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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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Detalles Bibliográficos
Autores principales: Hari, Sangeetha, Slotman, Johan A., Vos, Yoram, Floris, Christian, van Cappellen, Wiggert A., Hagen, C. W., Stallinga, Sjoerd, Houtsmuller, Adriaan B., Hoogenboom, Jacob P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9684522/
https://www.ncbi.nlm.nih.gov/pubmed/36418420
http://dx.doi.org/10.1038/s41598-022-24502-0
Descripción
Sumario: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.