Microscope-based light gradient generation for quantitative growth studies of photosynthetic micro-organisms

Photosynthetic micro-organisms are equipped with molecular machineries that are designed to transform light into chemical or bioenergy, and help shape and balance the ecosystem of all life forms on earth. Recently, aquatic ecosystems have been disrupted by climate change, which leads to the frequent occurrence of harmful algal blooms (HABs). HABs endanger drinking water resources and harm the fishing and coastal recreation industries. Despite its urgency, mechanistic understanding of how key biophysical and biochemical parameters impact algal growth is largely unexplored. In this article, we developed a microscope-based light gradient generator for studies of photosynthetic micro-organisms under well-defined light intensity gradients. This technology utilized a commercially available microscope, allowed for controlled light exposure and imaging of cells on the same microscope platform, and can be integrated with any micrometer-scale device. Using this technology, we studied the role of light intensity in the growth of photosynthetic micro-organisms. A parallel study was also carried out using a 96-well plate. Our work revealed that the growth rate of the microalgae/cyanobacteria was significantly regulated by the light intensity and followed Monod or van Oorschot kinetic models. The measured half-saturation constants were compared with those obtained in macro-scale devices, and indicated that shading, light spectrum, and temperature may all play important roles in the light sensitivity of photosynthetic micro-organisms. This work highlighted the importance of analytical tools for quantitative understanding of biophysical parameters in the growth of photosynthetic micro-organisms, and knowledge learned will be critical in the design of future technologies for managing algal blooms or optimizing bioenergy production.