Abstract Background and Aims The line intersect method is widely used in rhizotron and minirhizotron studies to quantify roots and study cultivar and treatment differences in root growth. We investigated ways to optimize the line intersect method and root depth measurements with respect to data variability and the time spent on counting roots. Methods Root intensity was measured with three different grid patterns and different lengths of counting line on 2 m long transparent tube rhizotrons. Rooting depth was recorded by measuring the depth of the deepest root and by measuring the depth below which 5, 10 and 25 roots were observed. Results For root intensity the coefficient of variation (CV) was reduced 10-50 percentage points for grids that distributed counting lines equally across the measured area compared to using a restricted centralized area. In addition, the CV approached an asymptote of around 40 % when more than 50 root intersections per grid were observed. Further we show that recordings of the deepest root gave the most variance and least difference between means with a p-value of 0.65 for difference between cultivars. In contrast, a significant difference between cultivar rooting depths (p = 0.01) was found when using the depth below which 25 roots were observed. Conclusion We propose the use of grid designs adapted to different root densities to decrease time spent on counting roots at high root intensities, and minimize data variability at low root intensities. Further on rooting depth measurements including more roots may be a more useful parameter statistically to reveal variety or treatment differences in rooting depth.