Environmental health: flow cytometric methods to assess our water world.
Flow cytometry/cell sorting in aquatic sciences has been driven in two directions. The frontier directions are on shipboard and shore-based. On the one hand, the rapid analytical technique has been taken on shipboard to provide a real-time assessment of the particles and phytoplankton in water masses. These data also give information on the amount of vertical mixing and advection, and denote fronts between two or more water masses. There is an optical characterization (based on sizes, numbers, and pigment groups) of the individual primary producers, as well as detritus and suspended sediments. An optical-closure question is being addressed: "Does the total optical signal equal the sum of the parts?" Additionally, associations with chemical and physical oceanographic features are readily accomplished. A "census" of thousands of phytoplankton cells is obtained and can be mapped. Scientists are able to identify "who is where?" Such data are critical to understand the optical-feedback loop or the so-called photon-budget-in-the-sea, which in turn controls the rates at which growth processes occur in nature. On the other hand, an in-depth understanding is sought as to how particle size, shape, refractive index, nutritional status (nutrient and/or light limitation), growth dynamics, and cell cycle combine to control the optics (light scatter and fluorescence at the moment, and ideally absorption as well) or the photon-budget-of-the-cell. For this purpose, a shore-based facility associated with a diverse collection of phytoplankton is ideal. The development at Bigelow Laboratory of the Jane J. MacIsaac Facility is to provide services for the oceanographic community. Association and co-location with the Provasoli-Guillard Center for Culture of Marine Phytoplankton is key. Visitors are trained and given access to state-of-the-art instrumentation. Visiting investigators have available "the tropical, temperate, and polar seas" in concentrated form, as marine phytoplankton isolated worldwide and maintained as living clonal cultures. In this way, frontline cell biology questions can be addressed. The relentless exploration of standards and controls appropriate for the aquatic community must be continued. An intercalibration effort is a vital step. It is only with the widespread acceptance of particular reference materials and uniform optical filters among research groups utilizing FCM that comparable data sets describing aquatic particle distributions will be possible. For a global science, this strategy is imperative.
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J.J. MacIsaac Flow Cytometry/Sorting Facility, Bigelow Laboratory for Ocean Sciences, West Boothbay Harbor, Maine 04575.
D003124:Colorimetry / Q000379:methods; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D008386:Marine Biology / Q000295:instrumentation / Q000379:methods*; D009391:Nephelometry and Turbidimetry / Q000379:methods; D010788:Photosynthesis; D010839:Phytoplankton / Q000145:classification / Q000502:physiology / Q000528:radiation effects; D010860:Pigments, Biological / Q000032:analysis; D013685:Telecommunications
D005456:Fluorescent Dyes; D010860:Pigments, Biological