An apparatus and method for separating single cells. The apparatus includes: a fluid channel having an upper panel, a lower panel, and a flow path formed therebetween that is configured to convey a sample including a single cell; a single cell measuring unit including first and second electrodes provided on the fluid channel in a predetermined spaced relationship for applying electrical signals to the sample in the flow path, and a detection electrode provided between the first and second electrodes to detect the single cell in the sample in the flow path, such that the single cell measuring unit applies the electrical signals to the sample in the flow path, detects the electrical signals of the sample, and detects whether there is the single cell in the sample; and a single cell separation control device which outputs a single cell separation control signal when the single cell is detected by the detection electrode.