Electron-beam patterned calibration structures for structured illumination microscopy

Super-resolution fluorescence microscopy can be achieved by image reconstruction after spatially patterned illumination or sequential photo-switching and read-out. Reconstruction algorithms and microscope performance are typically tested using simulated image data, due to a lack of strategies to pat...

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Bibliographic Details
Published in:Scientific reports 2022-11, Vol.12 (1), p.20185-20185, Article 20185
Main Authors: Hari, Sangeetha, Slotman, Johan A., Vos, Yoram, Floris, Christian, van Cappellen, Wiggert A., Hagen, C. W., Stallinga, Sjoerd, Houtsmuller, Adriaan B., Hoogenboom, Jacob P.
Format: Article
Language:English
Subjects:
631/1647/328/2238
639/925/930/543
Algorithms
Calibration
Electrons
Humanities and Social Sciences
Lighting
Microscopy, Fluorescence
multidisciplinary
Science
Science (multidisciplinary)
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Summary:Super-resolution fluorescence microscopy can be achieved by image reconstruction after spatially patterned illumination or sequential photo-switching and read-out. Reconstruction algorithms and microscope performance are typically tested using simulated image data, due to a lack of strategies to pattern complex fluorescent patterns with nanoscale dimension control. Here, we report direct electron-beam patterning of fluorescence nanopatterns as calibration standards for super-resolution fluorescence. Patterned regions are identified with both electron microscopy and fluorescence labelling of choice, allowing precise correlation of predefined pattern dimensions, a posteriori obtained electron images, and reconstructed super-resolution images.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-022-24502-0