{"title":"利用傅立叶平片显微镜对染色体进行高分辨率无油成像。","authors":"Changliang Yu, Yuguo Tang","doi":"10.1002/jbio.202400534","DOIUrl":null,"url":null,"abstract":"<div>\n \n <p>In conventional microscopic imaging of chromosomes, the use of high numerical aperture (NA) oil-immersion objectives is essential. However, the use of oil-immersion objectives poses a significant challenge to automated imaging systems because it increases the risk of sample contamination and instrumental damage. The shallow depth of field (DOF) of oil-immersion objectives also demands more sophisticated mechanical focusing. Here, we introduce a chromosome oil-free microscopic imaging system based on Fourier Ptychographic Microscopy (FPM) technology. The system employs a 100×, 0.8 NA dry objective to achieve a half-pitch resolution of 194 nm at an incident wavelength of 524 nm. The reconstructed images of chromosomes surpassed the conventional imaging with a 100×, 1.25 NA oil-immersion objective. We also utilized digital refocusing methods to extend the effective DOF to ±2.4 μm. This study preliminarily validates the possibility of developing a new generation of chromosome scanners without using an oil-immersion objective.</p>\n </div>","PeriodicalId":184,"journal":{"name":"Journal of Biophotonics","volume":"18 5","pages":""},"PeriodicalIF":2.0000,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"High-Resolution Oil-Free Imaging of Chromosomes Using Fourier Ptychographic Microscopy\",\"authors\":\"Changliang Yu, Yuguo Tang\",\"doi\":\"10.1002/jbio.202400534\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n <p>In conventional microscopic imaging of chromosomes, the use of high numerical aperture (NA) oil-immersion objectives is essential. However, the use of oil-immersion objectives poses a significant challenge to automated imaging systems because it increases the risk of sample contamination and instrumental damage. The shallow depth of field (DOF) of oil-immersion objectives also demands more sophisticated mechanical focusing. Here, we introduce a chromosome oil-free microscopic imaging system based on Fourier Ptychographic Microscopy (FPM) technology. The system employs a 100×, 0.8 NA dry objective to achieve a half-pitch resolution of 194 nm at an incident wavelength of 524 nm. The reconstructed images of chromosomes surpassed the conventional imaging with a 100×, 1.25 NA oil-immersion objective. We also utilized digital refocusing methods to extend the effective DOF to ±2.4 μm. This study preliminarily validates the possibility of developing a new generation of chromosome scanners without using an oil-immersion objective.</p>\\n </div>\",\"PeriodicalId\":184,\"journal\":{\"name\":\"Journal of Biophotonics\",\"volume\":\"18 5\",\"pages\":\"\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2025-02-24\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Biophotonics\",\"FirstCategoryId\":\"101\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/jbio.202400534\",\"RegionNum\":3,\"RegionCategory\":\"物理与天体物理\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Biophotonics","FirstCategoryId":"101","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/jbio.202400534","RegionNum":3,"RegionCategory":"物理与天体物理","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
High-Resolution Oil-Free Imaging of Chromosomes Using Fourier Ptychographic Microscopy
In conventional microscopic imaging of chromosomes, the use of high numerical aperture (NA) oil-immersion objectives is essential. However, the use of oil-immersion objectives poses a significant challenge to automated imaging systems because it increases the risk of sample contamination and instrumental damage. The shallow depth of field (DOF) of oil-immersion objectives also demands more sophisticated mechanical focusing. Here, we introduce a chromosome oil-free microscopic imaging system based on Fourier Ptychographic Microscopy (FPM) technology. The system employs a 100×, 0.8 NA dry objective to achieve a half-pitch resolution of 194 nm at an incident wavelength of 524 nm. The reconstructed images of chromosomes surpassed the conventional imaging with a 100×, 1.25 NA oil-immersion objective. We also utilized digital refocusing methods to extend the effective DOF to ±2.4 μm. This study preliminarily validates the possibility of developing a new generation of chromosome scanners without using an oil-immersion objective.
期刊介绍:
The first international journal dedicated to publishing reviews and original articles from this exciting field, the Journal of Biophotonics covers the broad range of research on interactions between light and biological material. The journal offers a platform where the physicist communicates with the biologist and where the clinical practitioner learns about the latest tools for the diagnosis of diseases. As such, the journal is highly interdisciplinary, publishing cutting edge research in the fields of life sciences, medicine, physics, chemistry, and engineering. The coverage extends from fundamental research to specific developments, while also including the latest applications.