In recent years, in-situ fluorometers have been extensively deployed to monitor cyanobacteria in near real-time. Acceptable accuracy can be achieved between measured pigments and cyanobacteria biovolume provided the cyanobacteria species are known. However, cellular photosynthetic pigment content and measurement interferences are site and species specific and can dramatically affect sensor reliability. We quantified the accuracy of an in-situ fluorometer compared with traditional methods using mono- and mixed cultures of four different cyanobacterial species. We found: (1) lower pigment content in cultures in stationary phase, (2) higher precision with the sensor compared to traditional pigment quantification methods of measuring phycocyanin and chlorophyll a, (3) species-specific relationships between sensor readings and measurements related to biovolume, (4) overestimation of pigments in mixed compared with mono cultures, (5) dissolved organic matter causing a loss in signal proportional to its degree of aromaticity, and (6) potential to quantify the degree of cell lysis with a fluorescent dissolved organic matter sensor. This study has provided important new information on the strengths and limitations of fluorescence sensors. The sensor readings can provide accurate biovolume quantification and species determination for a number of bloom-forming species when sensors are properly compensated and calibrated.