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Three‐dimensional imaging flow cytometry through light‐sheet fluorescence microscopy
Author(s) -
Gualda Emilio J.,
Pereira Hugo,
Martins Gabriel G.,
Gardner Rui,
Moreno Nuno
Publication year - 2017
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.23046
Subject(s) - flow cytometry , cytometry , microfluidics , microscopy , light sheet fluorescence microscopy , multicellular organism , fluidics , flow (mathematics) , fluorescence lifetime imaging microscopy , fluorescence microscope , computer science , biomedical engineering , nanotechnology , biological system , biology , fluorescence , optics , materials science , cell , physics , medicine , genetics , engineering , aerospace engineering , mechanics
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