AbstractA split‐flow thin (SPLITT) cell with a perpendicular driving force of one gravity has been utilized for the rapid separation of micron‐sized particles from macromolecules. The procedure involves the simultaneous use of two transport mechanisms and thus two operating modes: a sedimentation process controls the displacement of the particles across the thickness of the thin channel, while diffusion controls the displacement of macromolecules. The theoretical equations for these two operating modes are summarized and it is shown how the two modes can be combined to yield specified recovery factors. The theory was tested on a mixture of 10 μm polystyrene latex beads and three different proteins. The observed separation was in excellent agreement with theory. Attempts to fractionate red blood cells and plasma proteins from whole blood were only partially successful as a consequence of the weak sedimentation of red blood cells. Various remedies to this problem are suggested, the most promising of which is the use of a SPLITT cell subject to mild centrifugal fo