Virtual-freezing fluorescence imaging flow cytometry
Open Access
- 5 March 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Communications
- Vol. 11 (1), 1-11
- https://doi.org/10.1038/s41467-020-14929-2
Abstract
By virtue of the combined merits of flow cytometry and fluorescence microscopy, imaging flow cytometry (IFC) has become an established tool for cell analysis in diverse biomedical fields such as cancer biology, microbiology, immunology, hematology, and stem cell biology. However, the performance and utility of IFC are severely limited by the fundamental trade-off between throughput, sensitivity, and spatial resolution. Here we present an optomechanical imaging method that overcomes the trade-off by virtually freezing the motion of flowing cells on the image sensor to effectively achieve 1000 times longer exposure time for microscopy-grade fluorescence image acquisition. Consequently, it enables high-throughput IFC of single cells at >10,000 cells s(-1) without sacrificing sensitivity and spatial resolution. The availability of numerous information-rich fluorescence cell images allows high-dimensional statistical analysis and accurate classification with deep learning, as evidenced by our demonstration of unique applications in hematology and microbiology. High throughput imaging flow cytometry suffers from trade-offs between throughput, sensitivity and spatial resolution. Here the authors introduce a method to virtually freeze cells in the image acquisition window to enable 1000 times longer signal integration time and improve signal-to-noise ratio.Funding Information
- MEXT | Japan Society for the Promotion of Science (Core-to-Core Program, KAKENHI 19H05633, Core-to-Core Program, Core-to-Core Program, Core-to-Core Program, KAKENHI 19H05633)
- Council for Science, Technology and Innovation (ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program, ImPACT Program)
- White Rock Foundation and Precise Measurement Technology Promotion Foundation are a Japanese foundations that provide research grants.
This publication has 39 references indexed in Scilit:
- Inhibition of Dihydroorotate Dehydrogenase Overcomes Differentiation Blockade in Acute Myeloid LeukemiaCell, 2016
- HER2 expression identifies dynamic functional states within circulating breast cancer cellsNature, 2016
- Imaging Flow CytometryPublished by Springer Science and Business Media LLC ,2016
- Imaging flow cytometryJournal of Immunological Methods, 2015
- An MTCH2 pathway repressing mitochondria metabolism regulates haematopoietic stem cell fateNature Communications, 2015
- Real-time deformability cytometry: on-the-fly cell mechanical phenotypingNature Methods, 2015
- Trogocytosis by Entamoeba histolytica contributes to cell killing and tissue invasionNature, 2014
- Quantitative Diagnosis of Malignant Pleural Effusions by Single-Cell MechanophenotypingScience Translational Medicine, 2013
- Cellular Heterogeneity: Do Differences Make a Difference?Cell, 2010
- Identification of a dendritic cell receptor that couples sensing of necrosis to immunityNature, 2009