Lipid content and lipid class composition were determined in stream periphyton and the filamentous green algae Cladophora sp. and Spirogyra sp. Sterols and phospholipids were compared to chlorophyll a (chi a) as predictors of biomass for stream periphyton and algae. Chlorophyll a, phospholipids, and sterols were each highly correlated with ash-free dry mass (AFDM) (r(2) > 0.98). Stream periphyton exposed naturally to high light (HL) and low light (LL) had chi a concentrations (mu g chl a. mg(-1) AFDM) of 7.9 +/- 0.7 and 12.4 +/- 2.9, respectively, while the sterol concentrations of these HL and LL stream periphyton (1.6 +/- 0.4) were not significantly different (P > 0.05). Periphyton exposed to an irradiance of 300 mu mol photons.m(-2).s(-1) in the laboratory for 60 h had 5.6 +/- 0.55 mu g chl a.mg(-1) AFDM, but the same periphyton exposed to 2% incident light for the same amount of time had 11.0 +/- 0.56 mu g chl a.mg(-1) AFDM. Sterol concentrations in these periphyton communities remained unchanged (1.5 +/- 0.3 mu g.mg(-1) AFDM). Similar results (i.e. changes in chi a but stability of sterol concentrations in response to irradiance changes) were also found for Cladophora and Spirogyra in laboratory experiments. Sterols can be quantified rapidly from a few milligrams of algae and appear to be a useful predictor of eukaryote biomass, whereas cellular levels of chl a vary substantially with light conditions. Phospholipids (or phospholipid fatty acids) are considered to be a reliable measure of viable microbial biomass. Nevertheless, phospholipid content varied substantially and unpredictably among algae and periphyton under different light regimes. Irradiance also had a significant effect on storage lipids: HL Cladophora and HL periphyton had 2 x and 5 x greater concentrations of triacylglycerols, respectively, compared to their LL forms. HL and LL algae also differed in the concentration of several major fatty acids. These light-induced changes in algal lipids and fatty acids have important implications for grazers.