Objectives: Marginal donor kidneys are increasingly used for transplantation to overcome organ shortage. This study aims to investigate the additional value of Power Doppler (PD) imaging in kidney quality assessment during normothermic machine perfusion (NMP). Methods: Porcine kidneys (n = 22) retrieved from a local slaughterhouse underwent 2 h of NMP. Based on creatinine clearance (CrCl) and oxygen consumption (VO₂) at 120 min, kidneys were classified into Group 1 (n = 7, CrCl > 1 mL/min/100 g and VO₂ > 2.6 mL/min/100 g) and Group 2 (n = 15, CrCl ≤ 1 mL/min/100 g and/or VO₂ ≤ 2.6 mL/min/100 g). PD imaging was performed at 30, 60, and 120 min, and PD metrics, including vascularization index (VI), flow index (FI), and vascularization flow index (VFI) were calculated. Renal blood flow (RBF), CrCl, and VO₂ were measured at the same time points during NMP. The metrics were compared utilizing correlation analysis. Results: FI and VFI moderately correlated with CrCl (r = 0.537, p < 0.0001; r = 0.536, p < 0.0001, respectively), while VI strongly correlated with VO₂ (r = 0.839, p < 0.0001). At 120 min, PD metrics demonstrated the highest diagnostic accuracy for distinguishing between the two groups, with an area under the curve (AUC) of 0.943 for VI, 0.924 for FI, and 0.943 for VFI. Cutoff values of 17% for VI, 50 a.u. for FI, and 9 a.u. for VFI provided 100% specificity and 73% sensitivity in identifying kidneys in Group 2, with an overall diagnostic accuracy of 82%. Baseline kidney biopsies showed moderate acute tubular necrosis in both groups, with no significant differences. Conclusions: PD metrics strongly correlate with renal viability and effectively differentiate kidneys with higher and lower functionality during NMP. PD imaging can be a valuable alternative to RBF during NMP for kidney quality assessment.

Photoacoustic imaging offers optical contrast images of human tissue at acoustic resolution, making it valuable for diverse clinical applications. However, quantifying tissue composition via optical contrast remains challenging due to the unknown light fluence within the tissue. Here, we propose a method that leverages known chromophores (e.g., arterial blood) to improve the accuracy of quantitative photoacoustic imaging. By using the optical properties of a known chromophore as a fluence marker and integrating it into the optical inversion process, we can estimate the unknown fluence within the tissue. Experimentally, we demonstrate that this approach successfully recovers both the spectral shape and magnitude of the optical absorption coefficient of an unknown chromophore. Additionally, we show that the fluence marker method enhances conventional optical inversion techniques, specifically (i) a straightforward iterative approach and (ii) a gradient-based method. Our results indicate an improvement in accuracy of up to 24.4% when comparing optical absorption recovery with and without the fluence marker. Finally, we present the method's performance and illustrate its applications in carotid plaque quantification.

Mass spectrometry imaging (MSI) is a powerful tool for detecting lipids in tissue sections, with matrix-assisted laser desorption/ionization (MALDI) and desorption electrospray ionization (DESI) as its key ionization techniques. In this study, we examine how MALDI compares with state-of-the-art DESI ionization in identifying lipids in heterogeneous samples, specifically atherosclerotic plaques. Carotid plaques (n = 4) from patients undergoing endarterectomy were snap-frozen, stored at −80°C, and then sectioned for MSI analysis and H&E staining. Measurements were conducted using a SYNAPT XS mass spectrometer in positive ion mode, employing MALDI with a 2,5-dihydroxybenzoic acid (DHB) matrix and DESI with a methanol: water (98:2) (v/v) solvent. Our comparison covered spectral profiles, sensitivity, and image quality generated by these two techniques. We found that both MALDI and DESI are highly suitable techniques for detecting a wide range of lipids in atherosclerotic plaque sections. DESI-MSI exhibited higher ion counts for most lipid classes than MALDI-MSI and provided sharper images. MALDI detected larger amounts of ceramide and hexosylceramide species, possibly due to its efficient generation of dehydrated ions. In contrast, DESI showed greater peak intensities of cholesteryl ester and triacylglyceride species than MALDI, consistent with reduced fragmentation. These findings establish the relative merits of DESI and MALDI and demonstrate their complementarity as techniques for lipid research in MSI.

Spectral photoacoustic imaging in combination with unmixing techniques may be applied to retrieve information about high-risk features present in atherosclerotic plaques, possibly providing prognostic insights into future stroke events. We present the photoacoustic spectral contrast found in 12 systematically scanned advanced atherosclerotic plaques in the near-infrared wavelength range (850–1250 nm). The main absorbers are lipid, water, and hemoglobin, with the highest photoacoustic intensities at the lipid's second overtone at 1190 and 1210 nm. Linear unmixing resulted in visualizing regions with high lipid and hemoglobin absorption, corresponding to the histological presence of lipid and intraplaque hemorrhage. A non-negative matrix factorization approach reveals differences in lipid spectral contrast, providing potential insights into the vulnerability of atherosclerotic plaque. These results provide a reference for future, more complex, in vivo photoacoustic imaging of carotid artery atherosclerosis, potentially contributing to assessing the risk of future events and treatment decision.