{"title":"无监督学习可实现多色同步荧光显微镜观察,且无细胞结构串扰","authors":"Bolin Lu, Zhangheng Ding, Kefu Ning, Xiaoyu Zhang, Xiangning Li, Jiangjiang Zhao, Ruiheng Xie, Dan Shen, Jiahong Hu, Tao Jiang, Jianwei Chen, Hui Gong, Jing Yuan","doi":"10.1063/5.0202622","DOIUrl":null,"url":null,"abstract":"In multicolor fluorescence microscopy, it is crucial to orient biological structures at a single-cell resolution based on precise anatomical annotations of cytoarchitecture images. However, during synchronous multicolor imaging, due to spectral mixing, the crosstalk from the blue signals of 4′,6-diamidino-2-phenylindole (DAPI)-stained cytoarchitecture images to the green waveband hinders the visualization and identification of green signals. Here, we proposed a deep learning-based framework named the crosstalk elimination and cytoarchitecture enhancement pipeline (CECEP) to simultaneously acquire crosstalk-free signals in the green channel and high-contrast DAPI-stained cytoarchitecture images during multicolor fluorescence imaging. For the CECEP network, we proposed an unsupervised learning algorithm named the cytoarchitecture enhancement network (CENet), which increased the signal-to-background ratio (SBR) of the cytoarchitecture images from 1.5 to 15.0 at a reconstruction speed of 25 Hz for 1800 × 1800 pixel images. The CECEP network is widely applicable to images of different quality, different types of tissues, and different multicolor fluorescence microscopy. In addition, the CECEP network can also facilitate various downstream analysis tasks, such as cell recognition, structure tensor calculation, and brain region segmentation. With the CECEP network, we simultaneously acquired two specific fluorescence-labeled neuronal distributions and their colocated high-SBR cytoarchitecture images without crosstalk throughout the brain. Experimental results demonstrate that our method could potentially facilitate multicolor fluorescence imaging applications in biology, such as revealing and visualizing different types of biological structures with precise locations and orientations.","PeriodicalId":8198,"journal":{"name":"APL Photonics","volume":"28 1","pages":""},"PeriodicalIF":5.4000,"publicationDate":"2024-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Unsupervised learning enables multicolor synchronous fluorescence microscopy without cytoarchitecture crosstalk\",\"authors\":\"Bolin Lu, Zhangheng Ding, Kefu Ning, Xiaoyu Zhang, Xiangning Li, Jiangjiang Zhao, Ruiheng Xie, Dan Shen, Jiahong Hu, Tao Jiang, Jianwei Chen, Hui Gong, Jing Yuan\",\"doi\":\"10.1063/5.0202622\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"In multicolor fluorescence microscopy, it is crucial to orient biological structures at a single-cell resolution based on precise anatomical annotations of cytoarchitecture images. However, during synchronous multicolor imaging, due to spectral mixing, the crosstalk from the blue signals of 4′,6-diamidino-2-phenylindole (DAPI)-stained cytoarchitecture images to the green waveband hinders the visualization and identification of green signals. Here, we proposed a deep learning-based framework named the crosstalk elimination and cytoarchitecture enhancement pipeline (CECEP) to simultaneously acquire crosstalk-free signals in the green channel and high-contrast DAPI-stained cytoarchitecture images during multicolor fluorescence imaging. For the CECEP network, we proposed an unsupervised learning algorithm named the cytoarchitecture enhancement network (CENet), which increased the signal-to-background ratio (SBR) of the cytoarchitecture images from 1.5 to 15.0 at a reconstruction speed of 25 Hz for 1800 × 1800 pixel images. The CECEP network is widely applicable to images of different quality, different types of tissues, and different multicolor fluorescence microscopy. In addition, the CECEP network can also facilitate various downstream analysis tasks, such as cell recognition, structure tensor calculation, and brain region segmentation. With the CECEP network, we simultaneously acquired two specific fluorescence-labeled neuronal distributions and their colocated high-SBR cytoarchitecture images without crosstalk throughout the brain. Experimental results demonstrate that our method could potentially facilitate multicolor fluorescence imaging applications in biology, such as revealing and visualizing different types of biological structures with precise locations and orientations.\",\"PeriodicalId\":8198,\"journal\":{\"name\":\"APL Photonics\",\"volume\":\"28 1\",\"pages\":\"\"},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2024-05-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"APL Photonics\",\"FirstCategoryId\":\"101\",\"ListUrlMain\":\"https://doi.org/10.1063/5.0202622\",\"RegionNum\":1,\"RegionCategory\":\"物理与天体物理\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"OPTICS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"APL Photonics","FirstCategoryId":"101","ListUrlMain":"https://doi.org/10.1063/5.0202622","RegionNum":1,"RegionCategory":"物理与天体物理","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"OPTICS","Score":null,"Total":0}
Unsupervised learning enables multicolor synchronous fluorescence microscopy without cytoarchitecture crosstalk
In multicolor fluorescence microscopy, it is crucial to orient biological structures at a single-cell resolution based on precise anatomical annotations of cytoarchitecture images. However, during synchronous multicolor imaging, due to spectral mixing, the crosstalk from the blue signals of 4′,6-diamidino-2-phenylindole (DAPI)-stained cytoarchitecture images to the green waveband hinders the visualization and identification of green signals. Here, we proposed a deep learning-based framework named the crosstalk elimination and cytoarchitecture enhancement pipeline (CECEP) to simultaneously acquire crosstalk-free signals in the green channel and high-contrast DAPI-stained cytoarchitecture images during multicolor fluorescence imaging. For the CECEP network, we proposed an unsupervised learning algorithm named the cytoarchitecture enhancement network (CENet), which increased the signal-to-background ratio (SBR) of the cytoarchitecture images from 1.5 to 15.0 at a reconstruction speed of 25 Hz for 1800 × 1800 pixel images. The CECEP network is widely applicable to images of different quality, different types of tissues, and different multicolor fluorescence microscopy. In addition, the CECEP network can also facilitate various downstream analysis tasks, such as cell recognition, structure tensor calculation, and brain region segmentation. With the CECEP network, we simultaneously acquired two specific fluorescence-labeled neuronal distributions and their colocated high-SBR cytoarchitecture images without crosstalk throughout the brain. Experimental results demonstrate that our method could potentially facilitate multicolor fluorescence imaging applications in biology, such as revealing and visualizing different types of biological structures with precise locations and orientations.
APL PhotonicsPhysics and Astronomy-Atomic and Molecular Physics, and Optics
CiteScore
10.30
自引率
3.60%
发文量
107
审稿时长
19 weeks
期刊介绍:
APL Photonics is the new dedicated home for open access multidisciplinary research from and for the photonics community. The journal publishes fundamental and applied results that significantly advance the knowledge in photonics across physics, chemistry, biology and materials science.