separation lies in familiarity with the detailed tips, which are aided by this stepwise protocol. The knowledge of correct chip installation, sample and buffer filling, flow rate and cell concentration adjustments, and contamination sources increases the efficiency of target viable cell collection. Such instructions, although trivial, are critical for achieving cell separation. For complete details on the use and execution of this protocol, please refer to Oshiro et al. (2022).
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Técnicas Analíticas Microfluídicas , Separación Celular/métodos , Técnicas Analíticas Microfluídicas/métodos , MicrofluídicaRESUMEN
Microfluidic dielectrophoresis (DEP) technology has been applied to many devices to perform label-free target cell separation. Cells separated by these devices are used in laboratories, mainly for medical research. The present study designed a microfluidic DEP device to fabricate a rapid and semiautomated cell separation system in conjunction with microscopy to enumerate the separated cells. With this device, we efficiently segregated bacterial cells from liquid products and enriched one cell type from two mixed eukaryotic cell types. The device eliminated sample pretreatment and established cell separation by all-in-one operation in a lab-on-chip, requiring only a small sample volume (0.5-1 mL) to enumerate the target cells and completing the entire separation process within 30 min. Such a rapid cell separation technique is in high demand by many researchers to promptly characterize the target cells.
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BACKGROUND: PixeeMo™ is a compact instrument that enables bacterial cell counting using microfluidic chips instead of counting of colonies on culture media. Chips containing electrodes, based on fluid, electric filtering and sorting technology (FES), allow the selection of bacterial cells from other components in the sample. In the United States (US), surface water or ground water affected by surface water must be treated to reduce the total microbial load to less than 500 CFU/mL. In Japan, drinking water regulations limit the total bacterial load to 100 CFU/mL. OBJECTIVE: To validate the PixeeMoTM aerobic bacteria method based on the Japanese regulation in the range of 30-300 CFU/mL in drinking water. METHOD: PixeeMoTM aerobic bacteria method was compared to the Standard Method for the Examination of Water and Wastewater (SMEWW) 9215B (2017) using naturally contaminated drinking water. RESULTS: The maximum repeatability standard deviation of the PixeeMoTM method was 14.8%. The difference of mean log10 values between the PixeeMoTM and SMEWW 9215B methods ranged from -0.015 to 0.258. Similar results were obtained in the independent laboratory study. CONCLUSIONS: The PixeeMoTM method is equivalent to that of the SMEWW 9215B methods. The product consistency and stability study demonstrated no significant difference within the expiration date. The robustness study confirmed that there was no effect within the expected range. The instrument variation study also demonstrated no significant difference among the data of three PixeeMoTM instruments. HIGHLIGHTS: Total counts of bacteria in drinking water can be determined accurately within 1 h with PixeeMoTM.
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Bacterias Aerobias , Agua Potable , Bacterias , Carga Bacteriana , Recuento de Colonia Microbiana , JapónRESUMEN
OBJECTIVES: The aim of this study is to clarify factors that support the work engagement of nurses, who bear the burden of extended day shifts, by focusing on the advantages of the variable shift system and workday break activities. METHODS: Nurses who were working under a variable shift system were asked to complete a self-report questionnaire to examine the workload, work engagement, work stressors, stress-coping strategies, and stress-coping break time activities, as well as the advantages and disadvantages of the variable shift system. Nine break activities were classified into the following four categories: social activities, rest/relaxation, entertainment, and cognitive activities. The advantages or disadvantages of the variable shift system were scored by developing composite variables using principal component analysis. These variables were used to perform a multiple regression analysis with work engagement as the dependent variable. RESULTS: The advantage score was the variable most strongly correlated with work engagement. In contrast, "Quantitative workload" was negatively correlated with work engagement. Among break activities, in the social activities category correlations were observed in "Both conversation and Email/SNS" and "Conversation only". Although in fact most nurses chose conversation as one of the break options, more than half of the nurses selected rest/relaxation as their ideal break activity. CONCLUSION: Our study suggested that the variable shift system supported the work engagement of nurses who worked extended day shifts. The results also suggested that it would be useful to arrange the employee lounge environment so that employees could freely choose between "conversation" or "taking a rest" depending on the circumstances.