Abstract

Coherent anti-stokes Raman scattering (CARS) flow cytometry was demonstrated by combining a laser-scanning CARS microscope with a polydimethylsiloxane (PDMS) based microfluidic device. Line-scanning across the hydrodynamically focused core stream was performed for detection of flowing objects. Parameters were optimized by utilizing polystyrene beads as flowing particles. Population measurements of adipocytes isolated from mouse fat tissues demonstrated the viability of microfluidic CARS cytometry for quantitation of adipocyte size distribution. CARS cytometry could be a new modality for quantitative analysis with vibrational selectivity.

© 2008 Optical Society of America

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References

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    [Crossref]
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    [Crossref]
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    [Crossref]
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    [Crossref] [PubMed]
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    [Crossref] [PubMed]

2007 (7)

M. Muller and A. Zumbusch, “Coherent anti-Stokes Raman Scattering Microscopy,” Chem. PhysChem. 8, 2156–2170 (2007).
[Crossref]

T. D. Chung and H. C. Kim, “Recent advances in miniaturized microfluidic flow cytometry for clinical use,” Electrophoresis 28, 4511–4520 (2007).
[Crossref] [PubMed]

A. Gordon, A. Colman-Lerner, T. E. Chin, K. R. Benjamin, R. C. Yu, and R. Brent, “Single-cell quantification of molecules and rates using open-source microscope-based cytometry,” Nat. Methods 4, 175–181 (2007).
[Crossref] [PubMed]

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[Crossref] [PubMed]

Y. Fu, H. Wang, R. Shi, and J. X. Cheng, “Second harmonic and sum frequency generation imaging of fibrous astroglial filaments in ex vivo spinal tissues,” Biophys. J. 92, 3251–3259 (2007).
[Crossref] [PubMed]

K. Bahlmann, P. T. So, M. Kirber, R. Reich, B. Kosicki, W. McGonagle, and K. Bellve, “Multifocal multiphoton microscopy (MMM) at a frame rate beyond 600 Hz,” Opt. Express 15, 10991–10998 (2007).
[Crossref] [PubMed]

J. X. Cheng, “Coherent anti-stokes Raman scattering microscopy,” Appl. Spectrosc. 61, 197A–208A (2007).
[Crossref]

2006 (1)

H. Y. Wang and C. Lu, “Electroporation of Mammalian Cells in a Microfluidic Channel with Geometric Variation,” Anal. Chem. 78, 5158–5164 (2006).
[Crossref] [PubMed]

2005 (2)

H. Wang, Y. Fu, P. Zickmund, R. Shi, and J. X. Cheng, “Coherent anti-stokes Raman scattering imaging of axonl myelin in live spinal tissues.,” Biophys. J. 89, 581–591 (2005).
[Crossref] [PubMed]

D. Huh, W. Gu, Y. Kamotani, J. B. Grotberg, and S. Takayama, “Microfluidics for flow cytometric analysis of cells and particles,” Physiol. Meas. 26, R73–R98 (2005).
[Crossref] [PubMed]

2004 (2)

S. P. Perfetto, P. K. Chattopadhyay, and M. Roederer, “Seventeen-colour flow cytometry: unravelling the immune system,” Nat. Rev. Immun. 4, 648–655 (2004).
[Crossref]

J. X. Cheng and X. S. Xie, “Coherent anti-Stokes Raman scattering microscopy: instrumentation, theory, and applications.,” J. Phys. Chem. B 108, 827–840 (2004).
[Crossref]

2001 (1)

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “An epi-detected coherent anti-stokes Raman scattering microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
[Crossref]

1999 (2)

K. H. Kim, C. Buehler, and P. T. C. So, “High-Speed, Two-Photon Scanning Microscope,” Appl. Opt. 38, 6004–6009 (1999).
[Crossref]

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

1998 (1)

D. C. Duffy, J. C. McDonald, O. J. A. Schueller, and G. M. Whitesides, “Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane),” Anal. Chem. 70, 4974–4984 (1998).
[Crossref] [PubMed]

1997 (1)

C. D. Jennings and K. A. Foon, “Recent Advances in Flow Cytometry: Application to the Diagnosis of Hematologic Malignancy,” Blood 90, 2863–2892 (1997).
[PubMed]

Bahlmann, K.

Bellve, K.

Benjamin, K. R.

A. Gordon, A. Colman-Lerner, T. E. Chin, K. R. Benjamin, R. C. Yu, and R. Brent, “Single-cell quantification of molecules and rates using open-source microscope-based cytometry,” Nat. Methods 4, 175–181 (2007).
[Crossref] [PubMed]

Book, L. D.

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “An epi-detected coherent anti-stokes Raman scattering microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
[Crossref]

Brent, R.

A. Gordon, A. Colman-Lerner, T. E. Chin, K. R. Benjamin, R. C. Yu, and R. Brent, “Single-cell quantification of molecules and rates using open-source microscope-based cytometry,” Nat. Methods 4, 175–181 (2007).
[Crossref] [PubMed]

Buehler, C.

Chattopadhyay, P. K.

S. P. Perfetto, P. K. Chattopadhyay, and M. Roederer, “Seventeen-colour flow cytometry: unravelling the immune system,” Nat. Rev. Immun. 4, 648–655 (2004).
[Crossref]

Cheng, J. X.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[Crossref] [PubMed]

Y. Fu, H. Wang, R. Shi, and J. X. Cheng, “Second harmonic and sum frequency generation imaging of fibrous astroglial filaments in ex vivo spinal tissues,” Biophys. J. 92, 3251–3259 (2007).
[Crossref] [PubMed]

J. X. Cheng, “Coherent anti-stokes Raman scattering microscopy,” Appl. Spectrosc. 61, 197A–208A (2007).
[Crossref]

H. Wang, Y. Fu, P. Zickmund, R. Shi, and J. X. Cheng, “Coherent anti-stokes Raman scattering imaging of axonl myelin in live spinal tissues.,” Biophys. J. 89, 581–591 (2005).
[Crossref] [PubMed]

J. X. Cheng and X. S. Xie, “Coherent anti-Stokes Raman scattering microscopy: instrumentation, theory, and applications.,” J. Phys. Chem. B 108, 827–840 (2004).
[Crossref]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “An epi-detected coherent anti-stokes Raman scattering microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
[Crossref]

Chin, T. E.

A. Gordon, A. Colman-Lerner, T. E. Chin, K. R. Benjamin, R. C. Yu, and R. Brent, “Single-cell quantification of molecules and rates using open-source microscope-based cytometry,” Nat. Methods 4, 175–181 (2007).
[Crossref] [PubMed]

Chung, T. D.

T. D. Chung and H. C. Kim, “Recent advances in miniaturized microfluidic flow cytometry for clinical use,” Electrophoresis 28, 4511–4520 (2007).
[Crossref] [PubMed]

Colman-Lerner, A.

A. Gordon, A. Colman-Lerner, T. E. Chin, K. R. Benjamin, R. C. Yu, and R. Brent, “Single-cell quantification of molecules and rates using open-source microscope-based cytometry,” Nat. Methods 4, 175–181 (2007).
[Crossref] [PubMed]

Duffy, D. C.

D. C. Duffy, J. C. McDonald, O. J. A. Schueller, and G. M. Whitesides, “Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane),” Anal. Chem. 70, 4974–4984 (1998).
[Crossref] [PubMed]

Foon, K. A.

C. D. Jennings and K. A. Foon, “Recent Advances in Flow Cytometry: Application to the Diagnosis of Hematologic Malignancy,” Blood 90, 2863–2892 (1997).
[PubMed]

Fu, Y.

Y. Fu, H. Wang, R. Shi, and J. X. Cheng, “Second harmonic and sum frequency generation imaging of fibrous astroglial filaments in ex vivo spinal tissues,” Biophys. J. 92, 3251–3259 (2007).
[Crossref] [PubMed]

H. Wang, Y. Fu, P. Zickmund, R. Shi, and J. X. Cheng, “Coherent anti-stokes Raman scattering imaging of axonl myelin in live spinal tissues.,” Biophys. J. 89, 581–591 (2005).
[Crossref] [PubMed]

Gordon, A.

A. Gordon, A. Colman-Lerner, T. E. Chin, K. R. Benjamin, R. C. Yu, and R. Brent, “Single-cell quantification of molecules and rates using open-source microscope-based cytometry,” Nat. Methods 4, 175–181 (2007).
[Crossref] [PubMed]

Grotberg, J. B.

D. Huh, W. Gu, Y. Kamotani, J. B. Grotberg, and S. Takayama, “Microfluidics for flow cytometric analysis of cells and particles,” Physiol. Meas. 26, R73–R98 (2005).
[Crossref] [PubMed]

Gu, W.

D. Huh, W. Gu, Y. Kamotani, J. B. Grotberg, and S. Takayama, “Microfluidics for flow cytometric analysis of cells and particles,” Physiol. Meas. 26, R73–R98 (2005).
[Crossref] [PubMed]

Holtom, G. R.

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

Huh, D.

D. Huh, W. Gu, Y. Kamotani, J. B. Grotberg, and S. Takayama, “Microfluidics for flow cytometric analysis of cells and particles,” Physiol. Meas. 26, R73–R98 (2005).
[Crossref] [PubMed]

Jennings, C. D.

C. D. Jennings and K. A. Foon, “Recent Advances in Flow Cytometry: Application to the Diagnosis of Hematologic Malignancy,” Blood 90, 2863–2892 (1997).
[PubMed]

Kamotani, Y.

D. Huh, W. Gu, Y. Kamotani, J. B. Grotberg, and S. Takayama, “Microfluidics for flow cytometric analysis of cells and particles,” Physiol. Meas. 26, R73–R98 (2005).
[Crossref] [PubMed]

Kim, H. C.

T. D. Chung and H. C. Kim, “Recent advances in miniaturized microfluidic flow cytometry for clinical use,” Electrophoresis 28, 4511–4520 (2007).
[Crossref] [PubMed]

Kim, K. H.

Kirber, M.

Kosicki, B.

Langohr, I. M.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[Crossref] [PubMed]

Le, T. T.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[Crossref] [PubMed]

Locker, M. J.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[Crossref] [PubMed]

Lu, C.

H. Y. Wang and C. Lu, “Electroporation of Mammalian Cells in a Microfluidic Channel with Geometric Variation,” Anal. Chem. 78, 5158–5164 (2006).
[Crossref] [PubMed]

H.-Y. Wang and C. Lu, “Microfluidic chemical cytometry based on modulation of local field strength,” Chem. Commun.3528–3530 (2006).
[Crossref]

McDonald, J. C.

D. C. Duffy, J. C. McDonald, O. J. A. Schueller, and G. M. Whitesides, “Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane),” Anal. Chem. 70, 4974–4984 (1998).
[Crossref] [PubMed]

McGonagle, W.

Muller, M.

M. Muller and A. Zumbusch, “Coherent anti-Stokes Raman Scattering Microscopy,” Chem. PhysChem. 8, 2156–2170 (2007).
[Crossref]

Perfetto, S. P.

S. P. Perfetto, P. K. Chattopadhyay, and M. Roederer, “Seventeen-colour flow cytometry: unravelling the immune system,” Nat. Rev. Immun. 4, 648–655 (2004).
[Crossref]

Reich, R.

Roederer, M.

S. P. Perfetto, P. K. Chattopadhyay, and M. Roederer, “Seventeen-colour flow cytometry: unravelling the immune system,” Nat. Rev. Immun. 4, 648–655 (2004).
[Crossref]

Schueller, O. J. A.

D. C. Duffy, J. C. McDonald, O. J. A. Schueller, and G. M. Whitesides, “Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane),” Anal. Chem. 70, 4974–4984 (1998).
[Crossref] [PubMed]

Shapiro, H. M.

H. M. Shapiro, Practical Flow Cytometry, 4th ed. (Wiley Liss, New York, 2003).
[Crossref]

Shi, R.

Y. Fu, H. Wang, R. Shi, and J. X. Cheng, “Second harmonic and sum frequency generation imaging of fibrous astroglial filaments in ex vivo spinal tissues,” Biophys. J. 92, 3251–3259 (2007).
[Crossref] [PubMed]

H. Wang, Y. Fu, P. Zickmund, R. Shi, and J. X. Cheng, “Coherent anti-stokes Raman scattering imaging of axonl myelin in live spinal tissues.,” Biophys. J. 89, 581–591 (2005).
[Crossref] [PubMed]

So, P. T.

So, P. T. C.

Sturek, M.

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[Crossref] [PubMed]

Takayama, S.

D. Huh, W. Gu, Y. Kamotani, J. B. Grotberg, and S. Takayama, “Microfluidics for flow cytometric analysis of cells and particles,” Physiol. Meas. 26, R73–R98 (2005).
[Crossref] [PubMed]

Volkmer, A.

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “An epi-detected coherent anti-stokes Raman scattering microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
[Crossref]

Wang, H.

Y. Fu, H. Wang, R. Shi, and J. X. Cheng, “Second harmonic and sum frequency generation imaging of fibrous astroglial filaments in ex vivo spinal tissues,” Biophys. J. 92, 3251–3259 (2007).
[Crossref] [PubMed]

H. Wang, Y. Fu, P. Zickmund, R. Shi, and J. X. Cheng, “Coherent anti-stokes Raman scattering imaging of axonl myelin in live spinal tissues.,” Biophys. J. 89, 581–591 (2005).
[Crossref] [PubMed]

Wang, H. Y.

H. Y. Wang and C. Lu, “Electroporation of Mammalian Cells in a Microfluidic Channel with Geometric Variation,” Anal. Chem. 78, 5158–5164 (2006).
[Crossref] [PubMed]

Wang, H.-Y.

H.-Y. Wang and C. Lu, “Microfluidic chemical cytometry based on modulation of local field strength,” Chem. Commun.3528–3530 (2006).
[Crossref]

Whitesides, G. M.

D. C. Duffy, J. C. McDonald, O. J. A. Schueller, and G. M. Whitesides, “Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane),” Anal. Chem. 70, 4974–4984 (1998).
[Crossref] [PubMed]

Xie, X. S.

J. X. Cheng and X. S. Xie, “Coherent anti-Stokes Raman scattering microscopy: instrumentation, theory, and applications.,” J. Phys. Chem. B 108, 827–840 (2004).
[Crossref]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “An epi-detected coherent anti-stokes Raman scattering microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
[Crossref]

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

Yu, R. C.

A. Gordon, A. Colman-Lerner, T. E. Chin, K. R. Benjamin, R. C. Yu, and R. Brent, “Single-cell quantification of molecules and rates using open-source microscope-based cytometry,” Nat. Methods 4, 175–181 (2007).
[Crossref] [PubMed]

Zickmund, P.

H. Wang, Y. Fu, P. Zickmund, R. Shi, and J. X. Cheng, “Coherent anti-stokes Raman scattering imaging of axonl myelin in live spinal tissues.,” Biophys. J. 89, 581–591 (2005).
[Crossref] [PubMed]

Zumbusch, A.

M. Muller and A. Zumbusch, “Coherent anti-Stokes Raman Scattering Microscopy,” Chem. PhysChem. 8, 2156–2170 (2007).
[Crossref]

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

Anal. Chem. (2)

H. Y. Wang and C. Lu, “Electroporation of Mammalian Cells in a Microfluidic Channel with Geometric Variation,” Anal. Chem. 78, 5158–5164 (2006).
[Crossref] [PubMed]

D. C. Duffy, J. C. McDonald, O. J. A. Schueller, and G. M. Whitesides, “Rapid Prototyping of Microfluidic Systems in Poly(dimethylsiloxane),” Anal. Chem. 70, 4974–4984 (1998).
[Crossref] [PubMed]

Appl. Opt. (1)

Appl. Spectrosc. (1)

Biophys. J. (2)

H. Wang, Y. Fu, P. Zickmund, R. Shi, and J. X. Cheng, “Coherent anti-stokes Raman scattering imaging of axonl myelin in live spinal tissues.,” Biophys. J. 89, 581–591 (2005).
[Crossref] [PubMed]

Y. Fu, H. Wang, R. Shi, and J. X. Cheng, “Second harmonic and sum frequency generation imaging of fibrous astroglial filaments in ex vivo spinal tissues,” Biophys. J. 92, 3251–3259 (2007).
[Crossref] [PubMed]

Blood (1)

C. D. Jennings and K. A. Foon, “Recent Advances in Flow Cytometry: Application to the Diagnosis of Hematologic Malignancy,” Blood 90, 2863–2892 (1997).
[PubMed]

Chem. PhysChem. (1)

M. Muller and A. Zumbusch, “Coherent anti-Stokes Raman Scattering Microscopy,” Chem. PhysChem. 8, 2156–2170 (2007).
[Crossref]

Electrophoresis (1)

T. D. Chung and H. C. Kim, “Recent advances in miniaturized microfluidic flow cytometry for clinical use,” Electrophoresis 28, 4511–4520 (2007).
[Crossref] [PubMed]

J. Biomed. Opt. (1)

T. T. Le, I. M. Langohr, M. J. Locker, M. Sturek, and J. X. Cheng, “Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy,” J. Biomed. Opt. 12, 054007 (2007).
[Crossref] [PubMed]

J. Phys. Chem. B (2)

J. X. Cheng and X. S. Xie, “Coherent anti-Stokes Raman scattering microscopy: instrumentation, theory, and applications.,” J. Phys. Chem. B 108, 827–840 (2004).
[Crossref]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, “An epi-detected coherent anti-stokes Raman scattering microscope with high spectral resolution and high sensitivity,” J. Phys. Chem. B 105, 1277–1280 (2001).
[Crossref]

Nat. Methods (1)

A. Gordon, A. Colman-Lerner, T. E. Chin, K. R. Benjamin, R. C. Yu, and R. Brent, “Single-cell quantification of molecules and rates using open-source microscope-based cytometry,” Nat. Methods 4, 175–181 (2007).
[Crossref] [PubMed]

Nat. Rev. Immun. (1)

S. P. Perfetto, P. K. Chattopadhyay, and M. Roederer, “Seventeen-colour flow cytometry: unravelling the immune system,” Nat. Rev. Immun. 4, 648–655 (2004).
[Crossref]

Opt. Express (1)

Phys. Rev. Lett. (1)

A. Zumbusch, G. R. Holtom, and X. S. Xie, “Three-dimensional vibrational imaging by coherent anti-Stokes Raman scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

Physiol. Meas. (1)

D. Huh, W. Gu, Y. Kamotani, J. B. Grotberg, and S. Takayama, “Microfluidics for flow cytometric analysis of cells and particles,” Physiol. Meas. 26, R73–R98 (2005).
[Crossref] [PubMed]

Other (2)

H. M. Shapiro, Practical Flow Cytometry, 4th ed. (Wiley Liss, New York, 2003).
[Crossref]

H.-Y. Wang and C. Lu, “Microfluidic chemical cytometry based on modulation of local field strength,” Chem. Commun.3528–3530 (2006).
[Crossref]

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Figures (4)

Fig. 1.
Fig. 1.

Schematic diagram of CARS cytometry and imaging illustration of the hydrodynamic focusing effect. (a) Schematic of microfluidic CARS cytometry. The blowup inset shows layers of microfluidic chip providing function of hydrodynamic focusing for cytometry analysis. The schematic drawings are not to scale. D.M=dichroic mirror; Obj=objective. (b) Hydrodynamic focusing effect examined by forward CARS imaging of D2O injected as the core flow (red) from inlet A, and backward TPEF imaging of fluorescent microspheres in the sheath flow (green) injected from inlet B. The bar over the core flow indicates the line-scan window. (c) CARS spectrum of D2O measured by tuning the frequency of the slave laser from 11300 to 12085 cm-1. The peak is located around 2350 cm-1. (d) Transmission image showing 2 µm beads in the core around the flow impinging area in a microfluidic chip. Arrows show the flow directions.

Fig. 2.
Fig. 2.

Variations of bead size and signal intensity correlated to S/F ratio. (a) S/F ratio versus mean size of 10 µm PS beads. As increasing the S/F ratio, size variation was reduced and the mean size was closer to the true value. (b) S/F ratio versus mean size of 2 µm PS beads. The size variation mainly resulted from the unmatched channel depth. The insets tagged A, B, and C in (a) and (b) are the time-stack CARS images (~160 ms) relative to the S/F ratio shown in the plots. Time-stack CARS images were obtained by signaling along the fixed 1-D scan window of ~34 µm. Arrows besides inset C indicate the direction of time flow. (c) S/F ratio versus mean signal intensity of 10 µm PS beads. (d) Signal history of 10 µm PS beads within 1 ms. The signal trace was obtained by setting the S/F ratio to be 0.203. All CARS signals were backward detected.

Fig. 3.
Fig. 3.

Population distribution of size mixed PS beads verified by microfluidic CARS cytometry. (a) A typical time-stack CARS image (~600 ms) obtained by signaling along the 1-D scan window of ~34 µm for population analysis. The arrow indicates the direction of time flow. (b) Bead counts versus size of a solution mixed with 5 µm and 10 µm beads. The S/F ratio was set ~0.2.

Fig. 4.
Fig. 4.

Size population of adipocytes verified by microfluidic CARS cytometry. (a) Size distribution of isolated adipocytes retrieved from time stack images of CARS cytometry. The S/F ratio was set ~0.2. (b) Size distribution of isolated adipocytes obtained by analyzing 8 CARS images for microscopic-based cytometry. Size analyses were obtained by using a program developed on MATLAB platform. The inset in (b) shows a backward-detected CARS image of purified adipocytes from mouse fat tissues. Bar=30 µm.

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