Electron-beam patterned calibration structures for structured illumination microscopy

Sangeetha Hari, Johan A. Slotman, Yoram Vos, Christian Floris, Wiggert A. van Cappellen, C. W. Hagen, Sjoerd Stallinga, Adriaan B. Houtsmuller, Jacob P. Hoogenboom*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

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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.

Original languageEnglish
Article number20185
Pages (from-to)10
JournalScientific Reports
Issue number1
Publication statusPublished - 2022


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