Simultaneous dual-color light sheet fluorescence imaging flow cytometry for highthroughput marine phytoplankton analysis

This paper reports the development of a dual-color light sheet fluorescence imaging flow cytometer exclusively designed for rapid phytoplankton analysis. By simultaneously exciting chlorophyll and phycoerythrin fluorescence, the system is enabled to discriminate phycoerythrin-containing and phycoerythrin-lacking phytoplankton groups through simultaneous two-channel spectral imaging-in-flow. It is demonstrated the system has good sensitivity and resolution to detect picophytoplankton down to the size of ∼1μm, high throughput of 1.3 × 10⁵cells/s and 5 × 10³cells/s at 100μL/min and 3mL/min volume flow rates for cultured picophytoplankton and nanophytoplankton detection, respectively, and a broad imaging range from ∼1μm up to 300μm covering most marine phytoplankton cell sizes with just one 40 × objective. The simultaneous realization of high resolution, high sensitivity and high throughput with spectral resolving power of the system is expected to promote the technology towards more practical applications that demand automated phytoplankton analysis.