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Observation of a brush on the cell surface with the atomic force microscopy (AFM) in vitro is reported. The number of methods to study brushes that coat living cells is limited despite their biological importance. Moreover, it is important to take into account the brush layer when studying cell mechanics. Here the authors present an AFM method to detect the length and grafting density of the brush on viable cells with resolution that considerably surpasses any existing method. The authors demonstrate this method using cultured human cervical epithelial cells, but it can be applied to any type of cell.