Electron-beam patterned calibration structures for structured illumination microscopy

Electron-beam patterned calibration structures for structured illumination microscopy

Hari, S. (TU Delft ImPhys/Microscopy Instrumentation & Techniques)
Slotman, Johan A. (Erasmus MC)
Vos, Y. (TU Delft ImPhys/Microscopy Instrumentation & Techniques)
Floris, Christian (Student TU Delft)
van Cappellen, W.A. (Erasmus MC)
Hagen, C.W. (TU Delft ImPhys/Microscopy Instrumentation & Techniques)
Stallinga, S. (TU Delft ImPhys/Imaging Physics)
Houtsmuller, A.B. (Erasmus MC)
Hoogenboom, J.P. (TU Delft ImPhys/Microscopy Instrumentation & Techniques)

ImPhys/Imaging Physics

2022

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.

http://resolver.tudelft.nl/uuid:3ef49fdc-b935-4b4f-becd-992393abaa02

https://doi.org/10.1038/s41598-022-24502-0

2045-2322

Scientific Reports, 12 (1), 10

Institutional Repository

journal article

© 2022 S. Hari, Johan A. Slotman, Y. Vos, Christian Floris, W.A. van Cappellen, C.W. Hagen, S. Stallinga, A.B. Houtsmuller, J.P. Hoogenboom