title: Three-dimensional imaging flow cytometry through light-sheet fluorescence microscopy.
abstract: Flow cytometry is the tool of choice for high-speed acquisition and analysis of large cell populations, with the tradeoff of lacking intracellular spatial information. Although in the last decades flow cytometry systems that can actually acquire two-dimensional spatial information were developed, some of the limitations remained though, namely constrains related to sample size and lack of depth or dynamic information. The combination of fluidics and light-sheet illumination has the potential to address these limitations. By having cells travelling with the flowing sheath one can, in a controlled fashion, force them at constant speed through the light-sheet enabling the synchronized acquisition of several optical sections, that is, three-dimensional imaging. This approach has already been used for imaging cellular spheroids, plankton, and zebra-fish embryos. In this review, we discuss the known solutions and standing challenges of performing three-dimensional high-throughput imaging of multicellular biological models using fluidics, while retaining cell and organelle-level resolution. © 2017 International Society for Advancement of Cytometry.
authors.affiliation: Imaging and Cytometry Unit (UIC), Instituto Gulbenkian de Ciência, Oeiras, 2780-156, Portugal.|ICFO-Institut de Ciencies Fotóniques, BIST-Barcelona Institute of Science and Technology, 08860 Castelldefels, Barcelona, Spain.
authors.forename: Emilio J
authors.initials: EJ
authors.lastname: Gualda
authors.affiliation: Imaging and Cytometry Unit (UIC), Instituto Gulbenkian de Ciência, Oeiras, 2780-156, Portugal.
authors.forename: Hugo
authors.initials: H
authors.lastname: Pereira
authors.affiliation: Imaging and Cytometry Unit (UIC), Instituto Gulbenkian de Ciência, Oeiras, 2780-156, Portugal.
authors.forename: Gabriel G
authors.initials: GG
authors.lastname: Martins
authors.affiliation: Imaging and Cytometry Unit (UIC), Instituto Gulbenkian de Ciência, Oeiras, 2780-156, Portugal.
authors.forename: Rui
authors.initials: R
authors.lastname: Gardner
authors.affiliation: Imaging and Cytometry Unit (UIC), Instituto Gulbenkian de Ciência, Oeiras, 2780-156, Portugal.
authors.forename: Nuno
authors.initials: N
authors.lastname: Moreno
mesh_terms: D000818:Animals; D005434:Flow Cytometry / Q000379:methods*; D057166:High-Throughput Screening Assays; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010933:Plankton / Q000648:ultrastructure; D018874:Spheroids, Cellular / Q000648:ultrastructure; D015027:Zebrafish
keywords: high throughput microscopy; imaging flow cytometry; light sheet fluorescence microscopy