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Published July 28, 2014 | Accepted Version + Supplemental Material
Journal Article Open

Optical-Resolution Photoacoustic Microscopy for Volumetric and Spectral Analysis of Histological and Immunochemical Samples


Optical-resolution photoacoustic microscopy (OR-PAM) is an imaging modality with superb penetration depth and excellent absorption contrast. Here we demonstrate, for the first time, that this technique can advance quantitative analysis of conventional chromogenic histochemistry. Because OR-PAM can quantify the absorption contrast at different wavelengths, it is feasible to spectrally resolve the specific biomolecules involved in a staining color. Furthermore, the tomographic capability of OR-PAM allows for noninvasive volumetric imaging of a thick sample without microtoming it. By immunostaining the sample with different chromogenic agents, we further demonstrated the ability of OR-PAM to resolve different types of cells in a coculture sample with imaging depths up to 1 mm. Taken together, the integration of OR-PAM with (immuno)histochemistry offers a simple and versatile technique with broad applications in cell biology, pathology, tissue engineering, and related biomedical studies.

Additional Information

© 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. Received: March 28, 2014; Published online: June 24, 2014. This work was supported by NIH grants DP1 OD000798 (NIH Director's Pioneer Award) to Y.X., and DP1 EB016986 (NIH Director's Pioneer Award), R01 CA186567 (NIH Director's Transformative Research Award), and R01 CA159959 to L.V.W. L.V.W. has a financial interest in Endra, Inc., and Microphotoacoustics, Inc., which did not support this work. Part of the research was performed at the Alafi Neuroimaging Laboratory, the Hope Center for Neurological Disorders in Washington University School of Medicine, which is supported by the NIH Neuroscience Blueprint Center Core Grant P30 NS057105.

Attached Files

Accepted Version - nihms588353.pdf

Supplemental Material - anie_201403812_sm_miscellaneous_information.pdf

Supplemental Material - anie_201403812_sm_movie_s1_scaffold_3d.wmv

Supplemental Material - anie_201403812_sm_movie_s2_co-culture_3d.wmv


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August 20, 2023
October 20, 2023