Colloidal organic matter (COM) exists ubiquitously in natural waters, and plays a pivotal role in biogeochemical cycles. Lake Taihu is confronted with severe eutrophication and algal blooms. Field investigations were carried out in Lake Taihu and two inflow rivers for two years to identify the source of COM and explore the bioavailability of COM to algae. Compositions and sources of COM were analyzed through UV absorption coefficients and parallel factor analysis of three-dimensional excitation emission matrix spectra (3D EEMs-PARAFAC). Absorption coefficient a(350) of COM in rivers surpassed that in the lake, SUVA254 exhibited noteworthy seasonal variations, and SR indicated a stable autochthonous characteristic. Three components were identified by PARAFAC analysis: tryptophan-like (C1), humic-like (C2), and tyrosine-like (C3). Fluorescence indexes showed that C1 and C3 were mainly autochthonously produced and C2 was derived from allochthonous inputs. The bioassays were conducted to assess the bioavailability of COM to the dominant species, Microcystis aeruginosa. The results showed that colloids supplied rich carbon sources and trace metals, supporting the growth of Microcystis aeruginosa. The specific growth rate, Chl a content, and biological increment of Microcystis aeruginosa in 50 % colloid-added treatment exceeded those in 20 % and 75 % treatments. Simultaneously, the effective quantum yield (Fv/Fm) and the apparent photosynthetic electron transport rate (ETRmax) demonstrated that the photosynthetic activity of Microcystis aeruginosa was higher in 20 % treatment. These findings help in understanding the intricate mechanisms of COM in lake ecosystems, and provide the robust scientific foundation for water management and protection in Lake Taihu.