Abstract
The Fluorescence-Activated Cell Sorter (FACS) is a well-established instrument used for identifying, enumerating, classifying and sorting cells by their physical and optical characteristics. For a miniaturized FACS device, a disposable plastic microchip has been developed which has a hydrodynamic focusing chamber using soft lithography. As the characteristics of the spatially confined sample stream have an effect on sample throughput, detection efficiency, and the accuracy of cell sorting, systematic fluid dynamic studies are required. Flow visualization is conducted with a laser scanning confocal microscopy (LSCM), and three-dimensional flow structure of the focused sample stream is reconstructed from 2D slices acquired at $1\mutextrm{m}$ intervals in depth. It was observed that the flow structure in the focusing chamber is skewed by unsymmetrical velocity profile arising from trapezoidal cross section of the microchannel. For a quantitative analysis of a microscopic flow structure, Confocal Micro-PIV system has been developed to evaluate the accelerated flow field in the focusing chamber. This study proposes a method which defines the depth of the measurement volume using a detection pinhole. The trajectories of red blood cells (RBCs) and their interactions with surrounding flow field in the squeezed sample stream are evaluated to find optimal shape of the focusing chamber and fluid manipulation scheme for stable cell transporting, efficient detection, and sorting