Background: Endodontic maladies incur significant costs to the military and are difficult to diagnose. Early identification and treatment of endodontic conditions are critical. Our research focused on designing a novel, vitality-based photoplethysmography imaging (PPGI) system capable of determining the pulse frequency within the dental pulp, allowing for direct, accurate, real-time visualization of tooth vitality. Objective: The objective of this study was to verify the accuracy of a developed video stabilization and digital signal processing algorithm – Pulp Assessment by Local Observation (PABLO). Methodology: MATLAB was used to create the PABLO video analysis algorithm to determine the pulse frequency of the dental pulp from PPGI signals. In order to verify the accuracy of this algorithm, four trials were conducted to test the algorithm under various parameters. The control group was simply a measurement of the algorithm’s ability to detect a pulse within an ex vivo tooth model. Following this, separate trials were conducted to test the effects of a simulated gumline, adjacent teeth, and a video stabilization protocol on the algorithm’s accuracy. Results: Video recordings from the ex vivo model were analyzed using the PABLO algorithm to determine its accuracy in detecting the pulse frequency of the pulp. Results of the analysis showed that the algorithm had a pulse detection sensitivity above 90% and a percent error less than 11% in all trials. In the control case, results showed a sensitivity of 93% and a pulse detection error of 7.3%, indicating that this algorithm has promise as a diagnostic tool for clinicians.
To study cancer progression and drug response, researchers have developed methods to derive tumor organoids from primary tissues, which not only have the same proteomic and genetic abnormalities as the malignant disease but also better replicate tumor behaviors than 2-dimensional culture models. It has been shown that tumor organoids can be used to predict treatment response, understand drug resistance, and study tumor heterogeneity at the individual-patient level. Whereas large-scale production of patient-derived organoids in standard flat-bottom 1,536-well plates has recently been demonstrated for cytotoxicity screening of 3,300 approved drugs, no suitable functional imaging tool can provide rapid 3-dimensional (3D) evaluation over a wide range of cellular states in these mass-produced organoids. The traditional two-photon scanning microscopy is too slow, while the conventional light-sheet microscopy is not compatible with microwell plates. Here we propose a microplate-compatible single-objective multiphoton light-sheet microscopy (SOMP- LSM) that can provide high imaging speed for 3D imaging analysis of organoids and deeper imaging depth with sub-cellular resolution. Our simulation of 3D point-spread function simulation for the SO-MP-LSM shows that our imaging system can achieve 270 nm lateral resolution and 800 nm axial resolution deep into organoids.
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