Cameraless high-throughput three-dimensional imaging flow cytometry

Today’s imaging flow cytometer (IFC) systems are limited by the projection problem: collapsing three-dimensional (3D) information onto a two-dimensional (2D) image causes a lack of tomographic 3D resolution and reduced information content, limiting the reliability of spot counting or co-localization crucial to cell phenotyping. We present 3D imaging flow cytometry as a solution to the problem. Our high-throughput 3D cell imager based on optical sectioning microscopy combines orthogonal light-sheet scanning illumination with our previous spatiotemporal transformation detection to produce 3D cell image reconstruction from a cameraless single-pixel photodetector readout. We demonstrate this capability by cocapturing 3D fluorescence and label-free side-scattering images of single cells in flow with a throughput of approximately 500 cells per second.

[1]  Jindong Tian,et al.  Quantitative refractive index distribution of single cell by combining phase-shifting interferometry and AFM imaging , 2017, Scientific Reports.

[2]  A. Nakamura,et al.  Use of the γ-H2AX assay to monitor DNA damage and repair in translational cancer research. , 2012, Cancer letters.

[3]  Yu-Hwa Lo,et al.  Imaging Cells in Flow Cytometer Using Spatial-Temporal Transformation , 2015, Scientific Reports.

[4]  Pasquale Memmolo,et al.  Tomographic flow cytometry by digital holography , 2016, Light: Science & Applications.

[5]  R. Medema,et al.  Chromosome Segregation Errors as a Cause of DNA Damage and Structural Chromosome Aberrations , 2011, Science.

[6]  Stephan Preibisch,et al.  OpenSPIM: an open-access light-sheet microscopy platform , 2013, Nature Methods.

[7]  John T. Chang,et al.  Asymmetric T Lymphocyte Division in the Initiation of Adaptive Immune Responses , 2007, Science.

[8]  Fumihito Arai,et al.  Intelligent Image-Activated Cell Sorting , 2018, Cell.

[9]  Lucas Pelkmans,et al.  Using Cell-to-Cell Variability—A New Era in Molecular Biology , 2012, Science.

[10]  O. Laerum,et al.  Clinical application of flow cytometry: a review. , 1981, Cytometry.

[11]  Seng H. Cheng,et al.  Intracellular protein trafficking defects in human disease. , 1992, Trends in cell biology.

[12]  P. H. Yap,et al.  Cell refractive index for cell biology and disease diagnosis: past, present and future. , 2016, Lab on a chip.

[13]  Joseph M. Martel,et al.  Three-Dimensional Holographic Refractive-Index Measurement of Continuously Flowing Cells in a Microfluidic Channel. , 2014, Physical review applied.

[14]  Elina Ikonen,et al.  When intracellular logistics fails - genetic defects in membrane trafficking , 2006, Journal of Cell Science.

[15]  Natalia Gomez-Navarro,et al.  Protein sorting at the ER–Golgi interface , 2016, The Journal of cell biology.

[16]  A. Shibata,et al.  3D-structured illumination microscopy reveals clustered DNA double-strand break formation in widespread γH2AX foci after high LET heavy-ion particle radiation , 2017, Oncotarget.

[17]  Christophe E. Redon,et al.  H2AX: functional roles and potential applications , 2009, Chromosoma.

[18]  L. Kaestner,et al.  3D tomography of cells in micro-channels , 2017, 1709.05686.

[19]  A. Ben-Yakar,et al.  Line excitation array detection fluorescence microscopy at 0.8 million frames per second , 2018, Nature Communications.

[20]  Bahram Jalali,et al.  High-throughput single-microparticle imaging flow analyzer , 2012, Proceedings of the National Academy of Sciences.

[21]  Antoni Ribas,et al.  Single-cell analysis tools for drug discovery and development , 2015, Nature Reviews Drug Discovery.

[22]  I. Vorobjev,et al.  Imaging Flow Cytometry , 2012, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[23]  Hui Ling Chen,et al.  CTCF Mediates Interchromosomal Colocalization Between Igf2/H19 and Wsb1/Nf1 , 2006, Science.

[24]  B. Snijder,et al.  Origins of regulated cell-to-cell variability , 2011, Nature Reviews Molecular Cell Biology.

[25]  Lani F. Wu,et al.  Cellular Heterogeneity: Do Differences Make a Difference? , 2010, Cell.

[26]  J. Lichtman,et al.  Optical sectioning microscopy , 2005, Nature Methods.

[27]  William G Telford,et al.  Flow cytometry of fluorescent proteins. , 2012, Methods.

[28]  Kornelia Polyak,et al.  Cellular heterogeneity and molecular evolution in cancer. , 2013, Annual review of pathology.

[29]  Christophe Zimmer,et al.  FISH-quant: automatic counting of transcripts in 3D FISH images , 2013, Nature Methods.

[30]  Jianglai Wu,et al.  A light sheet based high throughput 3D-imaging flow cytometer for phytoplankton analysis. , 2013, Optics express.

[31]  Jordan R. Myers,et al.  Ultra-High Resolution 3D Imaging of Whole Cells , 2016, Cell.

[32]  William E. Ortyn,et al.  Cellular image analysis and imaging by flow cytometry. , 2007, Clinics in laboratory medicine.

[33]  N. Dovichi,et al.  Simplified sheath flow cuvette design for ultrasensitive laser induced fluorescence detection in capillary electrophoresis. , 2012, The Analyst.

[34]  Y. Lo,et al.  Review: imaging technologies for flow cytometry. , 2016, Lab on a chip.

[35]  R. Horisaki,et al.  Ghost cytometry , 2018, Science.