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12 October 1995
Below is a description of the analysis (modified from DuRand, M.D.
and R.J. Olson, in press, Contributions of phytoplankton light
scattering and cell concentration changes to diel variations in beam
attenuation in the equatorial Pacific from flow cytometric measurements
of pico-, ultra-, and nanoplankton, Deep-Sea Research):
The eukaryotic phytoplankton were analyzed immediately on board the
ship using an EPICS V flow cytometer modified to analyze 50 ml seawater
samples at 5-10 ml min-1 (Olson et al., 1991, 1993). Samples were kept
at room temperature in the dark until analysis; the surface samples were
analyzed first (the upper 45 m samples were analyzed within an hour)
and all depths were completed within two hours of collection. During
each cruise the phytoplankton groups were sorted on the flow cytometer
and examined microscopically. Typically, the most abundant population
of eukaryotic phytoplankton consisted of 1-2 um-diameter coccoid cells
(referred to here as the ultraphytoplankton). The population referred
to here as the nanophytoplankton primarily consisted of 2-3 um diameter
coccoid cells, but also included cells as large as 20 um.
The picoplankton Synechococcus and Prochlorococcus were analyzed
on shore from samples preserved with 0.125% glutaraldehyde and
stored in liquid nitrogen (Vaulot et al., 1989, Olson et al., 1993),
using a high sensitivity configuration of an EPICS 753 flow cytometer
(Olson et al., 1993). Synechococcus were distinguished by their high
phycoerythrin fluorescence and Prochlorococcus by their low forward
light scatter and chlorophyll fluorescence.
Olson, R.J., E.R. Zettler, S.W. Chisholm and J.A. Dusenberry. (1991) Advances
in oceanography through flow cytometry, pp. 351-399 in S. Demers (ed.),
Particle Analysis in Oceanography, NATO ASI Series G 27, Springer-Verlag,
Berlin.
Olson, R.J., E.R. Zettler and M.D. DuRand. (1993) Phytoplankton analysis
using flow cytometry, pp. 175-186 in P.F. Kemp, B.F. Sherr, E.B. Sherr
and J.J. Cole (eds.), Handbook of Methods in Aquatic Microbial Ecology,
Lewis Publishers, Boca Raton.
Vaulot, D., C. Courties and F. Partensky. (1989) A simple method to preserve
oceanic phytoplankton for flow cytometric analyses. Cytometry, 10,
629-635.