Discrimination of eukaryotic phytoplankton cell types from light scatter and autofluorescence properties measured by flow cytometry

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Abstract
Flow cytometric methods of recognizing several groups of eukaryotic marine phytoplankton were tested using 26 laboratory cultures. Each culture was divided into three aliquots, and these samples were analyzed for 1) Coulter volume; 2) light scatter (magnitude and polarization properties of forward scattered light and magnitude of right-angle scatter light) and autofluorescence emission (phycoerythrin and chorophyll); and 3) autofluorescence excitation (by 488 nm and 515 nm light). Three kinds of cells could be easily distinguished from others in the culture collection: 1) The two cryptophytes and the rhodophyte had high phycoerythrin/chlorophyll ratios; 2) the two coccolithophores depolarized forward scattered lights; and 3) the two pennate diatoms scattered only a relatively small amount of light in the foward direction compared with that at right angles. Mean chlorophyll fluorescence excited by blue light relative to that excited by green light was highest in the four chlorophytes, but there was overlap between some of these and some other kinds of cells. Unresolved cell types included centric diatoms, dinoflagellates, and naked cocclithophores. Forward light scatter and Coulter volume were closely related (except for the pennate diatoms) over a range of about 0.01 to 30 pL (equivalent spherical diameter about 3 to 40 .mu.m), according to a logarithmic function.