Abstract

We have developed fully motorized optical-resolution photoacoustic microscopy (OR-PAM), which integrates five complementary scanning modes and simultaneously provides a high imaging speed and a wide field of view (FOV) with 2.6 μm lateral resolution. With one-dimensional (1D) motion-mode mechanical scanning, we measured the blood flow through a cross section of a blood vessel in vivo. With two-dimensional (2D) optical scanning at a laser repetition rate of 40 kHz, we achieved a 2 kHz B-scan rate over a range of 50 μm with 20 A-lines and 50 Hz volumetric-scan rate over a FOV of 50μm×50μm with 400 A-lines, which enabled real-time tracking of cellular dynamics in vivo. With synchronized 1D optical and 2D mechanical hybrid scanning, we imaged a 10mm×8mm FOV within three minutes, which is 20 times faster than the conventional mechanical scan in our second-generation OR-PAM. With three-dimensional mechanical contour scanning, we maintained the optimal signal-to-noise ratio and spatial resolution of OR-PAM while imaging objects with uneven surfaces, which is essential for quantitative studies.

© 2014 Optical Society of America

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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
  6. R. L. Shelton, S. P. Mattison, and B. E. Applegate, “Volumetric imaging of erythrocytes using label-free multiphoton photoacoustic microscopy,” J. Biophotonics, doi: 10.1002/jbio.201300059.
    [CrossRef]
  7. R. J. Paproski, A. E. Forbrich, K. Wachowicz, M. M. Hitt, and R. J. Zemp, Biomed. Opt. Express 2, 771 (2011).
    [CrossRef]
  8. G. Suffredini, J. E. East, and L. M. Levy, “New applications of nanotechnology for neuroimaging,” Am. J. Neuroradiol. (to be published).
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
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  17. J. J. Yao, K. I. Maslov, Y. Zhang, Y. N. Xia, and L. V. Wang, J. Biomed. Opt. 16, 076003 (2011).
    [CrossRef]
  18. H. F. Zhang, K. Maslov, M. L. Li, G. Stoica, and L. H. V. Wang, Opt. Express 14, 9317 (2006).
    [CrossRef]
  19. L. D. Wang, K. Maslov, and L. H. V. Wang, Proc. Natl. Acad. Sci. USA 110, 5759 (2013).
    [CrossRef]
  20. P. Hajireza, W. Shi, and R. J. Zemp, Opt. Lett. 36, 4107 (2011).
    [CrossRef]

2013

T. Liu, H. Li, W. Song, S. L. Jiao, and H. F. Zhang, Curr. Eye Res. 38, 1229 (2013).
[CrossRef]

W. Song, Q. Wei, S. Jiao, and H. F. Zhang, J. Visualized Exp. 71, e4390 (2013).

L. D. Wang, K. Maslov, and L. H. V. Wang, Proc. Natl. Acad. Sci. USA 110, 5759 (2013).
[CrossRef]

2012

L. V. Wang and S. Hu, Science 335, 1458 (2012).
[CrossRef]

Y. Jiang, A. Forbrich, T. Harrison, and R. J. Zemp, J. Biomed. Opt. 17, 036012 (2012).
[CrossRef]

A. Ray, J. R. Rajian, Y. E. Lee, X. Wang, and R. Kopelman, J. Biomed. Opt. 17, 057004 (2012).
[CrossRef]

2011

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

S. Hu, K. Maslov, and L. V. Wang, Opt. Lett. 36, 269 (2011).

J. J. Yao, K. I. Maslov, Y. Zhang, Y. N. Xia, and L. V. Wang, J. Biomed. Opt. 16, 076003 (2011).
[CrossRef]

R. J. Paproski, A. E. Forbrich, K. Wachowicz, M. M. Hitt, and R. J. Zemp, Biomed. Opt. Express 2, 771 (2011).
[CrossRef]

P. Hajireza, W. Shi, and R. J. Zemp, Opt. Lett. 36, 4107 (2011).
[CrossRef]

2010

2008

2006

Applegate, B. E.

R. L. Shelton and B. E. Applegate, Biomed. Opt. Express 1, 676 (2010).
[CrossRef]

R. L. Shelton, S. P. Mattison, and B. E. Applegate, “Volumetric imaging of erythrocytes using label-free multiphoton photoacoustic microscopy,” J. Biophotonics, doi: 10.1002/jbio.201300059.
[CrossRef]

Arbeit, J. M.

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

East, J. E.

G. Suffredini, J. E. East, and L. M. Levy, “New applications of nanotechnology for neuroimaging,” Am. J. Neuroradiol. (to be published).

Fawzi, A.

Forbrich, A.

Y. Jiang, A. Forbrich, T. Harrison, and R. J. Zemp, J. Biomed. Opt. 17, 036012 (2012).
[CrossRef]

Forbrich, A. E.

Hajireza, P.

Harrison, T.

Y. Jiang, A. Forbrich, T. Harrison, and R. J. Zemp, J. Biomed. Opt. 17, 036012 (2012).
[CrossRef]

Hitt, M. M.

Hu, J. M.

Hu, S.

L. V. Wang and S. Hu, Science 335, 1458 (2012).
[CrossRef]

S. Hu, K. Maslov, and L. V. Wang, Opt. Lett. 36, 269 (2011).

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

S. Hu, B. Rao, K. Maslov, and L. V. Wang, Opt. Lett. 35, 1 (2010).
[CrossRef]

S. Hu and L. V. Wang, Front. Neuroenerg. 2, 10 (2010).

K. Maslov, H. F. Zhang, S. Hu, and L. V. Wang, Opt. Lett. 33, 929 (2008).
[CrossRef]

Jiang, M. S.

Jiang, Y.

Y. Jiang, A. Forbrich, T. Harrison, and R. J. Zemp, J. Biomed. Opt. 17, 036012 (2012).
[CrossRef]

Jiao, S.

W. Song, Q. Wei, S. Jiao, and H. F. Zhang, J. Visualized Exp. 71, e4390 (2013).

Jiao, S. L.

Kopelman, R.

A. Ray, J. R. Rajian, Y. E. Lee, X. Wang, and R. Kopelman, J. Biomed. Opt. 17, 057004 (2012).
[CrossRef]

Kovalski, J. R.

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

Lee, Y. E.

A. Ray, J. R. Rajian, Y. E. Lee, X. Wang, and R. Kopelman, J. Biomed. Opt. 17, 057004 (2012).
[CrossRef]

Levy, L. M.

G. Suffredini, J. E. East, and L. M. Levy, “New applications of nanotechnology for neuroimaging,” Am. J. Neuroradiol. (to be published).

Li, H.

T. Liu, H. Li, W. Song, S. L. Jiao, and H. F. Zhang, Curr. Eye Res. 38, 1229 (2013).
[CrossRef]

Li, M. L.

Liu, T.

T. Liu, H. Li, W. Song, S. L. Jiao, and H. F. Zhang, Curr. Eye Res. 38, 1229 (2013).
[CrossRef]

Maslov, K.

L. D. Wang, K. Maslov, and L. H. V. Wang, Proc. Natl. Acad. Sci. USA 110, 5759 (2013).
[CrossRef]

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

S. Hu, K. Maslov, and L. V. Wang, Opt. Lett. 36, 269 (2011).

S. Hu, B. Rao, K. Maslov, and L. V. Wang, Opt. Lett. 35, 1 (2010).
[CrossRef]

K. Maslov, H. F. Zhang, S. Hu, and L. V. Wang, Opt. Lett. 33, 929 (2008).
[CrossRef]

H. F. Zhang, K. Maslov, M. L. Li, G. Stoica, and L. H. V. Wang, Opt. Express 14, 9317 (2006).
[CrossRef]

Maslov, K. I.

J. J. Yao, K. I. Maslov, Y. Zhang, Y. N. Xia, and L. V. Wang, J. Biomed. Opt. 16, 076003 (2011).
[CrossRef]

J. J. Yao, K. I. Maslov, Y. F. Shi, L. A. Taber, and L. H. V. Wang, Opt. Lett. 35, 1419 (2010).
[CrossRef]

Mattison, S. P.

R. L. Shelton, S. P. Mattison, and B. E. Applegate, “Volumetric imaging of erythrocytes using label-free multiphoton photoacoustic microscopy,” J. Biophotonics, doi: 10.1002/jbio.201300059.
[CrossRef]

Oladipupo, S. S.

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

Paproski, R. J.

Puliafito, C. A.

Rajian, J. R.

A. Ray, J. R. Rajian, Y. E. Lee, X. Wang, and R. Kopelman, J. Biomed. Opt. 17, 057004 (2012).
[CrossRef]

Rao, B.

Ray, A.

A. Ray, J. R. Rajian, Y. E. Lee, X. Wang, and R. Kopelman, J. Biomed. Opt. 17, 057004 (2012).
[CrossRef]

Santeford, A. C.

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

Shelton, R. L.

R. L. Shelton and B. E. Applegate, Biomed. Opt. Express 1, 676 (2010).
[CrossRef]

R. L. Shelton, S. P. Mattison, and B. E. Applegate, “Volumetric imaging of erythrocytes using label-free multiphoton photoacoustic microscopy,” J. Biophotonics, doi: 10.1002/jbio.201300059.
[CrossRef]

Shi, W.

Shi, Y. F.

Shohet, R. V.

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

Shung, K. K.

Song, W.

W. Song, Q. Wei, S. Jiao, and H. F. Zhang, J. Visualized Exp. 71, e4390 (2013).

T. Liu, H. Li, W. Song, S. L. Jiao, and H. F. Zhang, Curr. Eye Res. 38, 1229 (2013).
[CrossRef]

Stoica, G.

Suffredini, G.

G. Suffredini, J. E. East, and L. M. Levy, “New applications of nanotechnology for neuroimaging,” Am. J. Neuroradiol. (to be published).

Taber, L. A.

Wachowicz, K.

Wang, L. D.

L. D. Wang, K. Maslov, and L. H. V. Wang, Proc. Natl. Acad. Sci. USA 110, 5759 (2013).
[CrossRef]

Wang, L. H. V.

Wang, L. V.

L. V. Wang and S. Hu, Science 335, 1458 (2012).
[CrossRef]

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

S. Hu, K. Maslov, and L. V. Wang, Opt. Lett. 36, 269 (2011).

J. J. Yao, K. I. Maslov, Y. Zhang, Y. N. Xia, and L. V. Wang, J. Biomed. Opt. 16, 076003 (2011).
[CrossRef]

S. Hu and L. V. Wang, Front. Neuroenerg. 2, 10 (2010).

S. Hu, B. Rao, K. Maslov, and L. V. Wang, Opt. Lett. 35, 1 (2010).
[CrossRef]

K. Maslov, H. F. Zhang, S. Hu, and L. V. Wang, Opt. Lett. 33, 929 (2008).
[CrossRef]

Wang, X.

A. Ray, J. R. Rajian, Y. E. Lee, X. Wang, and R. Kopelman, J. Biomed. Opt. 17, 057004 (2012).
[CrossRef]

Wei, Q.

W. Song, Q. Wei, S. Jiao, and H. F. Zhang, J. Visualized Exp. 71, e4390 (2013).

Xia, Y. N.

J. J. Yao, K. I. Maslov, Y. Zhang, Y. N. Xia, and L. V. Wang, J. Biomed. Opt. 16, 076003 (2011).
[CrossRef]

Yao, J. J.

J. J. Yao, K. I. Maslov, Y. Zhang, Y. N. Xia, and L. V. Wang, J. Biomed. Opt. 16, 076003 (2011).
[CrossRef]

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

J. J. Yao, K. I. Maslov, Y. F. Shi, L. A. Taber, and L. H. V. Wang, Opt. Lett. 35, 1419 (2010).
[CrossRef]

Zemp, R. J.

Zhang, H. F.

Zhang, Y.

J. J. Yao, K. I. Maslov, Y. Zhang, Y. N. Xia, and L. V. Wang, J. Biomed. Opt. 16, 076003 (2011).
[CrossRef]

Zhou, Q. F.

Biomed. Opt. Express

Blood

S. S. Oladipupo, S. Hu, A. C. Santeford, J. J. Yao, J. R. Kovalski, R. V. Shohet, K. Maslov, L. V. Wang, and J. M. Arbeit, Blood 117, 4142 (2011).
[CrossRef]

Curr. Eye Res.

T. Liu, H. Li, W. Song, S. L. Jiao, and H. F. Zhang, Curr. Eye Res. 38, 1229 (2013).
[CrossRef]

Front. Neuroenerg.

S. Hu and L. V. Wang, Front. Neuroenerg. 2, 10 (2010).

J. Biomed. Opt.

J. J. Yao, K. I. Maslov, Y. Zhang, Y. N. Xia, and L. V. Wang, J. Biomed. Opt. 16, 076003 (2011).
[CrossRef]

Y. Jiang, A. Forbrich, T. Harrison, and R. J. Zemp, J. Biomed. Opt. 17, 036012 (2012).
[CrossRef]

A. Ray, J. R. Rajian, Y. E. Lee, X. Wang, and R. Kopelman, J. Biomed. Opt. 17, 057004 (2012).
[CrossRef]

J. Visualized Exp.

W. Song, Q. Wei, S. Jiao, and H. F. Zhang, J. Visualized Exp. 71, e4390 (2013).

Opt. Express

Opt. Lett.

Proc. Natl. Acad. Sci. USA

L. D. Wang, K. Maslov, and L. H. V. Wang, Proc. Natl. Acad. Sci. USA 110, 5759 (2013).
[CrossRef]

Science

L. V. Wang and S. Hu, Science 335, 1458 (2012).
[CrossRef]

Other

R. L. Shelton, S. P. Mattison, and B. E. Applegate, “Volumetric imaging of erythrocytes using label-free multiphoton photoacoustic microscopy,” J. Biophotonics, doi: 10.1002/jbio.201300059.
[CrossRef]

G. Suffredini, J. E. East, and L. M. Levy, “New applications of nanotechnology for neuroimaging,” Am. J. Neuroradiol. (to be published).

Supplementary Material (4)

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Figures (6)

Fig. 1.
Fig. 1.

Schematic of fully motorized OR-PAM.

Fig. 2.
Fig. 2.

Quantification of the lateral resolution of the FM-OR-PAM system. (a) Maximum-amplitude-projection (MAP) image of a sharp-edged metal square acquired with FM-OR-PAM. (b) Edge spread function (ESF) extracted from (a) and line spread function (LSF) obtained by taking the derivative of the ESF. The ESF data were averaged along y.

Fig. 3.
Fig. 3.

In vivo MAP images of (a) total hemoglobin, (b) sO2 (measured at 532 and 559 nm) in a mouse ear acquired with FM-OR-PAM in mode 2, and (c) 3D vasculature of the mouse ear (Media 1). (d) Blood flow distribution across the vessel pair marked with an arrow in (b) imaged in mode 1.

Fig. 4.
Fig. 4.

Comparison of 2D mechanical scanning at one depth (mode 2) and 3D mechanical contour scanning (mode 3) of a mouse ear in vivo. The white circles outline the tumor region. (a) Photo of a mouse ear with a growing tumor. In vivo MAP images acquired with FM-OR-PAM in (b) mode 2 and (c) mode 3; (d) 3D image of the tumorous ear vasculature (Media 2).

Fig. 5.
Fig. 5.

In vivo real-time imaging of single red blood cells (RBCs) in a mouse ear. (a) MAP image acquired with FM-OR-PAM in mode 2. (b) Tracking single RBCs in a selected capillary in vivo in mode 4 (2D optical scanning) (Media 3).

Fig. 6.
Fig. 6.

Mouse ear imaging in vivo with FM-OR-PAM in mode 5 (synchronized 1D optical and 2D mechanical hybrid scanning). (a) Mechanism of the mode 5 scanning. (b) MAP image of the mouse ear vasculature acquired in mode 5 (Media 4).

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