The present chapter has the aim to considerate the most significant aspects of chlorophyll (Chl) applications in the ecological study of fresh waters on the example of the Volga River reservoirs. Throughout the cascade of seven large reservoirs, Chl varied in wide range from 2.5–9 to over 100 μg/L with mean values of 16.5–41.2, 6.7–44.0, and 3.6–10.6 μg/L in the Upper, Middle, and Lower Volga, respectively. Mean Chl values that constantly decrease from the Upper Volga to Lower Volga, characterize Ivankovo, Uglich, and Cheboksary reservoirs as eutrophic, Saratov and Volgograd reservoirs as mesotrophic, while Gorky and Kuibyshev reservoirs in some years are mesotrophic or eutrophic. Chl seasonal dynamics in the Rybinsk reservoir that is dynamics of phytoplankton biomass, is characterized by spring, summer, and, in some years, autumn maxima. Water temperature and water regime of the reservoir are the main factors in Chl dynamics. Years with low-water conditions are favorable for the high Chl concentrations and intensive development of algae. Seasonally average Chl that make from 5 to 22 μg/L during 1969–2019, show variations in trophic state of reservoir from mesotrophic (Chl < 10 μg/L), to moderately eutrophic (10–15 μg/L), and eutrophic (15–22 μg/L).