Azotobacter is aerobic bacteria. and Cladophora is green alga. Engelmann used a prism to split light into its spectral components, and then illuminated a green alga, Cladophora, placed in a suspension of aerobic bacteria. The bacteria were used to detect the sites of oxygen evolution. He observed that aerobic bacteria accumulated mainly in the region of blue and red light of the split spectrum thus giving the first action spectrum of photosynthesis.