Automated measurement of diatom size

Sarah A. Spaulding, David H. Jewson, Rebecca J. Bixby, Harry Nelson, Diane M. McKnight

Limnol. Oceanogr. Methods 10:882-890 (2012) | DOI: 10.4319/lom.2012.10.882

ABSTRACT: Size analysis of diatom populations has not been widely considered, but it is a potentially powerful tool for understanding diatom life histories, population dynamics, and phylogenetic relationships. However, measuring cell dimensions on a light microscope is a time-consuming process. An alternative technique has been developed using digital flow cytometry on a FlowCAM® (Fluid Imaging Technologies) to capture hundreds, or even thousands, of images of a chosen taxon from a single sample in a matter of minutes. Up to 30 morphological measures may be quantified through post-processing of the high resolution images. We evaluated FlowCAM size measurements, comparing them against measurements from a light microscope. We found good agreement between measurement of apical cell length in species with elongated, straight valves, including small Achnanthidium minutissimum (11-21 μm) and large Didymosphenia geminata (87-137 μm) forms. However, a taxon with curved cells, Hannaea baicalensis (37-96 μm), showed differences of ~ 4 μm between the two methods. Discrepancies appear to be influenced by the choice of feret or geodesic measurement for asymmetric cells. We describe the operating conditions necessary for analysis of size distributions and present suggestions for optimal instrument conditions for size analysis of diatom samples using the FlowCAM. The increased speed of data acquisition through use of imaging flow cytometers like the FlowCAM is an essential step for advancing studies of diatom populations.