From the Department of Laboratory Medicine and Pathology (Reddi, Liu), University of Washington, Seattle.
Department of Mechanical Engineering (Barner, Gao, Liu), University of Washington, Seattle.
Arch Pathol Lab Med. 2023 Oct 1;147(10):1164-1171. doi: 10.5858/arpa.2022-0133-OA.
CONTEXT.—: Anatomic pathologists render diagnosis on tissue samples sectioned onto glass slides and viewed under a bright-field microscope. This approach is destructive to the sample, which can limit its use for ancillary assays that can inform patient management. Furthermore, the subjective interpretation of a relatively small number of 2D tissue sections per sample contributes to low interobserver agreement among pathologists for the assessment (diagnosis and grading) of various lesions.
OBJECTIVE.—: To evaluate 3D pathology data sets of thick formalin-fixed Barrett esophagus specimens imaged nondestructively with open-top light-sheet (OTLS) microscopy.
DESIGN.—: Formalin-fixed, paraffin-embedded Barrett esophagus samples (N = 15) were deparaffinized, stained with a fluorescent analog of hematoxylin-eosin, optically cleared, and imaged nondestructively with OTLS microscopy. The OTLS microscopy images were subsequently compared with archived hematoxylin-eosin histology sections from each sample.
RESULTS.—: Barrett esophagus samples, both small endoscopic forceps biopsies and endoscopic mucosal resections, exhibited similar resolvable structures between OTLS microscopy and conventional light microscopy with up to a ×20 objective (×200 overall magnification). The 3D histologic images generated by OTLS microscopy can enable improved discrimination of cribriform and well-formed gland morphologies. In addition, a much larger amount of tissue is visualized with OTLS microscopy, which enables improved assessment of clinical specimens exhibiting high spatial heterogeneity.
CONCLUSIONS.—: In esophageal specimens, OTLS microscopy can generate images comparable in quality to conventional light microscopy, with the advantages of providing 3D information for enhanced evaluation of glandular morphologies and enabling much more of the tissue specimen to be visualized nondestructively.
解剖病理学家对切成玻璃片的组织样本进行诊断，并在明场显微镜下观察。这种方法对样本具有破坏性，这可能限制了其用于辅助检测的应用，而这些辅助检测可以为患者管理提供信息。此外，由于每个样本中只有少数 2D 组织切片被主观解释，因此病理学家对各种病变的评估（诊断和分级）的观察者间一致性较低。
评估经非破坏性开放式顶片光片（OTLS）显微镜成像的厚福尔马林固定 Barrett 食管标本的 3D 病理学数据集。
福尔马林固定、石蜡包埋的 Barrett 食管样本（N=15）被脱石蜡、用苏木精-伊红的荧光类似物染色、光学透明化，并经 OTLS 显微镜进行非破坏性成像。随后将 OTLS 显微镜图像与每个样本的存档苏木精-伊红组织学切片进行比较。
Barrett 食管样本，包括小内镜活检钳活检和内镜黏膜切除术，在 OTLS 显微镜和传统的光学显微镜之间显示出相似的可分辨结构，最高可达×20 物镜（总放大倍数×200）。OTLS 显微镜生成的 3D 组织学图像可以提高对筛状和形态良好的腺体形态的区分能力。此外，OTLS 显微镜可以可视化更多的组织，这可以改善对具有高空间异质性的临床标本的评估。
在食管标本中，OTLS 显微镜可以生成与传统光学显微镜质量相当的图像，其优势在于提供 3D 信息，以增强对腺体形态的评估，并使更多的组织标本可以进行非破坏性可视化。
