RESUMO
Autonomous migration is essential for the function of immune cells such as neutrophils and plays an important role in numerous diseases. The ability to routinely measure or target it would offer a wealth of clinical applications. Video microscopy of live cells is ideal for migration analysis, but cannot be performed at sufficiently high-throughput (HT). Here we introduce ComplexEye, an array microscope with 16 independent aberration-corrected glass lenses spaced at the pitch of a 96-well plate to produce high-resolution movies of migrating cells. With the system, we enable HT migration analysis of immune cells in 96- and 384-well plates with very energy-efficient performance. We demonstrate that the system can measure multiple clinical samples simultaneously. Furthermore, we screen 1000 compounds and identify 17 modifiers of migration in human neutrophils in just 4 days, a task that requires 60-times longer with a conventional video microscope. ComplexEye thus opens the field of phenotypic HT migration screens and enables routine migration analysis for the clinical setting.
Assuntos
Cristalino , Lentes , Humanos , Microscopia , Microscopia de Vídeo , Movimento CelularRESUMO
Light-sheet fluorescence microscopy (LSFM) can produce high-resolution tomograms of tissue vasculature with high accuracy. However, data processing and analysis is laborious due to the size of the datasets. Here, we introduce VesselExpress, an automated software that reliably analyzes six characteristic vascular network parameters including vessel diameter in LSFM data on average computing hardware. VesselExpress is â¼100 times faster than other existing vessel analysis tools, requires no user interaction, and integrates batch processing and parallelization. Employing an innovative dual Frangi filter approach, we show that obesity induces a large-scale modulation of brain vasculature in mice and that seven other major organs differ strongly in their 3D vascular makeup. Hence, VesselExpress transforms LSFM from an observational to an analytical working tool.
Assuntos
Imageamento Tridimensional , Software , Animais , Camundongos , Imageamento Tridimensional/métodos , Microscopia de Fluorescência/métodos , Encéfalo/diagnóstico por imagemRESUMO
OBJECTIVE: The aim of this study was to determine the prevalence and the characteristics of coronavirus disease 2019 (COVID-19) in a tertiary outpatient clinic of asthma patients, find the predisposing asthma phenotype to COVID-19, and to see their adherence to asthma treatment. MATERIAL AND METHODS: A retrospective, cross-sectional, real life study was conducted via phone interviews with the patients being followed in the asthma outpatient clinic. From the files of the patient information was obtained about their demographics, asthma phenotype, co-morbidity, prick tests, spirometry test results and their medications at the last visit before the COVID-19 pandemic. Information's about asthma exacerbations, ACT, asthma treatment adherence and history of COVID-19 were obtained via telephone interviews. RESULTS: Of the 573 patients with asthma, 13 (2.26%) had COVID-19 history. The mean age of patients with asthma and COVID-19 was 51.84±14.92 year. Two patients were on mepolizumab and one was on omalizumab treatment. Mean ACT was 19.84±2.73. Lack of adherence was reported in 8% of all patients with asthma compared to 23% in the patients who had COVID-19. Asthma exacerbation was seen during the course of SARS-CoV2 infection in 3 of 13 patients with asthma. Asthma exacerbations were reported during the period of one month following COVID-19 in 2 patients. CONCLUSION: The most common asthma phenotype in the cases of COVID-19 was obese phenotype. Rates of using biological agents and non-adherence to the treatment were found to be higher. Asthma exacerbation may be seen during course of COVID-19 albeit being less common.