Waterlogging can reduce crop yield by 20%–50% or more, and lack of efficient selection methods is an obstacle in plant breeding. The methods currently used are mainly indices based on germination ability in Petri dishes and leaf chlorosis in plants grown in waterlogged soils. Cultivation in oxygen-depleted nutrient solution is the ultimate waterlogging system. Therefore methods based on root growth inhibition and on fluorescence in plant material hydroponically grown in oxygen-depleted solution were evaluated against data on biomass accumulation in waterlogged soils. Both traits were correlated with waterlogging tolerance in soil, but since it was easier to measure fluorescence, this method was further evaluated. A selection of F2 plants with high and low fluorescence revealed a small but significant screening effect in F3 plants. A test of 175 Nordic cultivars showed large variations in chlorophyll fluorescence in leaves from oxygen-stressed seedlings, indicating that adaptation to waterlogging has gradually improved over the past 40–50 years with the introduction of new cultivars onto the market. However, precipitation also increased during the period and new cultivars may have inadvertently been adapted to this while breeding barley for grain yield. The results suggest that the hydroponic method can be used for screening barley populations, breeding lines or phenotyping of populations in developing markers for quantitative trait loci.