Three‐dimensional imaging flow cytometry through light‐sheet fluorescence microscopy

• Published in: Cytometry. Part A Vol. 91; no. 2; pp. 144 - 151
• Main Authors: Gualda, Emilio J., Pereira, Hugo, Martins, Gabriel G., Gardner, Rui, Moreno, Nuno
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.02.2017
• Subjects: Acquisitions & mergers; Animals; Biological models (mathematics); Cytometry; Embryos; Flow cytometry; Flow Cytometry - methods; Fluidics; Fluorescence; Fluorescence in situ hybridization; Fluorescence microscopy; High speed; high throughput microscopy; High-Throughput Screening Assays; Illumination; Imaging; imaging flow cytometry; Imaging, Three-Dimensional - methods; light sheet fluorescence microscopy; Microscopy; Microscopy, Fluorescence - methods; Plankton; Plankton - ultrastructure; Populations; Spheroids; Spheroids, Cellular - ultrastructure; Three dimensional flow; Three dimensional imaging; Three dimensional models; Zebrafish; imaging flow cytometry; light sheet fluorescence microscopy; high throughput microscopy
• ISSN: 1552-4922; 1552-4930
• DOI: 10.1002/cyto.a.23046

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