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Published February 2012 | Published
Journal Article Open

Reflection-mode submicron-resolution in vivo photoacoustic microscopy


Submicron-resolution photoacoustic microscopy (PAM) currently exists only in transmission mode, due to the technical difficulties of combining high numerical-aperture (NA) optical illumination with high NA acoustic detection. The lateral resolution of reflection-mode PAM has not reached <2  μm in the visible light range. Here we develop the first reflection-mode submicron-resolution PAM system with a new compact design. By using a parabolic mirror to focus and reflect the photoacoustic waves, sufficient signals were collected for good sensitivity without distorting the optical focusing. By imaging nanospheres and a resolution test chart, the lateral resolution was measured to be ∼0.5  μm with an optical wavelength of 532 nm, an optical NA of 0.63. The axial resolution was measured at 15 μm. Here the axial resolution was measured by a different experiment with the lateral resolution measurement. But we didn't describe the details of axial resolution measurement due to space limit. The maximum penetration was measured at ∼0.42  mm in optical-scattering soft tissue. As a comparison, both the submicron-resolution PAM and a 2.4 μm-resolution PAM were used to image a mouse ear in vivo with the same optical wavelength and similar pulse energy. Capillaries were resolved better by the submicron-resolution PAM. Therefore, the submicron-resolution PAM is suitable for in vivo high-resolution imaging, or even subcellular imaging, of optical absorption.

Additional Information

© 2012 SPIE. Paper 11593L received Oct. 11, 2011; revised manuscript received Nov. 19, 2011; accepted for publication Dec. 29, 2011; published online Feb. 23, 2012. We thank Lidai Wang, Junjie Yao, Amy Winkler, and Bin Rao for experimental assistance and helpful discussions. This work was sponsored in part by National Institutes of Health grants R01 EB000712, R01 EB008085, R01 CA134539, U54 CA136398, R01 CA157277, and 5P60 DK02057933. Lihong Wang has a financial interest in Microphotoacoustics, Inc. and Endra, Inc., which, however, did not support this work.

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