Abstract

Compared with visible light (380–700 nm), near-infrared light (700–1400 nm) undergoes weaker optical attenuation in biological tissue; thus, it can penetrate deeper. Herein, we demonstrate near-infrared optical-resolution photoacoustic microscopy (NIR-OR-PAM) with 1046 nm illumination. A penetration depth of 3.2 mm was achieved in chicken breast tissue ex vivo using optical fluence within the American National Standards Institute (ANSI) limit (100mJ/cm2). Beyond 0.6mm deep in chicken breast tissue, NIR-OR-PAM has shown finer resolution than the visible counterpart with 570 nm illumination. The deep imaging capability of NIR-OR-PAM was validated in both a mouse ear and a mouse brain. NIR-OR-PAM of possible lipid contrast was explored as well.

© 2014 Optical Society of America

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References

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T. Matthews, C. Zhang, D. Yao, K. Maslov, and L. V. Wang, J. Biomed. Opt. 19, 016004 (2014).
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2013 (3)

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J. Yao and L. V. Wang, Laser Photonics Rev. 7, 758 (2013).
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2012 (4)

T. J. Allen, A. Hall, A. P. Dhillon, J. S. Owen, and P. C. Beard, J. Biomed. Opt. 17, 061209 (2012).
[CrossRef]

L. V. Wang and S. Hu, Science 335, 1458 (2012).
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Y. Liu, C. Zhang, and L. V. Wang, J. Biomed. Opt. 17, 126014 (2012).
[CrossRef]

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

2011 (4)

2010 (2)

2008 (2)

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2006 (1)

H. F. Zhang, K. Maslov, G. Stoica, and L. V. Wang, Nat. Biotechnol. 24, 848 (2006).
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S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

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T. J. Allen, A. Hall, A. P. Dhillon, J. S. Owen, and P. C. Beard, J. Biomed. Opt. 17, 061209 (2012).
[CrossRef]

Arai, T.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Ashida, H.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

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T. J. Allen, A. Hall, A. P. Dhillon, J. S. Owen, and P. C. Beard, J. Biomed. Opt. 17, 061209 (2012).
[CrossRef]

Carson, P. L.

Chen, S.

Chen, Y.-S.

Cubeddu, R.

R. L. P. van Veen, H. J. C. M. Sterenborg, A. Pifferi, A. Torricelli, and R. Cubeddu, in Biomedical Topical Meeting, OSA Technical Digest (2004), paper SF4.

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T. J. Allen, A. Hall, A. P. Dhillon, J. S. Owen, and P. C. Beard, J. Biomed. Opt. 17, 061209 (2012).
[CrossRef]

Emelianov, S.

Forbrich, A. E.

Freudiger, C. W.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

Gao, L.

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

Guo, L. J.

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Hall, A.

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[CrossRef]

He, C.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

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Holtom, G. R.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

Homan, K.

Hu, S.

Huang, C. H.

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

Ishihara, M.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

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C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

Kawauchi, S.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Kikuchi, M.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Kim, S.

Kurita, A.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Li, C.

J. Yao, L. Wang, C. Li, C. Zhang, and L. V. Wang, Phys. Rev. Lett. 112, 014302 (2014).
[CrossRef]

Li, C. H.

Z. Xu, C. H. Li, and L. V. Wang, J. Biomed. Opt. 15, 036019 (2010).
[CrossRef]

Liang, J.

J. Liang, Y. Zhou, K. I. Maslov, and L. V. Wang, J. Biomed. Opt. 18, 096004 (2013).
[CrossRef]

Ling, T.

Liu, Y.

Y. Liu, C. Zhang, and L. V. Wang, J. Biomed. Opt. 17, 126014 (2012).
[CrossRef]

Lu, S.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

Mallidi, S.

Maslov, K.

T. Matthews, C. Zhang, D. Yao, K. Maslov, and L. V. Wang, J. Biomed. Opt. 19, 016004 (2014).
[CrossRef]

L. Wang, K. Maslov, J. Yao, B. Rao, and L. V. Wang, Opt. Lett. 36, 139 (2011).
[CrossRef]

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

H. F. Zhang, K. Maslov, G. Stoica, and L. V. Wang, Nat. Biotechnol. 24, 848 (2006).
[CrossRef]

Maslov, K. I.

J. Liang, Y. Zhou, K. I. Maslov, and L. V. Wang, J. Biomed. Opt. 18, 096004 (2013).
[CrossRef]

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

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

Matsui, T.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Matthews, T.

T. Matthews, C. Zhang, D. Yao, K. Maslov, and L. V. Wang, J. Biomed. Opt. 19, 016004 (2014).
[CrossRef]

Min, W.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

Obara, M.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Ogura, M.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Owen, J. S.

T. J. Allen, A. Hall, A. P. Dhillon, J. S. Owen, and P. C. Beard, J. Biomed. Opt. 17, 061209 (2012).
[CrossRef]

Paproski, R. J.

Pifferi, A.

R. L. P. van Veen, H. J. C. M. Sterenborg, A. Pifferi, A. Torricelli, and R. Cubeddu, in Biomedical Topical Meeting, OSA Technical Digest (2004), paper SF4.

Querry, M. R.

Rao, B.

Saar, B. G.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

Sato, S.

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Sterenborg, H. J. C. M.

R. L. P. van Veen, H. J. C. M. Sterenborg, A. Pifferi, A. Torricelli, and R. Cubeddu, in Biomedical Topical Meeting, OSA Technical Digest (2004), paper SF4.

Stoica, G.

H. F. Zhang, K. Maslov, G. Stoica, and L. V. Wang, Nat. Biotechnol. 24, 848 (2006).
[CrossRef]

Torricelli, A.

R. L. P. van Veen, H. J. C. M. Sterenborg, A. Pifferi, A. Torricelli, and R. Cubeddu, in Biomedical Topical Meeting, OSA Technical Digest (2004), paper SF4.

Tsai, J. C.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

van Veen, R. L. P.

R. L. P. van Veen, H. J. C. M. Sterenborg, A. Pifferi, A. Torricelli, and R. Cubeddu, in Biomedical Topical Meeting, OSA Technical Digest (2004), paper SF4.

Wachowicz, K.

Wang, B.

Wang, L.

J. Yao, L. Wang, C. Li, C. Zhang, and L. V. Wang, Phys. Rev. Lett. 112, 014302 (2014).
[CrossRef]

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

L. Wang, K. Maslov, J. Yao, B. Rao, and L. V. Wang, Opt. Lett. 36, 139 (2011).
[CrossRef]

Wang, L. V.

J. Yao, L. Wang, C. Li, C. Zhang, and L. V. Wang, Phys. Rev. Lett. 112, 014302 (2014).
[CrossRef]

T. Matthews, C. Zhang, D. Yao, K. Maslov, and L. V. Wang, J. Biomed. Opt. 19, 016004 (2014).
[CrossRef]

J. Liang, Y. Zhou, K. I. Maslov, and L. V. Wang, J. Biomed. Opt. 18, 096004 (2013).
[CrossRef]

C. Zhang, Y. Zhang, D. K. Yao, Y. Xia, and L. V. Wang, J. Biomed. Opt. 18, 020504 (2013).
[CrossRef]

J. Yao and L. V. Wang, Laser Photonics Rev. 7, 758 (2013).
[CrossRef]

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

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

Y. Liu, C. Zhang, and L. V. Wang, J. Biomed. Opt. 17, 126014 (2012).
[CrossRef]

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

L. Wang, K. Maslov, J. Yao, B. Rao, and L. V. Wang, Opt. Lett. 36, 139 (2011).
[CrossRef]

Z. Xu, C. H. Li, and L. V. Wang, J. Biomed. Opt. 15, 036019 (2010).
[CrossRef]

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

H. F. Zhang, K. Maslov, G. Stoica, and L. V. Wang, Nat. Biotechnol. 24, 848 (2006).
[CrossRef]

Wang, X.

Xia, Y.

C. Zhang, Y. Zhang, D. K. Yao, Y. Xia, and L. V. Wang, J. Biomed. Opt. 18, 020504 (2013).
[CrossRef]

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

Xie, X. S.

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

Xie, Z.

Xu, Z.

Z. Xu, C. H. Li, and L. V. Wang, J. Biomed. Opt. 15, 036019 (2010).
[CrossRef]

Yang, J. M.

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

Yao, D.

T. Matthews, C. Zhang, D. Yao, K. Maslov, and L. V. Wang, J. Biomed. Opt. 19, 016004 (2014).
[CrossRef]

Yao, D. K.

C. Zhang, Y. Zhang, D. K. Yao, Y. Xia, and L. V. Wang, J. Biomed. Opt. 18, 020504 (2013).
[CrossRef]

Yao, J.

J. Yao, L. Wang, C. Li, C. Zhang, and L. V. Wang, Phys. Rev. Lett. 112, 014302 (2014).
[CrossRef]

J. Yao and L. V. Wang, Laser Photonics Rev. 7, 758 (2013).
[CrossRef]

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

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

L. Wang, K. Maslov, J. Yao, B. Rao, and L. V. Wang, Opt. Lett. 36, 139 (2011).
[CrossRef]

Zemp, R. J.

Zhang, C.

J. Yao, L. Wang, C. Li, C. Zhang, and L. V. Wang, Phys. Rev. Lett. 112, 014302 (2014).
[CrossRef]

T. Matthews, C. Zhang, D. Yao, K. Maslov, and L. V. Wang, J. Biomed. Opt. 19, 016004 (2014).
[CrossRef]

C. Zhang, Y. Zhang, D. K. Yao, Y. Xia, and L. V. Wang, J. Biomed. Opt. 18, 020504 (2013).
[CrossRef]

Y. Liu, C. Zhang, and L. V. Wang, J. Biomed. Opt. 17, 126014 (2012).
[CrossRef]

Zhang, H. F.

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

H. F. Zhang, K. Maslov, G. Stoica, and L. V. Wang, Nat. Biotechnol. 24, 848 (2006).
[CrossRef]

Zhang, Y.

C. Zhang, Y. Zhang, D. K. Yao, Y. Xia, and L. V. Wang, J. Biomed. Opt. 18, 020504 (2013).
[CrossRef]

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

Zhou, Y.

J. Liang, Y. Zhou, K. I. Maslov, and L. V. Wang, J. Biomed. Opt. 18, 096004 (2013).
[CrossRef]

Zou, J.

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

Appl. Opt. (1)

Biomed. Opt. Express (1)

J. Biomed. Opt. (8)

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

T. Matthews, C. Zhang, D. Yao, K. Maslov, and L. V. Wang, J. Biomed. Opt. 19, 016004 (2014).
[CrossRef]

J. Yao, C. H. Huang, L. Wang, J. M. Yang, L. Gao, K. I. Maslov, J. Zou, and L. V. Wang, J. Biomed. Opt. 17, 0805051 (2012).

J. Liang, Y. Zhou, K. I. Maslov, and L. V. Wang, J. Biomed. Opt. 18, 096004 (2013).
[CrossRef]

Y. Liu, C. Zhang, and L. V. Wang, J. Biomed. Opt. 17, 126014 (2012).
[CrossRef]

C. Zhang, Y. Zhang, D. K. Yao, Y. Xia, and L. V. Wang, J. Biomed. Opt. 18, 020504 (2013).
[CrossRef]

Z. Xu, C. H. Li, and L. V. Wang, J. Biomed. Opt. 15, 036019 (2010).
[CrossRef]

T. J. Allen, A. Hall, A. P. Dhillon, J. S. Owen, and P. C. Beard, J. Biomed. Opt. 17, 061209 (2012).
[CrossRef]

Laser Photonics Rev. (1)

J. Yao and L. V. Wang, Laser Photonics Rev. 7, 758 (2013).
[CrossRef]

Lasers Surg. Med. (1)

S. Sato, M. Ogura, M. Ishihara, S. Kawauchi, T. Arai, T. Matsui, A. Kurita, M. Obara, M. Kikuchi, and H. Ashida, Lasers Surg. Med. 29, 464 (2001).
[CrossRef]

Nat. Biotechnol. (1)

H. F. Zhang, K. Maslov, G. Stoica, and L. V. Wang, Nat. Biotechnol. 24, 848 (2006).
[CrossRef]

Opt. Express (1)

Opt. Lett. (3)

Phys. Rev. Lett. (1)

J. Yao, L. Wang, C. Li, C. Zhang, and L. V. Wang, Phys. Rev. Lett. 112, 014302 (2014).
[CrossRef]

Science (2)

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

C. W. Freudiger, W. Min, B. G. Saar, S. Lu, G. R. Holtom, C. He, J. C. Tsai, J. X. Kang, and X. S. Xie, Science 322, 1857 (2008).
[CrossRef]

Other (2)

R. L. P. van Veen, H. J. C. M. Sterenborg, A. Pifferi, A. Torricelli, and R. Cubeddu, in Biomedical Topical Meeting, OSA Technical Digest (2004), paper SF4.

“American national standard for the safe use of lasers,” (2007).

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Figures (7)

Fig. 1.
Fig. 1.

Absorption spectra of four tissue components and the schematic of near-infrared optical-resolution photoacoustic microscopy (NIR-OR-PAM). (a) Absorption spectra of oxyhemoglobin (HbO2) (150g/L or 2.3 mM in blood), deoxyhemoglobin (HbR) (150g/L or 2.3 mM in blood), water (80% by volume in tissue), and lipid (20% by volume in tissue) from 250 to 1150 nm. (b) Schematic of the NIR-OR-PAM system. ConL, condenser lens; CorL, correction lens; DC, dye cell; DM, dichroic mirror; FM, flip mirror; M1, M2, and M3, mirrors; ND, neutral density filter; OL, objective lens; PH, pinhole; PL, pump laser; RAP, right-angle prism; RhP, rhomboid prism; SHG, second harmonic generator; SOL, silicone oil layer; UT, ultrasonic transducer; WT, water tank.

Fig. 2.
Fig. 2.

Penetration depths of the two optical-resolution photoacoustic microscopy (OR-PAM) systems. Images of a black human hair inserted obliquely into a piece of fresh chicken breast tissue. (a) Visible optical-resolution photoacoustic microscopy (VIS-OR-PAM). (b) Near-infrared optical-resolution photoacoustic microscopy (NIR-OR-PAM). Hair was imaged with a SNR of 6dB up to 3.2 mm deep in the tissue by NIR-OR-PAM and 2.3 mm deep in the tissue by VIS-OR-PAM.

Fig. 3.
Fig. 3.

Lateral resolutions of the two optical-resolution photoacoustic microscopy (OR-PAM) systems. Line spread functions (black dotted lines) extracted from the fitted edge spread functions (solid red lines) acquired in water. (a) Visible optical-resolution photoacoustic microscopy (VIS-OR-PAM). (b) Near-infrared optical-resolution photoacoustic microscopy (NIR-OR-PAM). Here, the lateral resolution was quantified as the full width at half maximum (FWHM) of the line spread functions. Lateral resolution of NIR-OR-PAM in water is 6.2 μm, while lateral resolution of VIS-OR-PAM is 2.9 μm. (c) Lateral resolution of NIR-OR-PAM and VIS-OR-PAM as a function of imaging depth in scattering media. Here, the lateral resolution was quantified as the FWHM of the numerical first derivative of the measured edge spread function.

Fig. 4.
Fig. 4.

Comparison of images of a mouse ear acquired using visible (VIS) and near-infrared (NIR) light in vivo. (a) Maximum amplitude projection (MAP) image of a mouse ear acquired by visible optical-resolution photoacoustic microscopy (VIS-OR-PAM) in vivo within depths up to 300 μm. Two close-ups show the photoacoustic (PA) signals from blood vessels, as indicated by the dashed boxes. (b) MAP image of the same mouse ear acquired by near-infrared optical-resolution photoacoustic microscopy (NIR-OR-PAM) in vivo within depths up to 300 μm. (c) and (d) B-scan images along the dashed lines in (a) and (b), respectively, on a logarithmic scale after acoustic attenuation compensation with an acoustic attenuation coefficient of 1dB/MHz/cm.

Fig. 5.
Fig. 5.

Comparison of in vivo imaging depths in a mouse brain. (a) xz projection of a visible optical-resolution photoacoustic microscopy (VIS-OR-PAM) image of the mouse brain acquired in vivo. (b) xy projections of the mouse brain at different depths. No structure can be detected at depths beyond 500 μm. (c) xz projection of a near-infrared optical-resolution photoacoustic microscopy (NIR-OR-PAM) image of the same mouse brain acquired in vivo. (d) xy projections of the mouse brain at different depths. Structures can be detected at depths up to 750 μm. Dashed lines in the xz projections mark the approximate location of the skull.

Fig. 6.
Fig. 6.

Comparison of in vivo lateral resolutions in mouse brain. (a) In vivo mouse brain maximum amplitude projection (MAP) image acquired by visible optical-resolution photoacoustic microscopy (VIS-OR-PAM). (b) In vivo mouse brain maximum amplitude projection (MAP) image acquired by near-infrared optical-resolution photoacoustic microscopy (NIR-OR-PAM). (c) Close-ups of the dashed boxes in (a) and (b) show that more blood vessels can be imaged by NIR-OR-PAM. (d) Normalized photoacoustic (PA) amplitudes across two selected blood vessels at 225 μm (solid lines) and 330 μm (dashed lines) depths in (c), respectively. Both vessels have smaller apparent diameters in the NIR-OR-PAM image than in the VIS-OR-PAM image because they are less blurred.

Fig. 7.
Fig. 7.

Comparison of penetrations in blood. (a) Top: visible optical-resolution photoacoustic microscopy (VIS-OR-PAM) maximum amplitude projection (MAP) image of a mouse brain acquired in vivo within depths up to 300 μm in which blood vessels were covered by a bleeding layer. Bottom: VIS-OR-PAM MAP image of the same area in the mouse brain acquired in vivo within depths of 100–300 μm after digitally peeling off the bleeding layer. Underlying blood vessels can barely be resolved. (b) Top: near-infrared optical-resolution photoacoustic microscopy (NIR-OR-PAM) MAP image of the same region as (a) within depths up to 300 μm. Signal strength from the bleeding layer is weaker than that from the VIS-OR-PAM counterpart. Bottom: NIR-OR-PAM MAP image of the same area in the mouse brain acquired in vivo within depths of 100–300 μm after digitally peeling off the bleeding layer. Underlying blood vessels can be clearly resolved.

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