Conventional optoacoustic microscopy operates in two distinct modes of optical resolution, for visualization of superficial tissue layers, or acoustic resolution, intended for deep imaging in scattering tissues. Here we introduce a new microscope design with hybrid optical and acoustic resolution, which provides a smooth transition from optical resolution in superficial microscopic imaging to ultrasonic resolution when imaging at greater depths within intensely scattering tissue layers. Experimental validation of the new hybrid optoacoustic microscopy method was performed in phantoms and by means of transcranial mouse brain imaging in vivo.
Synthetic aperture focusing technique (SAFT) is effective in restoring lateral resolution of ultrasonic images for scans with focusing-related distortions. Although successfully applied in pulse-echo ultrasonics, the physical nature of an optoacoustic modality requires a modified algorithm to return accurate results. The SIR-SAFT method reported here uses the spatial impulse response (SIR) of the transducer to weight the contributions to the SAFT and is tailored to provide significant resolution and signal gains for out-of-focus sources in scanning optoacoustic microscopy systems. Furthermore, the SIR-SAFT is implemented in full three dimensions, applicable to signals both far of and at the focus of the ultrasonic detector. The method has been further shown to outperform conventional SAFT algorithms for both simulated and experimental optoacoustic data.
Despite the great promise behind the recent introduction of optoacoustic technology into the arsenal of small‐animal neuroimaging methods, a variety of acoustic and light‐related effects introduced by adult murine skull severely compromise the performance of optoacoustics in transcranial imaging. As a result, high‐resolution noninvasive optoacoustic microscopy studies are still limited to a thin layer of pial microvasculature, which can be effectively resolved by tight focusing of the excitation light. We examined a range of distortions introduced by an adult murine skull in transcranial optoacoustic imaging under both acoustically‐ and optically‐determined resolution scenarios. It is shown that strong low‐pass filtering characteristics of the skull may significantly deteriorate the achievable spatial resolution in deep brain imaging where no light focusing is possible. While only brain vasculature with a diameter larger than 60 µm was effectively resolved via transcranial measurements with acoustic resolution, significant improvements are seen through cranial windows and thinned skull experiments.
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