We provide a basis for automated single-cell sorting based on optical trapping and manipulation using human peripheral blood as a model system. A counterpropagating dual-beam optical-trapping configuration is shown theoretically and experimentally to be preferred due to a greater ability to manipulate cells in three dimensions. Theoretical analysis performed by simulating the propagation of rays through the region containing an erythrocyte (red blood cell) divided into numerous elements confirms experimental results showing that a trapped erythrocyte orients with its longest axis in the direction of propagation of the beam. The single-cell sorting system includes an image-processing system using thresholding, background subtraction, and edge-enhancement algorithms, which allows for the identification of single cells. Erythrocytes have been identified and manipulated into designated volumes using the automated dual-beam trap. Potential applications of automated single-cell sorting, including the incorporation of molecular biology techniques, are discussed.

We present a theoretical analysis of the behaviour of erythrocytes in an optical trapping system. We modeled erythrocyte behaviour in an optical trap by an algorithm which divided the cell surface into a large number of elements and recursively summed the force and torque on each element. We present a relationship between the torque and angle of orientation of the cell, showing that stable equilibrium orientations are at angles of 0 o , 180 o and 360 o and unstable equilibrium orientations are at 90 o and 270 o relative to the axis of beam propagation. This is consistent with our experimental observations and with results described in the literature. We also model behaviour of the erythrocyte during micromanipulation by calculating the net force on it. Such theoretical analysis is practical as it allows for the optimization of the optical parameters of a trapping system prior to performing a specific optical micromanipulation application, such as cell sorting or construction of a cell pattern for lab-on-a-chip applications.