Abstract

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.

© 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

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2017 (5)

B. Guo, C. Lei, H. Kobayashi, T. Ito, Y. Yalikun, Y. Jiang, Y. Tanaka, Y. Ozeki, and K. Goda, “High-throughput, label-free, single-cell, microalgal lipid screening by machine-learning-equipped optofluidic time-stretch quantitative phase microscopy,” Cytometry A 91(5), 494–502 (2017).
[Crossref] [PubMed]

A. S. Rane, J. Rutkauskaite, A. deMello, and S. Stavrakis, “High-throughput multi-parametric imaging flow cytometry,” Chem 3(4), 588–602 (2017).
[Crossref]

H. Jiang, T. Zhu, H. Zhang, J. Nie, Z. Guan, C.-M. Ho, S. Liu, and P. Fei, “Droplet-based light-sheet fluorescence microscopy for high-throughput sample preparation, 3-D imaging and quantitative analysis on a chip,” Lab Chip 17(13), 2193–2197 (2017).
[Crossref] [PubMed]

R. M. Power and J. Huisken, “A guide to light-sheet fluorescence microscopy for multiscale imaging,” Nat. Methods 14(4), 360–373 (2017).
[Crossref] [PubMed]

E. Zagato, T. Brans, S. Verstuyft, D. van Thourhout, J. Missinne, G. van Steenberge, J. Demeester, S. De Smedt, K. Remaut, K. Neyts, and K. Braeckmans, “Microfabricated devices for single objective single plane illumination microscopy (SoSPIM),” Opt. Express 25(3), 1732–1745 (2017).
[Crossref] [PubMed]

2016 (6)

M. B. M. Meddens, S. Liu, P. S. Finnegan, T. L. Edwards, C. D. James, and K. A. Lidke, “Single objective light-sheet microscopy for high-speed whole-cell 3D super-resolution,” Biomed. Opt. Express 7(6), 2219–2236 (2016).
[Crossref] [PubMed]

C. Lei, T. Ito, M. Ugawa, T. Nozawa, O. Iwata, M. Maki, G. Okada, H. Kobayashi, X. Sun, P. Tiamsak, N. Tsumura, K. Suzuki, D. Di Carlo, Y. Ozeki, and K. Goda, “High-throughput label-free image cytometry and image-based classification of live Euglena gracilis,” Biomed. Opt. Express 7(7), 2703–2708 (2016).
[Crossref] [PubMed]

B. Guo, C. Lei, T. Ito, Y. Jiang, Y. Ozeki, and K. Goda, “High-throughput accurate single-cell screening of Euglena gracilis with fluorescence-assisted optofluidic time-stretch microscopy,” PLoS One 11(11), e0166214 (2016).
[Crossref] [PubMed]

P. Paiè, F. Bragheri, A. Bassi, and R. Osellame, “Selective plane illumination microscopy on a chip,” Lab Chip 16(9), 1556–1560 (2016).
[Crossref] [PubMed]

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

K. Yamada, H. Suzuki, T. Takeuchi, Y. Kazama, S. Mitra, T. Abe, K. Goda, K. Suzuki, and O. Iwata, “Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting,” Sci. Rep. 6(1), 26327 (2016).
[Crossref] [PubMed]

2015 (1)

R. Galland, G. Grenci, A. Aravind, V. Viasnoff, V. Studer, and J.-B. Sibarita, “3D high- and super-resolution imaging using single-objective SPIM,” Nat. Methods 12(7), 641–644 (2015).
[Crossref] [PubMed]

2014 (2)

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light-sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
[Crossref] [PubMed]

R. Regmi, K. Mohan, and P. P. Mondal, “High resolution light-sheet based high-throughput imaging cytometry system enables visualization of intra-cellular organelles,” AIP Adv. 4(9), 097125 (2014).
[Crossref]

2013 (4)

R. Regmi, K. Mohan, and P. P. Mondal, “MRT letter: light sheet based imaging flow cytometry on a microfluidic platform,” Microsc. Res. Tech. 76(11), 1101–1107 (2013).
[Crossref] [PubMed]

J. Wu, J. Li, and R. K. Y. Chan, “A light sheet based high throughput 3D-imaging flow cytometer for phytoplankton analysis,” Opt. Express 21(12), 14474–14480 (2013).
[Crossref] [PubMed]

J. Wu and R. K. Y. Chan, “A fast fluorescence imaging flow cytometer for phytoplankton analysis,” Opt. Express 21(20), 23921–23926 (2013).
[Crossref] [PubMed]

Y. Chen, T.-H. Wu, Y.-C. Kung, M. A. Teitell, and P.-Y. Chiou, “3D pulsed laser-triggered high-speed microfluidic fluorescence-activated cell sorter,” Analyst 138(24), 7308–7315 (2013).
[Crossref] [PubMed]

2012 (2)

M. Nordin and T. Laurell, “Two-hundredfold volume concentration of dilute cell and particle suspensions using chip integrated multistage acoustophoresis,” Lab Chip 12(22), 4610–4616 (2012).
[Crossref] [PubMed]

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

2011 (1)

T. A. Planchon, L. Gao, D. E. Milkie, M. W. Davidson, J. A. Galbraith, C. G. Galbraith, and E. Betzig, “Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination,” Nat. Methods 8(5), 417–423 (2011).
[Crossref] [PubMed]

2008 (1)

J. Godin, C.-H. Chen, S. H. Cho, W. Qiao, F. Tsai, and Y.-H. Lo, “Microfluidics and photonics for Bio-System-on-a-Chip: a review of advancements in technology towards a microfluidic flow cytometry chip,” J. Biophotonics 1(5), 355–376 (2008).
[Crossref] [PubMed]

2007 (1)

D. A. Basiji, W. E. Ortyn, L. Liang, V. Venkatachalam, and P. Morrissey, “Cellular image analysis and imaging by flow cytometry,” Clin. Lab. Med. 27(3), 653–670 (2007).
[Crossref] [PubMed]

2006 (1)

J. El-Ali, P. K. Sorger, and K. F. Jensen, “Cells on chips,” Nature 442, 403–411 (2006).
[Crossref] [PubMed]

2003 (1)

S. K. Sia and G. M. Whitesides, “Microfluidic devices fabricated in Poly(dimethylsiloxane) for biological studies,” Electrophoresis 24(21), 3563–3576 (2003).
[Crossref] [PubMed]

1992 (1)

G. van den Engh and C. Farmer, “Photo-bleaching and photon saturation in flow cytometry,” Cytometry 13(7), 669–677 (1992).
[Crossref] [PubMed]

1975 (1)

R. Schantz, M.-L. Schantz, and H. Duranton, “Changes in amino acid and peptide composition of Euglena gracilis cells during chloroplast development,” Plant Sci. Lett. 5(5), 313–324 (1975).
[Crossref]

1952 (1)

M. Cramer and J. Myers, “Growth and photosynthetic characteristics of euglena gracilis,” Arch. Mikrobiol. 17(1-4), 384–402 (1952).
[Crossref]

Abe, T.

K. Yamada, H. Suzuki, T. Takeuchi, Y. Kazama, S. Mitra, T. Abe, K. Goda, K. Suzuki, and O. Iwata, “Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting,” Sci. Rep. 6(1), 26327 (2016).
[Crossref] [PubMed]

Adam, J.

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

Aravind, A.

R. Galland, G. Grenci, A. Aravind, V. Viasnoff, V. Studer, and J.-B. Sibarita, “3D high- and super-resolution imaging using single-objective SPIM,” Nat. Methods 12(7), 641–644 (2015).
[Crossref] [PubMed]

Ayazi, A.

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

Basiji, D. A.

D. A. Basiji, W. E. Ortyn, L. Liang, V. Venkatachalam, and P. Morrissey, “Cellular image analysis and imaging by flow cytometry,” Clin. Lab. Med. 27(3), 653–670 (2007).
[Crossref] [PubMed]

Bassi, A.

P. Paiè, F. Bragheri, A. Bassi, and R. Osellame, “Selective plane illumination microscopy on a chip,” Lab Chip 16(9), 1556–1560 (2016).
[Crossref] [PubMed]

Betzig, E.

T. A. Planchon, L. Gao, D. E. Milkie, M. W. Davidson, J. A. Galbraith, C. G. Galbraith, and E. Betzig, “Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination,” Nat. Methods 8(5), 417–423 (2011).
[Crossref] [PubMed]

Brackbill, N.

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

Braeckmans, K.

Bragheri, F.

P. Paiè, F. Bragheri, A. Bassi, and R. Osellame, “Selective plane illumination microscopy on a chip,” Lab Chip 16(9), 1556–1560 (2016).
[Crossref] [PubMed]

Brans, T.

Chan, R. K. Y.

Chen, C.-H.

J. Godin, C.-H. Chen, S. H. Cho, W. Qiao, F. Tsai, and Y.-H. Lo, “Microfluidics and photonics for Bio-System-on-a-Chip: a review of advancements in technology towards a microfluidic flow cytometry chip,” J. Biophotonics 1(5), 355–376 (2008).
[Crossref] [PubMed]

Chen, Y.

Y. Chen, T.-H. Wu, Y.-C. Kung, M. A. Teitell, and P.-Y. Chiou, “3D pulsed laser-triggered high-speed microfluidic fluorescence-activated cell sorter,” Analyst 138(24), 7308–7315 (2013).
[Crossref] [PubMed]

Chiou, P.-Y.

Y. Chen, T.-H. Wu, Y.-C. Kung, M. A. Teitell, and P.-Y. Chiou, “3D pulsed laser-triggered high-speed microfluidic fluorescence-activated cell sorter,” Analyst 138(24), 7308–7315 (2013).
[Crossref] [PubMed]

Cho, S. H.

J. Godin, C.-H. Chen, S. H. Cho, W. Qiao, F. Tsai, and Y.-H. Lo, “Microfluidics and photonics for Bio-System-on-a-Chip: a review of advancements in technology towards a microfluidic flow cytometry chip,” J. Biophotonics 1(5), 355–376 (2008).
[Crossref] [PubMed]

Cižmár, T.

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light-sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
[Crossref] [PubMed]

Coll-Lladó, C.

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light-sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
[Crossref] [PubMed]

Cramer, M.

M. Cramer and J. Myers, “Growth and photosynthetic characteristics of euglena gracilis,” Arch. Mikrobiol. 17(1-4), 384–402 (1952).
[Crossref]

Dalgarno, H. I. C.

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light-sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
[Crossref] [PubMed]

Davidson, M. W.

T. A. Planchon, L. Gao, D. E. Milkie, M. W. Davidson, J. A. Galbraith, C. G. Galbraith, and E. Betzig, “Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination,” Nat. Methods 8(5), 417–423 (2011).
[Crossref] [PubMed]

De Smedt, S.

Demeester, J.

deMello, A.

A. S. Rane, J. Rutkauskaite, A. deMello, and S. Stavrakis, “High-throughput multi-parametric imaging flow cytometry,” Chem 3(4), 588–602 (2017).
[Crossref]

Dholakia, K.

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light-sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
[Crossref] [PubMed]

Di Carlo, D.

C. Lei, T. Ito, M. Ugawa, T. Nozawa, O. Iwata, M. Maki, G. Okada, H. Kobayashi, X. Sun, P. Tiamsak, N. Tsumura, K. Suzuki, D. Di Carlo, Y. Ozeki, and K. Goda, “High-throughput label-free image cytometry and image-based classification of live Euglena gracilis,” Biomed. Opt. Express 7(7), 2703–2708 (2016).
[Crossref] [PubMed]

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

Domon, R.

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

Duranton, H.

R. Schantz, M.-L. Schantz, and H. Duranton, “Changes in amino acid and peptide composition of Euglena gracilis cells during chloroplast development,” Plant Sci. Lett. 5(5), 313–324 (1975).
[Crossref]

Edwards, T. L.

El-Ali, J.

J. El-Ali, P. K. Sorger, and K. F. Jensen, “Cells on chips,” Nature 442, 403–411 (2006).
[Crossref] [PubMed]

Fard, A. M.

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

Farmer, C.

G. van den Engh and C. Farmer, “Photo-bleaching and photon saturation in flow cytometry,” Cytometry 13(7), 669–677 (1992).
[Crossref] [PubMed]

Fei, P.

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R. Galland, G. Grenci, A. Aravind, V. Viasnoff, V. Studer, and J.-B. Sibarita, “3D high- and super-resolution imaging using single-objective SPIM,” Nat. Methods 12(7), 641–644 (2015).
[Crossref] [PubMed]

Sollier, E.

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

Sorger, P. K.

J. El-Ali, P. K. Sorger, and K. F. Jensen, “Cells on chips,” Nature 442, 403–411 (2006).
[Crossref] [PubMed]

Stavrakis, S.

A. S. Rane, J. Rutkauskaite, A. deMello, and S. Stavrakis, “High-throughput multi-parametric imaging flow cytometry,” Chem 3(4), 588–602 (2017).
[Crossref]

Studer, V.

R. Galland, G. Grenci, A. Aravind, V. Viasnoff, V. Studer, and J.-B. Sibarita, “3D high- and super-resolution imaging using single-objective SPIM,” Nat. Methods 12(7), 641–644 (2015).
[Crossref] [PubMed]

Sugawara, M.

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

Sun, X.

Suzuki, H.

K. Yamada, H. Suzuki, T. Takeuchi, Y. Kazama, S. Mitra, T. Abe, K. Goda, K. Suzuki, and O. Iwata, “Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting,” Sci. Rep. 6(1), 26327 (2016).
[Crossref] [PubMed]

Suzuki, K.

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

K. Yamada, H. Suzuki, T. Takeuchi, Y. Kazama, S. Mitra, T. Abe, K. Goda, K. Suzuki, and O. Iwata, “Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting,” Sci. Rep. 6(1), 26327 (2016).
[Crossref] [PubMed]

C. Lei, T. Ito, M. Ugawa, T. Nozawa, O. Iwata, M. Maki, G. Okada, H. Kobayashi, X. Sun, P. Tiamsak, N. Tsumura, K. Suzuki, D. Di Carlo, Y. Ozeki, and K. Goda, “High-throughput label-free image cytometry and image-based classification of live Euglena gracilis,” Biomed. Opt. Express 7(7), 2703–2708 (2016).
[Crossref] [PubMed]

Suzuki, Y.

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

Takeuchi, T.

K. Yamada, H. Suzuki, T. Takeuchi, Y. Kazama, S. Mitra, T. Abe, K. Goda, K. Suzuki, and O. Iwata, “Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting,” Sci. Rep. 6(1), 26327 (2016).
[Crossref] [PubMed]

Tanaka, Y.

B. Guo, C. Lei, H. Kobayashi, T. Ito, Y. Yalikun, Y. Jiang, Y. Tanaka, Y. Ozeki, and K. Goda, “High-throughput, label-free, single-cell, microalgal lipid screening by machine-learning-equipped optofluidic time-stretch quantitative phase microscopy,” Cytometry A 91(5), 494–502 (2017).
[Crossref] [PubMed]

Teitell, M. A.

Y. Chen, T.-H. Wu, Y.-C. Kung, M. A. Teitell, and P.-Y. Chiou, “3D pulsed laser-triggered high-speed microfluidic fluorescence-activated cell sorter,” Analyst 138(24), 7308–7315 (2013).
[Crossref] [PubMed]

Tiamsak, P.

Tsai, F.

J. Godin, C.-H. Chen, S. H. Cho, W. Qiao, F. Tsai, and Y.-H. Lo, “Microfluidics and photonics for Bio-System-on-a-Chip: a review of advancements in technology towards a microfluidic flow cytometry chip,” J. Biophotonics 1(5), 355–376 (2008).
[Crossref] [PubMed]

Tsumura, N.

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

C. Lei, T. Ito, M. Ugawa, T. Nozawa, O. Iwata, M. Maki, G. Okada, H. Kobayashi, X. Sun, P. Tiamsak, N. Tsumura, K. Suzuki, D. Di Carlo, Y. Ozeki, and K. Goda, “High-throughput label-free image cytometry and image-based classification of live Euglena gracilis,” Biomed. Opt. Express 7(7), 2703–2708 (2016).
[Crossref] [PubMed]

Ugawa, M.

van den Engh, G.

G. van den Engh and C. Farmer, “Photo-bleaching and photon saturation in flow cytometry,” Cytometry 13(7), 669–677 (1992).
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van Thourhout, D.

Venkatachalam, V.

D. A. Basiji, W. E. Ortyn, L. Liang, V. Venkatachalam, and P. Morrissey, “Cellular image analysis and imaging by flow cytometry,” Clin. Lab. Med. 27(3), 653–670 (2007).
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Vettenburg, T.

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light-sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
[Crossref] [PubMed]

Viasnoff, V.

R. Galland, G. Grenci, A. Aravind, V. Viasnoff, V. Studer, and J.-B. Sibarita, “3D high- and super-resolution imaging using single-objective SPIM,” Nat. Methods 12(7), 641–644 (2015).
[Crossref] [PubMed]

Wakisaka, Y.

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

Wang, C.

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

Watarai, H.

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

Whitesides, G. M.

S. K. Sia and G. M. Whitesides, “Microfluidic devices fabricated in Poly(dimethylsiloxane) for biological studies,” Electrophoresis 24(21), 3563–3576 (2003).
[Crossref] [PubMed]

Wu, J.

Wu, T.-H.

Y. Chen, T.-H. Wu, Y.-C. Kung, M. A. Teitell, and P.-Y. Chiou, “3D pulsed laser-triggered high-speed microfluidic fluorescence-activated cell sorter,” Analyst 138(24), 7308–7315 (2013).
[Crossref] [PubMed]

Yalikun, Y.

B. Guo, C. Lei, H. Kobayashi, T. Ito, Y. Yalikun, Y. Jiang, Y. Tanaka, Y. Ozeki, and K. Goda, “High-throughput, label-free, single-cell, microalgal lipid screening by machine-learning-equipped optofluidic time-stretch quantitative phase microscopy,” Cytometry A 91(5), 494–502 (2017).
[Crossref] [PubMed]

Yamada, K.

K. Yamada, H. Suzuki, T. Takeuchi, Y. Kazama, S. Mitra, T. Abe, K. Goda, K. Suzuki, and O. Iwata, “Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting,” Sci. Rep. 6(1), 26327 (2016).
[Crossref] [PubMed]

Zagato, E.

Zhang, H.

H. Jiang, T. Zhu, H. Zhang, J. Nie, Z. Guan, C.-M. Ho, S. Liu, and P. Fei, “Droplet-based light-sheet fluorescence microscopy for high-throughput sample preparation, 3-D imaging and quantitative analysis on a chip,” Lab Chip 17(13), 2193–2197 (2017).
[Crossref] [PubMed]

Zhu, T.

H. Jiang, T. Zhu, H. Zhang, J. Nie, Z. Guan, C.-M. Ho, S. Liu, and P. Fei, “Droplet-based light-sheet fluorescence microscopy for high-throughput sample preparation, 3-D imaging and quantitative analysis on a chip,” Lab Chip 17(13), 2193–2197 (2017).
[Crossref] [PubMed]

AIP Adv. (1)

R. Regmi, K. Mohan, and P. P. Mondal, “High resolution light-sheet based high-throughput imaging cytometry system enables visualization of intra-cellular organelles,” AIP Adv. 4(9), 097125 (2014).
[Crossref]

Analyst (1)

Y. Chen, T.-H. Wu, Y.-C. Kung, M. A. Teitell, and P.-Y. Chiou, “3D pulsed laser-triggered high-speed microfluidic fluorescence-activated cell sorter,” Analyst 138(24), 7308–7315 (2013).
[Crossref] [PubMed]

Arch. Mikrobiol. (1)

M. Cramer and J. Myers, “Growth and photosynthetic characteristics of euglena gracilis,” Arch. Mikrobiol. 17(1-4), 384–402 (1952).
[Crossref]

Biomed. Opt. Express (2)

Chem (1)

A. S. Rane, J. Rutkauskaite, A. deMello, and S. Stavrakis, “High-throughput multi-parametric imaging flow cytometry,” Chem 3(4), 588–602 (2017).
[Crossref]

Clin. Lab. Med. (1)

D. A. Basiji, W. E. Ortyn, L. Liang, V. Venkatachalam, and P. Morrissey, “Cellular image analysis and imaging by flow cytometry,” Clin. Lab. Med. 27(3), 653–670 (2007).
[Crossref] [PubMed]

Cytometry (1)

G. van den Engh and C. Farmer, “Photo-bleaching and photon saturation in flow cytometry,” Cytometry 13(7), 669–677 (1992).
[Crossref] [PubMed]

Cytometry A (1)

B. Guo, C. Lei, H. Kobayashi, T. Ito, Y. Yalikun, Y. Jiang, Y. Tanaka, Y. Ozeki, and K. Goda, “High-throughput, label-free, single-cell, microalgal lipid screening by machine-learning-equipped optofluidic time-stretch quantitative phase microscopy,” Cytometry A 91(5), 494–502 (2017).
[Crossref] [PubMed]

Electrophoresis (1)

S. K. Sia and G. M. Whitesides, “Microfluidic devices fabricated in Poly(dimethylsiloxane) for biological studies,” Electrophoresis 24(21), 3563–3576 (2003).
[Crossref] [PubMed]

J. Biophotonics (1)

J. Godin, C.-H. Chen, S. H. Cho, W. Qiao, F. Tsai, and Y.-H. Lo, “Microfluidics and photonics for Bio-System-on-a-Chip: a review of advancements in technology towards a microfluidic flow cytometry chip,” J. Biophotonics 1(5), 355–376 (2008).
[Crossref] [PubMed]

Lab Chip (3)

H. Jiang, T. Zhu, H. Zhang, J. Nie, Z. Guan, C.-M. Ho, S. Liu, and P. Fei, “Droplet-based light-sheet fluorescence microscopy for high-throughput sample preparation, 3-D imaging and quantitative analysis on a chip,” Lab Chip 17(13), 2193–2197 (2017).
[Crossref] [PubMed]

P. Paiè, F. Bragheri, A. Bassi, and R. Osellame, “Selective plane illumination microscopy on a chip,” Lab Chip 16(9), 1556–1560 (2016).
[Crossref] [PubMed]

M. Nordin and T. Laurell, “Two-hundredfold volume concentration of dilute cell and particle suspensions using chip integrated multistage acoustophoresis,” Lab Chip 12(22), 4610–4616 (2012).
[Crossref] [PubMed]

Microsc. Res. Tech. (1)

R. Regmi, K. Mohan, and P. P. Mondal, “MRT letter: light sheet based imaging flow cytometry on a microfluidic platform,” Microsc. Res. Tech. 76(11), 1101–1107 (2013).
[Crossref] [PubMed]

Nat. Methods (4)

T. Vettenburg, H. I. C. Dalgarno, J. Nylk, C. Coll-Lladó, D. E. K. Ferrier, T. Čižmár, F. J. Gunn-Moore, and K. Dholakia, “Light-sheet microscopy using an Airy beam,” Nat. Methods 11(5), 541–544 (2014).
[Crossref] [PubMed]

T. A. Planchon, L. Gao, D. E. Milkie, M. W. Davidson, J. A. Galbraith, C. G. Galbraith, and E. Betzig, “Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination,” Nat. Methods 8(5), 417–423 (2011).
[Crossref] [PubMed]

R. M. Power and J. Huisken, “A guide to light-sheet fluorescence microscopy for multiscale imaging,” Nat. Methods 14(4), 360–373 (2017).
[Crossref] [PubMed]

R. Galland, G. Grenci, A. Aravind, V. Viasnoff, V. Studer, and J.-B. Sibarita, “3D high- and super-resolution imaging using single-objective SPIM,” Nat. Methods 12(7), 641–644 (2015).
[Crossref] [PubMed]

Nat. Microbiol. (1)

Y. Wakisaka, Y. Suzuki, O. Iwata, A. Nakashima, T. Ito, M. Hirose, R. Domon, M. Sugawara, N. Tsumura, H. Watarai, T. Shimobaba, K. Suzuki, K. Goda, and Y. Ozeki, “Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy,” Nat. Microbiol. 1(10), 16124 (2016).
[Crossref] [PubMed]

Nature (1)

J. El-Ali, P. K. Sorger, and K. F. Jensen, “Cells on chips,” Nature 442, 403–411 (2006).
[Crossref] [PubMed]

Opt. Express (3)

Plant Sci. Lett. (1)

R. Schantz, M.-L. Schantz, and H. Duranton, “Changes in amino acid and peptide composition of Euglena gracilis cells during chloroplast development,” Plant Sci. Lett. 5(5), 313–324 (1975).
[Crossref]

PLoS One (1)

B. Guo, C. Lei, T. Ito, Y. Jiang, Y. Ozeki, and K. Goda, “High-throughput accurate single-cell screening of Euglena gracilis with fluorescence-assisted optofluidic time-stretch microscopy,” PLoS One 11(11), e0166214 (2016).
[Crossref] [PubMed]

Proc. Natl. Acad. Sci. U.S.A. (1)

K. Goda, A. Ayazi, D. R. Gossett, J. Sadasivam, C. K. Lonappan, E. Sollier, A. M. Fard, S. C. Hur, J. Adam, C. Murray, C. Wang, N. Brackbill, D. Di Carlo, and B. Jalali, “High-throughput single-microparticle imaging flow analyzer,” Proc. Natl. Acad. Sci. U.S.A. 109(29), 11630–11635 (2012).
[Crossref] [PubMed]

Sci. Rep. (1)

K. Yamada, H. Suzuki, T. Takeuchi, Y. Kazama, S. Mitra, T. Abe, K. Goda, K. Suzuki, and O. Iwata, “Efficient selective breeding of live oil-rich Euglena gracilis with fluorescence-activated cell sorting,” Sci. Rep. 6(1), 26327 (2016).
[Crossref] [PubMed]

Other (5)

Brochure of ImageStreamX, http://www.emdmillipore.com/US/en/life-science-research/cell-analysis-flow-cytometry/amnis-imaging-flow-cytometers/imagestreamx-Mark-ii-imaging-flow-cytometer/VaSb.qB.QokAAAFLzRop.zHe,nav?bd=1

N. S. Barteneva and I. A. Vorobjev, Imaging Flow Cytometry (Springer, 2016).

M. M. Watanabe, M. Kawachi, M. Hiroki, F. Kasai, and Eds, NIES Collection List of Strains. Ed. 6, NIES, Japan (2000).

L. Wang, ed., Support Vector Machines: Theory and Applications (Springer, 2005).

N. Cristianini and J. Shawe-Taylor, An Introduction to Support Vector Machines and Other Kernel-based Learning Methods (Cambridge University Press, 2000).

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Figures (6)

Fig. 1
Fig. 1 Principles of light-sheet excitation in comparison with wide-field excitation. (a) Schematic of the light-sheet excitation of flowing cells in the microchannel on the mirror-embedded microfluidic chip. (b) Schematic of the wide-field excitation of cells flowing in a microchannel on a standard microfluidic chip. (c) Schematic of the cross section of the light-sheet excitation beam. (d) Schematic of the cross section of the wide-field excitation beam.
Fig. 2
Fig. 2 Schematic of the high-throughput on-chip light-sheet fluorescence imaging flow cytometer. FG: function generator; TL: tube lens; DM: dichroic mirror; M: mirror; SL: spherical lens; CL: cylindrical lens; OL: objective lens; Inlet 1: sample flow inlet; Inlet 2: sheath flow inlet.
Fig. 3
Fig. 3 Mirror-embedded microfluidic chip. (a) Schematic of the microchannel. (i) and (ii) show the hydrodynamic focusing and optical interrogation regions, respectively. (b) Fabrication process. (c) Picture of the fabricated microfluidic chip. (d) Zoomed picture of the microchannel and aluminum-coated glass, indicating the precise alignment of the aluminum-coated glass to the microchannel.
Fig. 4
Fig. 4 Evaluation of the on-chip fluorescence imaging flow cytometer. (a) Fluorescence images of a FITC solution under the wide-field excitation (left) and light-sheet excitation (right). (b) Bright-field image of static E. gracilis cells. (c) Acquired fluorescence (autofluorescence from intracellular chlorophyll) images of a single E. gracilis cell and profiles of the fluorescence intensity along the white dashed lines in the images under the wide-field excitation (left) and light-sheet excitation (right).
Fig. 5
Fig. 5 Image-based classification of E. gracilis cells under different culture conditions. (a) Obtained image library of the cells. Top: cells cultivated under a nitrogen-sufficient condition. Bottom: cells cultivated under a nitrogen-deficient condition. Green: intracellular lipids stained by BODIPY505/515. Red: intracellular chlorophyll (autofluorescence). (b) Scatter plot of the cells (N = 3,000 for each culture). (c) Histogram of the cells with a meta-feature obtained by a support vector machine.
Fig. 6
Fig. 6 Image-based classification of MCF-7 (human breast adenocarcinoma) and HT-29 (human colorectal adenocarcinoma) cells. (a) Obtained images of the cells. Top: MCF-7 cells. Bottom: HT-29 cells. Green: nucleus stained by SYTO16. Red: cytoplasm stained by CellTracker Red. (b) Scatter plot of the cells (N = 2,800 for each culture). (c) Histogram of the cells with a meta-feature obtained by a support vector machine.

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