For the 1.7 million patients per year in the U.S. who receive a new cancer diagnosis, treatment decisions are largely made after a histopathology exam. Unfortunately, the gold standard of slide-based microscopic pathology suffers from high inter-observer variability and limited prognostic value due to sampling limitations and the inability to visualize tissue structures and molecular targets in their native 3D context. Here, we show that an open-top light-sheet microscope optimized for non-destructive slide-free pathology of clinical specimens enables the rapid imaging of intact tissues at high resolution over large 2D and 3D fields of view, with the same level of detail as traditional pathology. We demonstrate the utility of this technology for various applications: wide-area surface microscopy to triage surgical specimens (with ~200 μm surface irregularities), rapid intraoperative assessment of tumour-margin surfaces (12.5 sec/cm2), and volumetric assessment of optically cleared core–needle biopsies (1 mm in diameter, 2 cm in length). Light-sheet microscopy can be a versatile tool for both rapid surface microscopy and deep volumetric microscopy of human specimens.

Recent studies have proposed that light emitted by the Cherenkov effect may be used for a number of radiation therapy dosimetry applications. There is a correlation between the captured light and expected dose under certain conditions, yet discrepancies have also been observed and a complete examination of the theoretical differences has not been done. In this study, a fundamental comparison between the Cherenkov emission and absorbed dose was explored for x-ray photons, electrons, and protons using both a theoretical and Monte Carlo-based analysis. Based on the findings of where dose correlates with Cherenkov emission, it was concluded that for x-ray photons the light emission would be optimally suited for narrow beam stereotactic radiation therapy and surgery validation studies, for verification of dynamic intensity-modulated and volumetric modulated arc therapy treatment plans in water tanks, near monoenergetic sources (e.g., Co-60 and brachy therapy sources) and also for entrance and exit surface imaging dosimetry of both narrow and broad beams. For electron use, Cherenkov emission was found to be only suitable for surface dosimetry applications. Finally, for proton dosimetry, there exists a fundamental lack of Cherenkov emission at the Bragg peak, making the technique of little use, although post-irradiation detection of light emission from radioisotopes could prove to be useful.

Purpose: A novel technique for beam profiling of megavoltage photon beams was investigated for the first time by capturing images of the inducedČerenkov emission in water, as a potential surrogate for the imparted dose in irradiated media. Methods: A high-sensitivity, intensified CCD camera (ICCD) was configured to acquire 2D projection images ofČerenkov emission from a 4 × 4 cm 2 6 MV linear accelerator (LINAC) x-ray photon beam operating at a dose rate of 400 MU/min incident on a water tank with transparent walls. The ICCD acquisition was gated to the LINAC sync pulse to reduce background light artifacts, and the measurement quality was investigated by evaluating the signal to noise ratio and measurement repeatability as a function of delivered dose. Monte Carlo simulations were used to derive a calibration factor for differences between the optical images and deposited dose arising from the anisotropic angular dependence ofČerenkov emission. Finally,Čerenkov-based beam profiles were compared to a percent depth dose (PDD) and lateral dose profile at a depth of d max from a reference dose distribution generated from the clinical Varian ECLIPSE treatment planning system (TPS). Results: The signal to noise ratio was found to be 20 at a delivered dose of 66.6 cGy, and proportional to the square root of the delivered dose as expected from Poisson photon counting statistics. A 2.1% mean standard deviation and 5.6% maximum variation in successive measurements were observed, and the Monte Carlo derived calibration factor resulted inČerenkov emission images which were directly correlated to deposited dose, with some spatial issues. The dose difference between the TPS and PDD predicted byČerenkov measurements was within 20% in the buildup region with a distance to agreement (DTA) of 1.5-2 mm and ±3% at depths beyond d max . In the lateral profile, the dose difference at the beam penumbra was within ±13% with a DTA of 0-2 mm, ±5% in the central beam region, and 2%-3% in the beam umbra. Conclusions:The results from this initial study demonstrate the first documented use ofČerenkov emission imaging to profile x-ray photon LINAC beams in water. The proposed modality has several potential advantages over alternative methods, and upon future refinement may prove to be a robust and novel dosimetry method.

Recent advances in optical clearing and light-sheet microscopy have provided unprecedented access to structural and molecular information from intact tissues. However, current light-sheet microscopes have imposed constraints on the size, shape, number of specimens, and compatibility with various clearing protocols. Here we present a multi-immersion open-top light-sheet microscope that enables simple mounting of multiple specimens processed with a variety of clearing protocols, which will facilitate wide adoption by preclinical researchers and clinical laboratories. In particular, the open-top geometry provides unsurpassed versatility to interface with a wide range of accessory technologies in the future.