RESUMEN
The primary goal of bioprocess cell line development is to obtain high product yields from robustly growing and well-defined clonal cell lines in timelines measured in weeks rather than months. Likewise, high-throughput screening of B cells and hybridomas is required for most cell line engineering workflows. A substantial bottleneck in these processes is detecting and isolating rare clonal cells with the required characteristics. Traditionally, this was achieved by the resource-intensive method of limiting dilution cloning, and more recently aided by semiautomated technologies such as cell sorting (e.g., fluorescence-activated cell sorting) and colony picking. In this paper we report on our novel Cyto-Mine Single Cell Analysis and Monoclonality Assurance System, which overcomes the limitations of current technologies by screening hundreds of thousands of individual cells for secreted target proteins, and then isolating and dispensing the highest producers into microtiter plate wells (MTP). The Cyto-Mine system performs this workflow using a fully integrated, microfluidic Cyto-Cartridge. Critically, all reagents and Cyto-Cartridges used are animal component-free (ACF) and sterile, thus allowing fast, robust, and safe isolation of desired cells.
Asunto(s)
Células Clonales/citología , Ensayos Analíticos de Alto Rendimiento/métodos , Análisis de la Célula Individual/métodos , Programas Informáticos , Animales , Antígenos/metabolismo , Células CHO , Células Inmovilizadas/citología , Cricetulus , Citometría de Flujo , Transferencia Resonante de Energía de Fluorescencia , Humanos , Procesamiento de Imagen Asistido por Computador , Inmunoglobulina G/metabolismo , RatonesRESUMEN
Although CCD X-ray detectors can be faster to use, their large-area versions can be much more expensive than similarly sized photographic plate detectors. When indexing X-ray diffraction patterns, large-area detectors can prove very advantageous as they provide more spots, which makes fitting an orientation easier. On the other hand, when looking for single crystals in a polycrystalline sample, the speed of CCD detectors is more useful. A new setup is described here which overcomes some of the limitations of limited-range CCD detectors to make them more useful for indexing, whilst at the same time making it much quicker to find single crystals within a larger polycrystalline structure. This was done by combining a CCD detector with a six-axis goniometer, allowing the compilation of images from different angles into a wide-angled image. Automated scans along the sample were coupled with image processing techniques to produce grain maps, which can then be used to provide a strategy to extract single crystals from a polycrystal.
Asunto(s)
Anestesiología/normas , Ecocardiografía Transesofágica/normas , Anestesia , Consenso , Embolia Aérea/diagnóstico por imagen , Corazón/fisiología , Cardiopatías Congénitas/cirugía , Enfermedades de las Válvulas Cardíacas/diagnóstico por imagen , Humanos , Procedimientos Neuroquirúrgicos , Atención Perioperativa , Embolia Pulmonar/diagnóstico por imagen , Garantía de la Calidad de Atención de Salud , Sociedades Médicas , Estados Unidos , Función Ventricular Izquierda , Función Ventricular DerechaAsunto(s)
Ecocardiografía Transesofágica/normas , Adulto , Anestesiología , Cardiología/educación , Ecocardiografía , Ecocardiografía Transesofágica/métodos , Embolia Aérea/diagnóstico por imagen , Cardiopatías Congénitas/diagnóstico por imagen , Humanos , Derrame Pericárdico/diagnóstico por imagen , Periodo Perioperatorio , Garantía de la Calidad de Atención de Salud , Sociedades Médicas , Traumatismos Torácicos/diagnóstico por imagen , Estados UnidosAsunto(s)
Válvula Aórtica/diagnóstico por imagen , Atrios Cardíacos/diagnóstico por imagen , Implantación de Prótesis de Válvulas Cardíacas/efectos adversos , Complicaciones Posoperatorias/diagnóstico por imagen , Válvula Aórtica/cirugía , Diagnóstico Precoz , Ecocardiografía Transesofágica , Femenino , Fístula/diagnóstico por imagen , Fístula/etiología , Atrios Cardíacos/cirugía , Humanos , Persona de Mediana Edad , Complicaciones Posoperatorias/etiologíaAsunto(s)
Procedimientos Quirúrgicos Cardíacos/métodos , Ecocardiografía Tridimensional/métodos , Ecocardiografía Transesofágica/métodos , Insuficiencia de la Válvula Mitral/diagnóstico por imagen , Cateterismo Cardíaco , Femenino , Fluoroscopía , Defectos del Tabique Interatrial/diagnóstico por imagen , Defectos del Tabique Interatrial/cirugía , Válvulas Cardíacas/cirugía , Humanos , Persona de Mediana Edad , Insuficiencia de la Válvula Mitral/cirugía , Monitoreo Intraoperatorio , StentsRESUMEN
Stroke and neurological injury are among the most devastating and disabling complications associated with cardiac surgery. Transesophageal echocardiography and epiaortic ultrasound allow for sensitive, point-of-care diagnosis of thoracic aortic disease, which is especially common in patients with heart disease. Unlike other operative procedures, the manipulation of the ascending aorta is routine in cardiac surgery and often unavoidable. Dislodgement and embolization from the ascending and aortic arch atheromas have been clearly associated with manipulation during cardiac surgery. Epiaortic ultrasound and transesophageal echocardiography screening are more accurate and more accessible to the operative team than any other available modality to diagnose atherosclerosis of the aorta. The goal of this review is to review the rationale and scientific evidence that suggests that the routine use of ultrasound guidance in cardiac surgery may improve postoperative outcomes in this patient population.