Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging
Abstract Chemical imaging, especially mid-infrared spectroscopic microscopy, enables label-free biomedical analyses while achieving expansive molecular sensitivity. However, its slow speed and poor image quality impede widespread adoption. We present a microscope that provides high-throughput record...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-08-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-40740-w |
| _version_ | 1797558174183587840 |
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| author | Kevin Yeh Ishaan Sharma Kianoush Falahkheirkhah Matthew P. Confer Andres C. Orr Yen-Ting Liu Yamuna Phal Ruo-Jing Ho Manu Mehta Ankita Bhargava Wenyan Mei Georgina Cheng John C. Cheville Rohit Bhargava |
| author_facet | Kevin Yeh Ishaan Sharma Kianoush Falahkheirkhah Matthew P. Confer Andres C. Orr Yen-Ting Liu Yamuna Phal Ruo-Jing Ho Manu Mehta Ankita Bhargava Wenyan Mei Georgina Cheng John C. Cheville Rohit Bhargava |
| author_sort | Kevin Yeh |
| collection | DOAJ |
| description | Abstract Chemical imaging, especially mid-infrared spectroscopic microscopy, enables label-free biomedical analyses while achieving expansive molecular sensitivity. However, its slow speed and poor image quality impede widespread adoption. We present a microscope that provides high-throughput recording, low noise, and high spatial resolution where the bottom-up design of its optical train facilitates dual-axis galvo laser scanning of a diffraction-limited focal point over large areas using custom, compound, infinity-corrected refractive objectives. We demonstrate whole-slide, speckle-free imaging in ~3 min per discrete wavelength at 10× magnification (2 μm/pixel) and high-resolution capability with its 20× counterpart (1 μm/pixel), both offering spatial quality at theoretical limits while maintaining high signal-to-noise ratios (>100:1). The data quality enables applications of modern machine learning and capabilities not previously feasible – 3D reconstructions using serial sections, comprehensive assessments of whole model organisms, and histological assessments of disease in time comparable to clinical workflows. Distinct from conventional approaches that focus on morphological investigations or immunostaining techniques, this development makes label-free imaging of minimally processed tissue practical. |
| first_indexed | 2024-03-10T17:26:43Z |
| format | Article |
| id | doaj.art-b8115ce5b5b94b2bba79920b1d6e89cb |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-10T17:26:43Z |
| publishDate | 2023-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-b8115ce5b5b94b2bba79920b1d6e89cb2023-11-20T10:09:17ZengNature PortfolioNature Communications2041-17232023-08-0114111210.1038/s41467-023-40740-wInfrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imagingKevin Yeh0Ishaan Sharma1Kianoush Falahkheirkhah2Matthew P. Confer3Andres C. Orr4Yen-Ting Liu5Yamuna Phal6Ruo-Jing Ho7Manu Mehta8Ankita Bhargava9Wenyan Mei10Georgina Cheng11John C. Cheville12Rohit Bhargava13Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignBeckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignBeckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignBeckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignBeckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignBeckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignBeckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignBeckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignDepartment of Bioengineering, University of Illinois at Urbana-ChampaignUniversity of Illinois Laboratory High SchoolDepartment of Comparative Biosciences, College of Veterinary Medicine, University of Illinois at Urbana-ChampaignCancer Center at Illinois, University of Illinois at Urbana-ChampaignDepartment of Laboratory Medicine and Pathology, College of Medicine and Science, Mayo ClinicBeckman Institute for Advanced Science and Technology, University of Illinois at Urbana-ChampaignAbstract Chemical imaging, especially mid-infrared spectroscopic microscopy, enables label-free biomedical analyses while achieving expansive molecular sensitivity. However, its slow speed and poor image quality impede widespread adoption. We present a microscope that provides high-throughput recording, low noise, and high spatial resolution where the bottom-up design of its optical train facilitates dual-axis galvo laser scanning of a diffraction-limited focal point over large areas using custom, compound, infinity-corrected refractive objectives. We demonstrate whole-slide, speckle-free imaging in ~3 min per discrete wavelength at 10× magnification (2 μm/pixel) and high-resolution capability with its 20× counterpart (1 μm/pixel), both offering spatial quality at theoretical limits while maintaining high signal-to-noise ratios (>100:1). The data quality enables applications of modern machine learning and capabilities not previously feasible – 3D reconstructions using serial sections, comprehensive assessments of whole model organisms, and histological assessments of disease in time comparable to clinical workflows. Distinct from conventional approaches that focus on morphological investigations or immunostaining techniques, this development makes label-free imaging of minimally processed tissue practical.https://doi.org/10.1038/s41467-023-40740-w |
| spellingShingle | Kevin Yeh Ishaan Sharma Kianoush Falahkheirkhah Matthew P. Confer Andres C. Orr Yen-Ting Liu Yamuna Phal Ruo-Jing Ho Manu Mehta Ankita Bhargava Wenyan Mei Georgina Cheng John C. Cheville Rohit Bhargava Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging Nature Communications |
| title | Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging |
| title_full | Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging |
| title_fullStr | Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging |
| title_full_unstemmed | Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging |
| title_short | Infrared spectroscopic laser scanning confocal microscopy for whole-slide chemical imaging |
| title_sort | infrared spectroscopic laser scanning confocal microscopy for whole slide chemical imaging |
| url | https://doi.org/10.1038/s41467-023-40740-w |
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