{"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":"<p><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>","PeriodicalId":94068,"journal":{"name":"Journal of biophotonics","volume":" ","pages":"e202400534"},"PeriodicalIF":0.0000,"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":"1085","ListUrlMain":"https://doi.org/10.1002/jbio.202400534","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
Abstract
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.