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However, the use of oil\u2010immersion 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\u2010immersion objectives also demands more sophisticated mechanical focusing. Here, we introduce a chromosome oil\u2010free microscopic imaging system based on Fourier Ptychographic Microscopy (FPM) technology. The system employs a 100\u00d7, 0.8 NA dry objective to achieve a half\u2010pitch resolution of 194\u2009nm at an incident wavelength of 524\u2009nm. The reconstructed images of chromosomes surpassed the conventional imaging with a 100\u00d7, 1.25 NA oil\u2010immersion objective. We also utilized digital refocusing methods to extend the effective DOF to \u00b12.4\u2009\u03bcm. This study preliminarily validates the possibility of developing a new generation of chromosome scanners without using an oil\u2010immersion objective.<\/jats:p>","DOI":"10.1002\/jbio.202400534","type":"journal-article","created":{"date-parts":[[2025,3,11]],"date-time":"2025-03-11T13:05:45Z","timestamp":1741698345000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["High\u2010Resolution Oil\u2010Free Imaging of Chromosomes Using Fourier Ptychographic Microscopy"],"prefix":"10.1002","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-6505-4062","authenticated-orcid":false,"given":"Changliang","family":"Yu","sequence":"first","affiliation":[{"name":"School of Biomedical Engineering (Suzhou), Division of Life Sciences and Medicine University of Science and Technology of China  Hefei China"},{"name":"CAS Key Lab of Bio\u2010Medical Diagnostics, Suzhou Institute of Biomedical Engineering and Technology 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