Computer assistance in multiparameter flow microphotometry of mammalian cells

Abstract

Flow system technology enables the biological and medical experimenter to investigate the distribution spectra of various cellular characteristics separately or in parameter combination on the basis of ultrarapid single cell measurements. A typical rate of recognition is about 1000 to 5000 cells per second and the precision of measurements and their statistical relevance has been previously unobtainable. According to the approach of the multiparameter analysis and high data rate, computer assistance in flow system technology is given primary consideration. In this study three different kinds of software controlled modes in data acquisition are demonstrated: normal acquisition and linear accumulation of single parameters, spectra accumulation of two correlated parameters of each single cell and documentation as linear, two‐ or three‐dimensional distribution pattern, and linear accumulation of two correlated parameters simultaneously with their actual signal‐to‐signal ratio. A first attempt to analyze distribution spectra was the application of the entropy of the structure routinely used in cybernetics. This function seems to be a measure for determining the degree of synchrony in an appropriate pretreated cell population. A special mathematical strategy has been applied to the linear spectra of cellular DNA content, whose advantage is the quantitative extraction of the fractions concerning the various phases of the life cycle cells. The validity of this special curve fitting procedure has been proven on various experimental cell populations.