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Research on microphytobenthos (MPB) photosynthesis has increasingly relied on pulse amplitude modulation fluorometry, often through the generation of light response curves of the relative electron transport of PSII (rETR), generated through the sequential exposure to a range of actinic irradiances. However, fast, vertical phototactic responses by the motile diatoms that dominate these biofilms occur during the generation of sequential light curves. Lightinduced vertical migration is known to confound the characterization of the inherent physiological response of the cells forming the biofilm. The alternative approach of shortening the duration of the light curve protocol prevents the reaching of a physiological steady state and results in light curves heavily dependent on initial light conditions. This study tested the application to MPB of a recently introduced method allowing for the fast generation of nonsequential …