Single photon emission computed tomography (SPECT) can enable the quantification of activity uptake in lesions and at-risk organs in α-particle-emitting radiopharmaceutical therapies (α-RPTs). However, this quantification is challenged by the extremely low detected photon counts, complicated isotope physics, and the image-degrading effects in α-RPT SPECT. Thus, strategies to optimize the SPECT system and protocol designs for the task of regional uptake quantification are much needed. Objectively performing this task-based optimization requires a reliable (accurate and precise) regional uptake quantification method. Conventional reconstruction-based quantification (RBQ) methods have been observed to be erroneous for α-RPT SPECT. Projection-domain quantification methods, which estimate regional uptake directly from SPECT projections, have demonstrated potential in providing reliable regional uptake estimates, but these methods assume constant uptake within the regions, an assumption that may not hold. To address these challenges, we propose Wiener INtegration Projection-Domain Quantification (WIN-PDQ), a Wiener-estimator-based projection-domain quantitative SPECT method. The method accounts for the heterogeneity within the regions of interest while estimating mean uptake. An early-stage evaluation of the method was conducted using 3D Monte Carlo-simulated SPECT of anthropomorphic phantoms with 223Ra uptake and lumpy-model-based intra-regional uptake heterogeneity. In this evaluation with phantoms of varying mean regional uptake and intra-regional uptake heterogeneity, the WIN-PDQ method yielded ensemble unbiased estimates and significantly outperformed both reconstruction-based and previously proposed projection-domain quantification methods in terms of normalized root ensemble mean squared error. In conclusion, based on these preliminary findings, the proposed WIN-PDQ method is showing potential for estimating mean regional uptake in α-RPTs and towards enabling the objective task-based optimization of SPECT system and protocol designs.
Imaging Mass Cytometry (IMC) is an emerging multiplexed imaging pathology technology which can detect the spatial distribution of up to 40 markers at cellular resolution. However, the high content and complexity of this spatial information is underutilized in downstream analysis. To overcome this limitation, we develop an interpretable graph convolutional network (GCN) trained by IMC data with 30 cell markers from 238 patient breast cancer samples, with both marker expressions and cell locations. The network enables triple negative breast cancer (TNBC) classification from other clinical types, including HR+HER2+, HR+HER- and HR-HER+. More importantly, with an embedded self-attention pooling module, cell communities with high diagnostic values can be detected based on the attention scores. The proposed GCN framework is benchmarked with a fully connected artifical neural network (ANN) without spatial information. With a stratified 8-fold cross validation, GCN performs slightly better than ANN for tissue-level classification (class balanced accuracy: 0.8283±0.0964 to 0.8123±0.0989; area under the curve: 0.8548±0.1252 to 0.8298±0.1407). Nevertheless, GCN remarkably outperforms ANN on potentially interested cell community detection, especially in TNBC tissues, regarding the spearman correlation coefficient (SCC) between attention scores and marker expressions. The average SCC differences between GCN and ANN range from 0.0532 to 0.1876 for Cytokeratin 5, 7, 14, 8/18, 19, and pan Cytokeration. With comparisons of selected markers on tissues with different clinical types and attention scores, the cell marker expressions correlate with their clinical types and diagnostic values, which further validate the proposed framework. Overall, our GCN enables interpretable triple negative breast cancer detection and has the potential to be widely implemented in other diseases and highly multiplexed imaging techniques for enhanced microenvironment analysis.
Voltage-sensitive dyes (VSDs) are designed to monitor membrane potential by detecting fluorescence changes in response to neuronal or muscle electrical activity. However, fluorescence imaging is limited by depth of penetration and high scattering losses, which leads to low sensitivity in vivo systems for external detection. By contrast, photoacoustic (PA) imaging, an emerging modality, is capable of deep tissue, noninvasive imaging by combining near-infrared light excitation and ultrasound detection. Here, we show that voltage-dependent quenching of dye fluorescence leads to a reciprocal enhancement of PA intensity. We synthesized a near-infrared photoacoustic VSD (PA-VSD), whose PA intensity change is sensitive to membrane potential. In the polarized state, this cyanine-based probe enhances PA intensity while decreasing fluorescence output in a lipid vesicle membrane model. A theoretical model accounts for how the experimental PA intensity change depends on fluorescence and absorbance properties of the dye. These results not only demonstrate PA voltage sensing but also emphasize the interplay of both fluorescence and absorbance properties in the design of optimized PA probes. Together, our results demonstrate PA sensing as a potential new modality for recording and external imaging of electrophysiological and neurochemical events in the brain.
Monitoring of the membrane potential is possible using voltage sensitive dyes (VSD), where fluorescence intensity changes in response to neuronal electrical activity. However, fluorescence imaging is limited by depth of penetration and high scattering losses, which leads to low sensitivity in vivo systems for external detection. In contrast, photoacoustic (PA) imaging, an emerging modality, is capable of deep tissue, noninvasive imaging by combining near infrared light excitation and ultrasound detection. In this work, we develop the theoretical concept whereby the voltage-dependent quenching of dye fluorescence leads to a reciprocal enhancement of PA intensity. Based on this concept, we synthesized a novel near infrared photoacoustic VSD (PA-VSD) whose PA intensity change is sensitive to membrane potential. In the polarized state, this cyanine-based probe enhances PA intensity while decreasing fluorescence output in a lipid vesicle membrane model. With a 3-9 μM VSD concentration, we measured a PA signal increase in the range of 5.3 % to 18.1 %, and observed a corresponding signal reduction in fluorescence emission of 30.0 % to 48.7 %. A theoretical model successfully accounts for how the experimental PA intensity change depends on fluorescence and absorbance properties of the dye. These results not only demonstrate the voltage sensing capability of the dye, but also indicate the necessity of considering both fluorescence and absorbance spectral sensitivities in order to optimize the characteristics of improved photoacoustic probes. Together, our results demonstrate photoacoustic sensing as a potential new modality for sub-second recording and external imaging of electrophysiological and neurochemical events in the brain.
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