Abstract

A technique on high frame rate(28fps), high frequency co-registered ultrasound and photoacoustic imaging for visualizing zebrafish heart blood flow was demonstrated. This approach was achieved with a 40MHz light weight(0.38g) ring-type transducer, serving as the ultrasound transmitter and receiver, to allow an optic fiber, coupled with a 532nm laser, to be inserted into the hole. From the wire target study, axial resolutions of 38µm and 42µm were obtained for ultrasound and photoacoustic imaging, respectively. Carbon nanotubes were utilized as contrast agents to increase the flow signal level by 20dB in phantom studies, and zebrafish heart blood flow was successfully observed.

© 2013 OSA

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References

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  1. K. D. Poss, L. G. Wilson, and M. T. Keating, “Heart regeneration in Zebrafish,” Science298(5601), 2188–2190 (2002).
    [CrossRef] [PubMed]
  2. J. R. Hove, “In vivo biofluid dynamic imaging in the developing Zebrafish,” Birth Defects Res. C Embryo Today72(3Part C), 277–289 (2004).
    [CrossRef] [PubMed]
  3. J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
    [CrossRef] [PubMed]
  4. L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
    [CrossRef] [PubMed]
  5. J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
    [CrossRef] [PubMed]
  6. T. Harrison, J. C. Ranasinghesagara, H. Lu, K. Mathewson, A. Walsh, and R. J. Zemp, “Combined photoacoustic and ultrasound biomicroscopy,” Opt. Express17(24), 22041–22046 (2009).
    [CrossRef] [PubMed]
  7. Y. Zhang, H. Hong, and W. Cai, “Photoacoustic imaging,” Cold Spring Harb Protoc2011(9), 1015–1025 (2011).
    [CrossRef] [PubMed]
  8. J. Park, C. Hu, and K. K. Shung, “Stand-alone front-end system for high- frequency, high-frame-rate coded excitation ultrasonic imaging,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control58(12), 2620–2630 (2011).
    [CrossRef] [PubMed]
  9. J. M. Cannata, T. A. Ritter, W. H. Chen, R. H. Silverman, and K. K. Shung, “Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control50(11), 1548–1557 (2003).
    [CrossRef] [PubMed]
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    [PubMed]

2013

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

2012

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

2011

Y. Zhang, H. Hong, and W. Cai, “Photoacoustic imaging,” Cold Spring Harb Protoc2011(9), 1015–1025 (2011).
[CrossRef] [PubMed]

J. Park, C. Hu, and K. K. Shung, “Stand-alone front-end system for high- frequency, high-frame-rate coded excitation ultrasonic imaging,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control58(12), 2620–2630 (2011).
[CrossRef] [PubMed]

2009

2008

L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
[CrossRef] [PubMed]

2004

J. R. Hove, “In vivo biofluid dynamic imaging in the developing Zebrafish,” Birth Defects Res. C Embryo Today72(3Part C), 277–289 (2004).
[CrossRef] [PubMed]

2003

J. M. Cannata, T. A. Ritter, W. H. Chen, R. H. Silverman, and K. K. Shung, “Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control50(11), 1548–1557 (2003).
[CrossRef] [PubMed]

2002

K. D. Poss, L. G. Wilson, and M. T. Keating, “Heart regeneration in Zebrafish,” Science298(5601), 2188–2190 (2002).
[CrossRef] [PubMed]

Cai, W.

Y. Zhang, H. Hong, and W. Cai, “Photoacoustic imaging,” Cold Spring Harb Protoc2011(9), 1015–1025 (2011).
[CrossRef] [PubMed]

Cai, X.

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

Cannata, J. M.

J. M. Cannata, T. A. Ritter, W. H. Chen, R. H. Silverman, and K. K. Shung, “Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control50(11), 1548–1557 (2003).
[CrossRef] [PubMed]

Chen, R.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

Chen, W. H.

J. M. Cannata, T. A. Ritter, W. H. Chen, R. H. Silverman, and K. K. Shung, “Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control50(11), 1548–1557 (2003).
[CrossRef] [PubMed]

Cummins, T. M.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

Favazza, C.

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

Feng, C.

L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
[CrossRef] [PubMed]

Harrison, T.

Hong, H.

Y. Zhang, H. Hong, and W. Cai, “Photoacoustic imaging,” Cold Spring Harb Protoc2011(9), 1015–1025 (2011).
[CrossRef] [PubMed]

Hove, J. R.

J. R. Hove, “In vivo biofluid dynamic imaging in the developing Zebrafish,” Birth Defects Res. C Embryo Today72(3Part C), 277–289 (2004).
[CrossRef] [PubMed]

Hu, C.

J. Park, C. Hu, and K. K. Shung, “Stand-alone front-end system for high- frequency, high-frame-rate coded excitation ultrasonic imaging,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control58(12), 2620–2630 (2011).
[CrossRef] [PubMed]

Huang, Y.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

Johnson, J. A.

L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
[CrossRef] [PubMed]

Keating, M. T.

K. D. Poss, L. G. Wilson, and M. T. Keating, “Heart regeneration in Zebrafish,” Science298(5601), 2188–2190 (2002).
[CrossRef] [PubMed]

Lee, J.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

Li, P.

E. K. Pugach, P. Li, R. White, and L. Zon, “Retro-orbital injection in adult zebrafish,” J. Vis. Exp.34(34), 1645 (2009).
[PubMed]

Lien, C. L.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

Lu, H.

Maslov, K.

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

Mathewson, K.

Park, J.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

J. Park, C. Hu, and K. K. Shung, “Stand-alone front-end system for high- frequency, high-frame-rate coded excitation ultrasonic imaging,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control58(12), 2620–2630 (2011).
[CrossRef] [PubMed]

Poss, K. D.

K. D. Poss, L. G. Wilson, and M. T. Keating, “Heart regeneration in Zebrafish,” Science298(5601), 2188–2190 (2002).
[CrossRef] [PubMed]

Pugach, E. K.

E. K. Pugach, P. Li, R. White, and L. Zon, “Retro-orbital injection in adult zebrafish,” J. Vis. Exp.34(34), 1645 (2009).
[PubMed]

Ranasinghesagara, J. C.

Richard, W. D.

L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
[CrossRef] [PubMed]

Ritter, T. A.

J. M. Cannata, T. A. Ritter, W. H. Chen, R. H. Silverman, and K. K. Shung, “Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control50(11), 1548–1557 (2003).
[CrossRef] [PubMed]

Shung, K. K.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

J. Park, C. Hu, and K. K. Shung, “Stand-alone front-end system for high- frequency, high-frame-rate coded excitation ultrasonic imaging,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control58(12), 2620–2630 (2011).
[CrossRef] [PubMed]

L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
[CrossRef] [PubMed]

J. M. Cannata, T. A. Ritter, W. H. Chen, R. H. Silverman, and K. K. Shung, “Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control50(11), 1548–1557 (2003).
[CrossRef] [PubMed]

Silverman, R. H.

J. M. Cannata, T. A. Ritter, W. H. Chen, R. H. Silverman, and K. K. Shung, “Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control50(11), 1548–1557 (2003).
[CrossRef] [PubMed]

Sun, L.

L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
[CrossRef] [PubMed]

Walsh, A.

Wang, L. V.

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

White, R.

E. K. Pugach, P. Li, R. White, and L. Zon, “Retro-orbital injection in adult zebrafish,” J. Vis. Exp.34(34), 1645 (2009).
[PubMed]

Wilson, L. G.

K. D. Poss, L. G. Wilson, and M. T. Keating, “Heart regeneration in Zebrafish,” Science298(5601), 2188–2190 (2002).
[CrossRef] [PubMed]

Xu, X.

L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
[CrossRef] [PubMed]

Yang, J. M.

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

Yao, J.

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

Zemp, R. J.

Zhang, Y.

Y. Zhang, H. Hong, and W. Cai, “Photoacoustic imaging,” Cold Spring Harb Protoc2011(9), 1015–1025 (2011).
[CrossRef] [PubMed]

Zhou, Q.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

Zon, L.

E. K. Pugach, P. Li, R. White, and L. Zon, “Retro-orbital injection in adult zebrafish,” J. Vis. Exp.34(34), 1645 (2009).
[PubMed]

Ann. Biomed. Eng.

J. Park, Y. Huang, R. Chen, J. Lee, T. M. Cummins, Q. Zhou, C. L. Lien, and K. K. Shung, “Pulse Inversion Chirp Coded Tissue Harmonic Imaging (PI-CTHI) of zebrafish heart using high frame rate ultrasound biomicroscopy,” Ann. Biomed. Eng.41(1), 41–52 (2013).
[CrossRef] [PubMed]

Birth Defects Res. C Embryo Today

J. R. Hove, “In vivo biofluid dynamic imaging in the developing Zebrafish,” Birth Defects Res. C Embryo Today72(3Part C), 277–289 (2004).
[CrossRef] [PubMed]

Cold Spring Harb Protoc

Y. Zhang, H. Hong, and W. Cai, “Photoacoustic imaging,” Cold Spring Harb Protoc2011(9), 1015–1025 (2011).
[CrossRef] [PubMed]

IEEE Trans. Biomed. Eng.

L. Sun, X. Xu, W. D. Richard, C. Feng, J. A. Johnson, and K. K. Shung, “A high-frame rate duplex ultrasound biomicroscopy for small animal imaging in vivo,” IEEE Trans. Biomed. Eng.55(8), 2039–2049 (2008).
[CrossRef] [PubMed]

IEEE Trans. Ultrason. Ferroelectr. Freq. Control

J. Park, C. Hu, and K. K. Shung, “Stand-alone front-end system for high- frequency, high-frame-rate coded excitation ultrasonic imaging,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control58(12), 2620–2630 (2011).
[CrossRef] [PubMed]

J. M. Cannata, T. A. Ritter, W. H. Chen, R. H. Silverman, and K. K. Shung, “Design of efficient, broadband single-element (20-80 MHz) ultrasonic transducers for medical imaging applications,” IEEE Trans. Ultrason. Ferroelectr. Freq. Control50(11), 1548–1557 (2003).
[CrossRef] [PubMed]

J. Vis. Exp.

E. K. Pugach, P. Li, R. White, and L. Zon, “Retro-orbital injection in adult zebrafish,” J. Vis. Exp.34(34), 1645 (2009).
[PubMed]

Nat. Med.

J. M. Yang, C. Favazza, R. Chen, J. Yao, X. Cai, K. Maslov, Q. Zhou, K. K. Shung, and L. V. Wang, “Simultaneous functional photoacoustic and ultrasonic endoscopy of internal organs in vivo,” Nat. Med.18(8), 1297–1302 (2012).
[CrossRef] [PubMed]

Opt. Express

Science

K. D. Poss, L. G. Wilson, and M. T. Keating, “Heart regeneration in Zebrafish,” Science298(5601), 2188–2190 (2002).
[CrossRef] [PubMed]

Supplementary Material (10)

» Media 1: JPG (82 KB)     
» Media 2: JPG (68 KB)     
» Media 3: JPG (96 KB)     
» Media 4: JPG (95 KB)     
» Media 5: JPG (37 KB)     
» Media 6: JPG (38 KB)     
» Media 7: AVI (3871 KB)     
» Media 8: AVI (3871 KB)     
» Media 9: AVI (3191 KB)     
» Media 10: AVI (3191 KB)     

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Figures (6)

Fig. 1
Fig. 1

Block diagram of custom designed high frame rate imaging system capable of co-registered high frequency ultrasound and photoacoustic imaging. The trigger signal generator takes position signal and distributes trigger signals for acquiring photoacoustic and ultrasound signals simultaneously.

Fig. 2
Fig. 2

Beam sequences for the co-registered imaging. From the top row, trigger signals for transmitting ultrasound and laser and receiving echo signals for analog to digital converter are shown. In the last row, the alternating photoacoustic (PX) and ultrasound transmits (UX) are depicted.

Fig. 3
Fig. 3

(a) Picture and (b) mechanical design of the implemented light weight photoacoustic imaging transducer. (c) Pulse echo test result shown in time (solid line) and frequency (dotted line) domains.

Fig. 4
Fig. 4

Wire phantom study results. Images acquired by (a) ultrasound, (b) photoacoustic method. (c) Axial brightness profiles and (d) lateral brightness profiles are provided. Note that red solid line and blue dashed line indicate ultrasound and photoacoustic profiles respectively.

Fig. 5
Fig. 5

Coregistered ultrasound images (gray scale) with photoacoustic images (red scales). Images show cross sectional view of a plastic tube filled with (a) pure de-ionized water or (b) de-ionized water with carbon nano particles. Note that The scale of colorbar is dB.

Fig. 6
Fig. 6

Zebrafish heart co-registered ultrasound images (gray scale) with photoacoustic images (red scale) (a) with and (b) without SWNT contrast agent and (c) shows the co-registered image with contrast agent and static tissue signal suppression. Dotted arrows indicate blood flow found in the provided movie (Media 1). Note that the scale of colorbar is dB.

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