Author:
Hari Sangeetha,Slotman Johan A.,Vos Yoram,Floris Christian,van Cappellen Wiggert A.,Hagen C. W.,Stallinga Sjoerd,Houtsmuller Adriaan B.,Hoogenboom Jacob P.
Abstract
AbstractSuper-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.
Funder
NanoNextNL, a consortium of the Dutch government and 130 public and private partners.
ZonMw
Nederlandse Organisatie voor Wetenschappelijk Onderzoek
Publisher
Springer Science and Business Media LLC