Label-free intraoperative photoacoustic microscopy for rapid diagnosis of tissue biopsies with specimen integrity

摘要(30行) 术中通常需要冰冻切片诊断以指导手术。虽然这种方法论 仍然是不可替代的，它遭受着一系列的限制：组织丢失，冻结伪影，脂肪的次优切割 标本，无法诊断骨骼损伤，以及延长的周转时间。 其目的是开发一种多通道亚细胞分辨率光声显微镜(PAM)系统 解决所有上述限制，并为病理学家提供具有可比性的诊断图像 或与现有技术相同，具有匹配或超过标准透射光的放大倍数 显微镜，而且不需要对病理学家进行额外的培训即可诊断。 为了达到我们的目标，我们将追求以下两个具体目标： 具体目标1：开发多通道亚细胞分辨PAM系统，用于快速分析和诊断 组织活检 A.开发多通道亚细胞分辨率PAM系统 B.开发定制的组织活检固定器，用于PAM快速组织活检处理和成像 具体目标2：优化和验证多通道亚细胞分辨率PAM，以转换为强大的、 准确、高效的术中装置 优化图像采集，以最大限度地区分细胞核、细胞质和间质 组织降级最小的特性 B.优化采集后处理算法以生成类似经典苏木素的伪彩色图像 和伊红(H&E)切片，叠加独特的成像方式(水肿和胶原) C.通过与H&E切片进行比较，验证PAM生成的图像用于诊断目的的利用率。 它们是通过福尔马林固定的石蜡包埋(目前的黄金标准)制备的 D.通过与以下各项进行比较，验证PAM生成的图像是否可用于术中诊断 冰冻切片(现行的术中黄金标准) 影响：拟议的技术将是一项重大突破，解决了目前 术中以冰冻切片为基础的诊断，并提供具有H&E等效性的图像。它的目标是 介绍了一种快速、节省组织的手术活检诊断方法，对医疗保健产生了积极影响 系统和病人护理。减少冻结剖面解释的周转时间将显著 减少在手术室中花费的时间，从而降低患者风险和成本。有能力 评估小样本的充分性而不丢失组织将减少再次活组织检查的需要。 虚拟生成的图像可以远程解释，这样较小的医院就可以从手术中受益 目前仅适用于较大的三级医院和大学中心。

ABSTRACT (30 Lines) Frozen section diagnoses are typically requested intraoperatively to guide the surgery. While this methodology is still irreplaceable, it suffers from a series of limitations: tissue loss, freezing artifacts, suboptimal cutting of fatty specimens, inability to diagnose bony lesions, and prolonged turnaround time. The aim is to develop a multi-channel subcellular resolution photoacoustic microscopy (PAM) system capable of addressing all the aforementioned limitations and providing pathologists with diagnostic images comparable or identical to the state of the art, with magnification matching or exceeding that of standard transmission light microscopes, and requiring no additional training for pathologists to diagnose. To achieve our objective, we will pursue the following two specific aims: Specific Aim 1: Develop a multi-channel subcellular resolution PAM system for rapid analysis and diagnosis of tissue biopsies A. Develop a multi-channel subcellular resolution PAM system B. Develop a customized tissue biopsy holder for rapid tissue biopsy handling and imaging by PAM Specific Aim 2: Optimize and validate the multi-channel subcellular resolution PAM for translation into a robust, accurate, and efficient intraoperative device A. Optimize image acquisition in order to maximize discrimination of nuclear, cytoplasmic, and stromal properties with minimal tissue degradation B. Optimize post-acquisition algorithms to generate pseudocolored images resembling classical hematoxylin and eosin (H&E) sections with superimposed unique imaging modalities (edema and collagen) C. Validate the utilization of PAM generated images for diagnostic purposes by comparing with H&E sections, which are prepared by formalin-fixed paraffin-embedding (the current gold standard) D. Validate the utilization of PAM generated images for intraoperative diagnostic purposes by comparing with H&E sections, which are prepared by frozen section (the current intraoperative gold standard) Impact: The proposed technology will represent a major breakthrough by addressing all the limitations of current frozen section-based diagnoses and providing images with H&E equivalent qualities intraoperatively. It aims to introduce a rapid and tissue saving diagnostic method for surgical biopsies, positively impacting the healthcare system and patient care. Reducing the turnaround time on the frozen section interpretation will significantly reduce the time spent in the operating room, and therefore, decrease patient risks and costs. The ability to evaluate adequacy of small specimens with no loss in tissue will decrease the need for re-biopsy procedures. The virtually generated images can be interpreted remotely so that smaller hospitals can benefit from procedures currently available only to larger tertiary hospitals and university centers.