Imaging Flow Cytometry

Mark Hildebrand, Aubrey Davis, Raffaela Abbriano, Haley R. Pugsley, Jesse C. Traller, Sarah R. Smith, Roshan P. Shrestha, Orna Cook, Eva L. Sánchez-Alvarez, Kalpana Manandhar-Shrestha, Benjamin Alderete, Hildebrand, Mark, Davis, Aubrey, Abbriano, Raffaela, Pugsley, Haley R., Traller, Jesse C., Smith, Sarah R., Shrestha, Roshan P., Cook, Orna, Sánchez-Alvarez, Eva L., Manandhar-Shrestha, Kalpana, Alderete, Benjamin

The ability to image large numbers of cells at high resolution enhances flow cytometric analysis of cells and cell populations. In particular, the ability to image intracellular features adds a unique aspect to analyses, and can enable correlation between molecular phenomena resulting in alterations in cellular phenotype. Unicellular microalgae are amenable to high-throughput analysis to capture the diversity of cell types in natural samples, or diverse cellular responses in clonal populations, especially using imaging cytometry. Using examples from our laboratory, we review applications of imaging cytometry, specifically using an Amnis(®) ImageStream(®)X instrument, to characterize photosynthetic microalgae. Some of these examples highlight advantages of imaging flow cytometry for certain research objectives, but we also include examples that would not necessarily require imaging and could be performed on a conventional cytometer to demonstrate other concepts in cytometric evaluation of microalgae. We demonstrate the value of these approaches for (1) analysis of populations, (2) documentation of cellular features, and (3) analysis of gene expression.