Impact of uniform illumination in widefield microscopy and mesoscopy
Abstract Illumination uniformity is critical for widefield optical microscopy, especially for high-throughput and accurate quantitative imaging of biological specimens. While traditional Köhler illumination improves uniformity, it often fails to deliver homogeneous intensity across large fields of v...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-07-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-025-08754-0 |
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| author | Mete Aslan Alexander J. Gray Logan Packard Caeden Giorgetta Irving J. Bigio M. Selim Ünlü |
| author_facet | Mete Aslan Alexander J. Gray Logan Packard Caeden Giorgetta Irving J. Bigio M. Selim Ünlü |
| author_sort | Mete Aslan |
| collection | DOAJ |
| description | Abstract Illumination uniformity is critical for widefield optical microscopy, especially for high-throughput and accurate quantitative imaging of biological specimens. While traditional Köhler illumination improves uniformity, it often fails to deliver homogeneous intensity across large fields of view. Existing optical and computational correction techniques remain inadequate for a broad range of quantitative imaging applications. Here, we implement a novel illumination device that we call the “effective uniform color-light integration device” (EUCLID), quantifying improvements in two widefield imaging modalities that require uniform illumination. For both imaging modalities, we demonstrate significantly improved precision of quantitative measurements compared to traditional Köhler illumination. The EUCLID device, which can also provide uniform spectral mixing, can be readily adapted to many other widefield imaging modalities to enhance imaging accuracy and reliability, with low cost and ease of implementation. |
| format | Article |
| id | doaj-art-3b1fb16caabc43e09db78530b7be99c8 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-3b1fb16caabc43e09db78530b7be99c82025-08-20T03:45:52ZengNature PortfolioScientific Reports2045-23222025-07-0115111210.1038/s41598-025-08754-0Impact of uniform illumination in widefield microscopy and mesoscopyMete Aslan0Alexander J. Gray1Logan Packard2Caeden Giorgetta3Irving J. Bigio4M. Selim Ünlü5Department of Electrical and Computer Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Biomedical Engineering, Boston UniversityDepartment of Mechanical Engineering, Boston UniversityDepartment of Electrical and Computer Engineering, Boston UniversityDepartment of Electrical and Computer Engineering, Boston UniversityAbstract Illumination uniformity is critical for widefield optical microscopy, especially for high-throughput and accurate quantitative imaging of biological specimens. While traditional Köhler illumination improves uniformity, it often fails to deliver homogeneous intensity across large fields of view. Existing optical and computational correction techniques remain inadequate for a broad range of quantitative imaging applications. Here, we implement a novel illumination device that we call the “effective uniform color-light integration device” (EUCLID), quantifying improvements in two widefield imaging modalities that require uniform illumination. For both imaging modalities, we demonstrate significantly improved precision of quantitative measurements compared to traditional Köhler illumination. The EUCLID device, which can also provide uniform spectral mixing, can be readily adapted to many other widefield imaging modalities to enhance imaging accuracy and reliability, with low cost and ease of implementation.https://doi.org/10.1038/s41598-025-08754-0Optical microscopyWide-field imagingQuantitative imagingMesoscopyIllumination uniformityWhole slide scanning |
| spellingShingle | Mete Aslan Alexander J. Gray Logan Packard Caeden Giorgetta Irving J. Bigio M. Selim Ünlü Impact of uniform illumination in widefield microscopy and mesoscopy Scientific Reports Optical microscopy Wide-field imaging Quantitative imaging Mesoscopy Illumination uniformity Whole slide scanning |
| title | Impact of uniform illumination in widefield microscopy and mesoscopy |
| title_full | Impact of uniform illumination in widefield microscopy and mesoscopy |
| title_fullStr | Impact of uniform illumination in widefield microscopy and mesoscopy |
| title_full_unstemmed | Impact of uniform illumination in widefield microscopy and mesoscopy |
| title_short | Impact of uniform illumination in widefield microscopy and mesoscopy |
| title_sort | impact of uniform illumination in widefield microscopy and mesoscopy |
| topic | Optical microscopy Wide-field imaging Quantitative imaging Mesoscopy Illumination uniformity Whole slide scanning |
| url | https://doi.org/10.1038/s41598-025-08754-0 |
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