Current robots can manipulate only surface-attached cells seriously limiting the fields of their application for single cell handling. We developed a computer vision-based robot applying a motorized microscope and micropipette to recognize and gently isolate intact individual cells for subsequent analysis, e.g., DNA/RNA sequencing in 1–2 nanoliters from a thin (~100 μm) layer of cell suspension. It can retrieve rare cells, needs minimal sample preparation, and can be applied for virtually any tissue cell type. Combination of 1 μm positioning precision, adaptive cell targeting and below 1 nl liquid handling precision resulted in an unprecedented accuracy and efficiency in robotic single cell isolation. Single cells were injected either into the wells of a miniature plate with a sorting speed of 3 cells/min or into standard PCR tubes with 2 cells/min. We could isolate labeled cells also from dense cultures containing ~1,000 times more unlabeled cells by the successive application of the sorting process. We compared the efficiency of our method to that of single cell entrapment in microwells and subsequent sorting with the automated micropipette: the recovery rate of single cells was greatly improved.
This paper presents and compares two different strategies in the numerical simulation of passive microfluidic mixers based on chaotic advection. In addition to flow velocity field calculations, concentration distributions of molecules and trajectories of microscale particles were determined and compared to evaluate the performance of the applied modeling approaches in the proposed geometries. A staggered herringbone type micromixer (SHM) was selected and studied in order to demonstrate finite element modeling issues. The selected microstructures were fabricated by a soft lithography technique, utilizing multilayer SU-8 epoxy-based photoresist as a molding replica for polydimethylsiloxane (PDMS) casting. The mixing processes in the microfluidic systems were characterized by applying molecular and particle (cell) solutions and adequate microscopic visualization techniques. We proved that modeling of the molecular concentration field is more costly, in regards to computational time, than the particle trajectory based method. However, both approaches showed adequate qualitative agreement with the experimental results.
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