Abstract
Three-dimensional (3D) optical imaging of whole biological organs with microscopic resolution has remained a challenge. Most versions of such imaging techniques require special preparation of the tissue specimen. Here we demonstrate microtomy-assisted photoacoustic microscopy (mPAM) of mouse brains and other organs, which automatically acquires serial distortion-free and registration-free images with endogenous absorption contrasts. Without tissue staining or clearing, mPAM generates micrometer-resolution 3D images of paraffin- or agarose-embedded whole organs with high fidelity, achieved by label-free simultaneous sensing of DNA/RNA, hemoglobins, and lipids. mPAM provides histology-like imaging of cell nuclei, blood vessels, axons, and other anatomical structures, enabling the application of histopathological interpretation at the organelle level to analyze a whole organ. Its deep tissue imaging capability leads to less sectioning, resulting in negligible sectioning artifact. mPAM offers a new way to better understand complex biological organs.
Highlights
Three-dimensional (3D) optical imaging of whole biological organs with microscopic resolution has remained a challenge
Among all 3D imaging techniques, histology is an attractive way to analyze specimens because histopathological interpretation can be readily applied from organelle to organ levels
In microtomy-assisted photoacoustic microscopy (mPAM) (Fig. 1a), an organ or a tissue block, either formalin-fixed or fresh, and paraffin or agarose embedded, is mounted on an organ holder immersed in water
Summary
Three-dimensional (3D) optical imaging of whole biological organs with microscopic resolution has remained a challenge. We propose a new label-free 3D imaging technique, named microtomy-assisted photoacoustic microscopy (mPAM), for potentially imaging all biomolecules with 100% endogenous natural staining in whole organs with less sectioning and high fidelity. Label-free PAM has been demonstrated in such broad biomedical applications as imaging DNA/RNA15, cytochromes[16], hemoglobins[17], melanin[18], and lipids[19] at an optical wavelength ranging from ultra-violet (UV) to near-infrared. This list of imaging targets is expanding with the ongoing exploration of endogenous absorption, and label-free imaging of all biomolecules remains possible. We demonstrate the first label-free mPAM, using UV light for histology-like a
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