Electron-beam patterned calibration structures for structured illumination microscopy
S. Hari (TU Delft - ImPhys/Microscopy Instrumentation & Techniques)
Johan A. Slotman (Erasmus MC)
Y. Vos (TU Delft - ImPhys/Microscopy Instrumentation & Techniques)
Christian Floris (Student TU Delft)
W.A. van Cappellen (Erasmus MC)
C. W. Hagen (TU Delft - ImPhys/Microscopy Instrumentation & Techniques)
S. Stallinga (TU Delft - ImPhys/Imaging Physics)
A.B. Houtsmuller (Erasmus MC)
JP Hoogenboom (TU Delft - ImPhys/Microscopy Instrumentation & Techniques)
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Abstract
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.
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