Cell Separation

Part 1 Preparation of single cell suspensions: introduction; methods of dispersal; viability measurements; separation of viable and non-viable cells; cell characterization. Part 2 Fractionation of cells by sedimentation methods: introduction; the theory of sedimentation; separation media; solutions for iso-osmotic gradients; preparation of iso-osmotic gradients; choice of separation method; separation of cells on the basis of size; separation of cells on the basis of density; conclusions. Part 3 Centrifugal elutriation: introduction; principles; equipment; the elutriation procedure; applications; advantages and disadvantages; conclusions; simulation of centrifugal elutriation for the Beckman JE-6B chamber. Part 4 Separation and fractionation of cells by partitioning in aqueous two-phase systems: introduction; theoretical background and its impact on methodology; practical aspects of phase partitioning; selected applications; conclusions; manually opeated countercurrent distribution apparatus. Part 5 Separation of cells by flow cytometry: introduction; the basic instrument; cell sorting by droplet deflection; other flow sorters. Part 6 Immunomethods - magnetic, column and panning techniques: introduction; column and panning methods; reagents for immunoseparation methods; general cell separation using immunomagnetic beads; functional approaches. Part 7 Separation of cells using free flow electrophoresis: introduction; buffers; preparation of cell samples for free flow electrophoresis; preparation and application of free flow electrophoresis for separation of whole cells; evaluation of the electrokinetic properties of cells separated by FFE; overcoming problems; combination of FFE with other techniques.