Label-free digital cytopathology using deep-ultraviolet coded ptychography with intrinsic molecular contrast

项目摘要/摘要 组织病理学检查是现代医学和生物学研究的支柱。通常 使用明视场显微镜需要事先准备安装在玻璃上的微米厚的组织切片 幻灯片和染料染色，这一过程需要数小时或数天，延迟了对关键信息的访问 干预措施。本项目旨在开发一种新型的高通量数字细胞病理学工具， 无标记和无载玻片组织学的固有分子对比。该工具将基于无透镜技术构建 高通量编码光刻技术在260 nm和280 nm深紫外光(DUV)下工作 波长。DUV光直接与核酸(核)生物分子相互作用并被其吸收 细胞中的氨基酸(蛋白质)。这两种生物分子的吸收光谱是不同的。通过 在260 nm和280 nm的DUV波长测量样品的剖面，我们可以恢复 不需要任何染料标记的细胞核和蛋白质的定量质谱图。相比之下，没有 这些生物分子在可见光区域的吸收。 在所提出的DUV编码的光刻工具中，我们不像在传统的光刻工具中那样使用任何光学透镜 显微镜平台。取而代之的是，我们在图像传感器的顶部制造了一个无序工程编码层。 这个编码层充当了一个大规模的散射镜，用于对放置在其上的样本进行成像。什么时候 光从组织样品大角度衍射，编码层将衍射光重定向到 传感器像素可以检测到的较小角度。因此，原本无法访问的高分辨率 可以使用编码层下面的像素阵列来获取对象细节。我们的初步数据显示 图像吞吐量超过了世界上最快的全幻灯片扫描仪：分辨率为308纳米 线宽可以在15秒内覆盖240毫米的有效视野。通过使用双波长DUV LED用于 样本照明，所提出的工具将提供基于相位的定量形态测量 以及基于幅度的不同生物分子的固有化学特性。可用于快速现场 细针抽吸细胞学涂片的评价。它还可以减少样品准备时间 并提供手术切缘的术中病理指导。整个设备的成本将是 与iPad类似，使其成为大多数人手持的、高吞吐量的完整幻灯片扫描仪 世界各地的个别病理学家和研究人员。

Project Summary / Abstract Histopathological examination of tissue is a pillar of modern medicine and biological research. Commonly used bright-field microscopy requires prior preparation of micrometer-thick tissue sections mounted on glass slides and stained with dyes, a process that takes hours or days, delaying access to critical information for interventions. This project aims to develop a new type of high-throughput digital cytopathology tool with intrinsic molecular contrast for label-free and slide-free histology. This tool will be built based on lensless high-throughput coded ptychography operated at the 260-nm and 280-nm deep ultraviolet (DUV) wavelengths. DUV light directly interacts with and is absorbed by biomolecules of nucleic acids (nucleus) and amino acids (protein) of cells. Absorption spectra are different for these two types of biomolecules. By measuring the specimen profiles at the 260-nm and 280-nm DUV wavelengths, we can recover the quantitative mass maps of cell nucleus and protein without any dye labeling. In contrast, there is no absorption for these biomolecules in the visible light regime. In the proposed DUV coded ptychography tool, we do not use any optical lens as in a conventional microscope platform. Instead, we fabricate a disorder-engineered coded layer on top of an image sensor. This coded layer serves as a large-scale scattering ‘lens’ for imaging specimens placed on top of it. When light diffracts from the tissue samples at a large angle, the coded layer redirects the diffracted light into smaller angles that are detectable by sensor pixels. As such, the otherwise inaccessible high-resolution object details can be acquired using the pixel array underneath the coded layer. Our preliminary data show that the image throughput is greater than the fastest whole slide scanner in the world: resolving 308-nm linewidth over a 240-mm2 effective field of view in 15 seconds. By using the dual wavelength DUV LEDs for sample illumination, the proposed tool will provide both phase-based quantitative morphology measurement and amplitude-based intrinsic chemical specificities for different biomolecules. It can be used for rapid onsite evaluation of cytology smear obtained from fine-needle aspirate. It can also reduce sample preparation time and provides intraoperative pathology guidance on surgical margins. The cost of the entire device will be similar to that of an iPad, making it a handheld, high-throughput ‘personal’ whole slide scanner for most individual pathologists and researchers worldwide.