On-chip light-sheet fluorescence imaging flow cytometry at a high flow speed of 1 m/s.

We present on-chip fluorescence imaging flow cytometry by light-sheet excitation on a mirror-embedded microfluidic chip. The method allows us to obtain microscopy-grade fluorescence images of cells flowing at a high speed of 1 m/s, which is comparable to the flow speed of conventional non-imaging flow cytometers. To implement the light-sheet excitation of flowing cells in a microchannel, we designed and fabricated a mirror-embedded PDMS-based microfluidic chip. To show its broad utility, we used the method to classify large populations of microalgal cells (Euglena gracilis) and human cancer cells (human adenocarcinoma cells). Our method holds promise for large-scale single-cell analysis.

[1]  Chun-Hao Chen,et al.  Microfluidics and photonics for Bio‐System‐on‐a‐Chip: A review of advancements in technology towards a microfluidic flow cytometry chip , 2008, Journal of biophotonics.

[2]  R. Schantz,et al.  Changes in amino acid and peptide composition of Euglena gracilis cells during chloroplast development , 1975 .

[3]  Andrew J. deMello,et al.  High-Throughput Multi-parametric Imaging Flow Cytometry , 2017 .

[4]  G. Whitesides,et al.  Microfluidic devices fabricated in Poly(dimethylsiloxane) for biological studies , 2003, Electrophoresis.

[5]  G. van den Engh,et al.  Photo-bleaching and photon saturation in flow cytometry. , 1992, Cytometry.

[6]  Cheng Lei,et al.  High-Throughput Accurate Single-Cell Screening of Euglena gracilis with Fluorescence-Assisted Optofluidic Time-Stretch Microscopy , 2016, PloS one.

[7]  T. Laurell,et al.  Two-hundredfold volume concentration of dilute cell and particle suspensions using chip integrated multistage acoustophoresis. , 2012, Lab on a chip.

[8]  Eric P. Y. Chiou,et al.  3D pulsed laser triggered high speed microfluidic fluorescence activated cell sorter , 2011, CLEO: 2013.

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

[10]  P. Mondal,et al.  Light sheet based imaging flow cytometry on a microfluidic platform , 2013, Microscopy research and technique.

[11]  K. Jensen,et al.  Cells on chips , 2006, Nature.

[12]  J. Huisken,et al.  A guide to light-sheet fluorescence microscopy for multiscale imaging , 2017, Nature Methods.

[13]  M. Davidson,et al.  Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination , 2011, Nature Methods.

[14]  Norimichi Tsumura,et al.  Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy , 2016, Nature Microbiology.

[15]  Cheng Lei,et al.  High‐throughput, label‐free, single‐cell, microalgal lipid screening by machine‐learning‐equipped optofluidic time‐stretch quantitative phase microscopy , 2017, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[16]  K. Dholakia,et al.  Light-sheet microscopy using an Airy beam , 2014, Nature Methods.

[17]  Keisuke Goda,et al.  Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting , 2016, Scientific Reports.

[18]  Vincent Studer,et al.  3D high- and super-resolution imaging using single-objective SPIM , 2015, Nature Methods.

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

[20]  Partha Pratim Mondal,et al.  High resolution light-sheet based high-throughput imaging cytometry system enables visualization of intra-cellular organelles , 2014 .