|flow cytophotometry -->|
Complex computerised instruments are used to pass a monocellular stream of cells, platelets or other microscopic particulate elements through a beam of laser light. The cells are categorised first by size and then computer analysed to sort the mixture of cellular elements into cell type by size.
Cells are labelled with fluorescent dye and then passed, in suspending medium, through a narrow dropping nozzle so that each cell is in a small droplet. A laser based detector system is used to excite fluorescence and droplets with positively fluorescent cells are given an electric charge. Charged and uncharged droplets are separated as they fall between charged plates and so collect in different tubes. The machine can be used either as an analytical tool, counting the number of labelled cells in a population or to separate the cells for subsequent growth of the selected population. Further sophistication can be built into the system by using a second laser system at right angles to the first to look at a second fluorescent label or to gauge cell size on the basis of light scatter. The great strength of the system is that it looks at large numbers of individual cells and makes possible the separation of populations with, for example: particular surface properties.
Tabulation of counted data in conjunction with size analysis enables determination of relative percentages of each specific cellular subset for which monoclonal antibody conjugates are utilised, even when the size of the cell is identical to other subset species.
(01 Dec 1998)
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