Quantification of yeast flocculation under defined conditions will help to understand the physical mechanisms of the flocculation process used in beer fermentation. Flocculation was quantified by measuring the size of yeast flocs and the number of single cells. For this purpose, a method to measure flee size and number of single cells in situ was developed. In this way, it was possible to quantify the actual flocculation during fermentation, without influencing flocculation. The effects of three physical parameters, flee strength, fluid shear, and yeast cell concentration, on flocculation during beer fermentation, were examined. Increasing flee strength results in larger flocs and lower numbers of single cells. If the fluid shear is increased, the size of the flocs decreases, and the number of single cells remains constant at approximately 10% of the total cells present. The cell concentration also influences flocculation, a reduction of 50% in cell concentration leads to a decrease of about 25% in flee size. The number of single cells decreases in linear proportion to the cell concentration. This means that, during yeast settling at full scale, the number of single cells decreases. The results of this study are used in a model for yeast flocculation. With respect to full scale fermentation the effect of cell concentration will play an important role, for flocculation and sedimentation will occur simultaneously leading to a quasi steady state between these phenomena.