The fluorescence and scattering properties of Prochlorococcus and Synechococcus at Station ALOHA as measured by flow cytometry (termed the FCM phenotype) vary with depth and over a variety of time scales. The variation in FCM phenotypes may reflect population selection or physiological acclimation to local conditions. Observations before, during, and after a storm with deep water mixing show a short-term homogenization of the FCM phenotypes with depth, followed by a return to the stable pattern over the time span of a few days. These dynamics indicate that, within the upper mixed-layer, the FCM phenotype distribution represents acclimation to ambient light. The populations in the pycnocline (around 100 m and below), remain stable and are invariant with light conditions. In samples where both cyanobacteria coexist, fluorescence properties of Prochlorococcus and Synechococcus are tightly correlated providing further evidence that FCM phenotype variability is caused by a common environmental factor or factors. Measurements of the dynamics of FCM phenotypes provide insights into phytoplankton physiology and adaptation. Alternatively, FCM phenotype census of a water mass may provide information about its origin and illumination history.

Optical properties of eight algae species were measured on a flow cytometer. Forward and perpendicular light scatter measurements provide information on the size and shape of algae cells. The intensity of chlorophyll fluorescence varies greatly among the studied algae species and can be used to distinguish them. Measurements of chlorophyll fluorescence after excitation with different wavelengths provide a fluorescence excitation spectrum for each species over the available wavelength range. These spectra reflect the different photosynthetic pigment contents of the species. Staining algae cells with the DNA stains, Hoechst 33342 and DAPI, provides two additional optical parameters t o distinguish algae populations: blue nuclear fluorescence and yellow granular fluorescence. The combination of these optical measurements enables the distinction of each algae species into a small cluster in a hyperspace of parameters. The automation of phytoplankton analysis on the flow cytometer may lead to the rapid and objective assessment of water quality.Key terms: Algae, flow cytometry, chlorophyll fluorescence, light scatter, fluorescence excitation spectrum, Hoechst 33342, DAPI Studies on the species diversity and abundance of the plankton community can be used as a measure of water quality (6, 8). However, the use of this approach has been limited by the amount of time and expertise needed to identify and count hundreds of species in thousands of water samples by light microscopy. Flow cytometry has the advantage over light microscopy in that, with flow cytometry, large numbers of cells, and thus large numbers of samples, can be analyzed in a very short time. In addition, objective, quantifiable criteria for the discrimination of cell populations can be established.Flow cytometry is regularly used in studies of mammalian cells. Cells are suspended in a liquid stream and pass through a laser beam. Light signals containing information over cell size, structure and chemical composition are emitted from each cell and can be measured. Using measurements of the binding of specific fluorescent dyes and the light scattered by cells, biologists have been able to measure a variety of biological properties and to differentiate among functional populations of cells. These same principles and techniques can be applied to the analysis of plankton samples.

The large-scale structure of chromatin corresponding to G- and R-bands in human G0/G1 interphase nuclei was compared. Fluorescence in situ hybridization (FISH) was used to measure the interphase distance between 42 pairs of probes separated by 0.1-1.5 Mbp. The probe pairs were derived from 21q22.2 and Xp21.3, G-band positive regions, and from 4p16.3, 6p21.3, and Xq28, R-band positive regions. Distributions of measured interphase distances in all regions approximated a Rayleigh distribution, suggesting that the chromatin follows a random-walk path over this range. A linear correlation of mean-square interphase distance and genomic separation, also indicative of random-walk folding, was observed in all regions. The slope of the correlation observed using probes from G-band regions was systematically lower than that from R-band regions. The difference in the slope between Xp21.3 and Xq28 was particularly striking and was observed in normal fibroblast cells, fixed alternatively with methanol and acetic acid or paraformaldehyde, and HeLa cells. These results demonstrate regional differences in large-scale chromosome structure during interphase, with the more openly configured chromatin corresponding to R-bands.

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