Mori Nobuhito, Miyazaki Yoshihiro, Oda Tatsuya, Kida Yasuyuki S
Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba 305-8565, Ibaraki, Japan.
Department of Gastrointestinal and Hepato-Biliary-Pancreatic Surgery, Institute of Medicine, University of Tsukuba, Tsukuba 305-8575, Ibaraki, Japan.
Sensors (Basel). 2025 May 8;25(10):2968. doi: 10.3390/s25102968.
The creation of virtual slides, i.e., high-resolution digital images of biological samples, is expensive, and existing manual methods often suffer from stitching errors and additional reimaging costs. To address these issues, we propose a real-time mosaicing manager for manual microscopy (RT-4M) that performs real-time stitching and allows users to preview slides during imaging using existing manual microscopy systems, thereby reducing the need for reimaging. We install it on two different microscopy systems, successfully creating virtual slides of hematoxylin and eosin- and fluorescent-stained tissues obtained from humans and mice. The fluorescent-stained tissues consist of two colors, requiring the manual switching of the filter and an exposure time of 1.6 s per color. Even in the case of the largest dataset in this study (over 900 images), the entire sample is captured without any omissions. Moreover, RT-4M exhibits a processing time of less than one second per registration, indicating that it does not hinder the user's imaging workflow. Additionally, the composition process reduces the misalignment rate by a factor of 20 compared to existing software. We believe that the proposed software will prove useful in the fields of pathology and bio-research, particularly for facilities with relatively limited budgets.
虚拟玻片的创建，即生物样本的高分辨率数字图像，成本高昂，而且现有的手动方法常常存在拼接错误以及额外的重新成像成本。为了解决这些问题，我们提出了一种用于手动显微镜的实时拼接管理器（RT-4M），它能进行实时拼接，并允许用户在使用现有的手动显微镜系统成像过程中预览玻片，从而减少重新成像的需求。我们将其安装在两种不同的显微镜系统上，成功创建了从人和小鼠获取的苏木精和伊红染色以及荧光染色组织的虚拟玻片。荧光染色组织由两种颜色组成，需要手动切换滤光片，且每种颜色的曝光时间为1.6秒。即使在本研究中最大的数据集（超过900张图像）的情况下，整个样本也能毫无遗漏地被捕获。此外，RT-4M每次配准的处理时间不到一秒，这表明它不会妨碍用户的成像工作流程。此外，与现有软件相比，合成过程将未对准率降低了20倍。我们相信，所提出的软件将在病理学和生物研究领域证明是有用的，特别是对于预算相对有限的机构。
