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STED‐TEM Correlative Microscopy Leveraging Nanodiamonds as Intracellular Dual‐Contrast Markers
Author(s) -
Prabhakar Neeraj,
Peurla Markus,
Koho Sami,
Deguchi Takahiro,
Näreoja Tuomas,
Chang HuanCheng,
Rosenholm Jessica M.,
Hänninen Pekka E.
Publication year - 2018
Publication title -
small
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.785
H-Index - 236
eISSN - 1613-6829
pISSN - 1613-6810
DOI - 10.1002/smll.201701807
Subject(s) - sted microscopy , microscopy , confocal , correlative , materials science , fluorescence , transmission electron microscopy , fluorescence lifetime imaging microscopy , confocal microscopy , live cell imaging , fluorescence microscope , microscope , electron microscope , optics , nanotechnology , stimulated emission , chemistry , physics , laser , cell , linguistics , philosophy , biochemistry
Development of fluorescent and electron dense markers is essential for the implementation of correlative light and electron microscopy, as dual‐contrast landmarks are required to match the details in the multimodal images. Here, a novel method for correlative microscopy that utilizes fluorescent nanodiamonds (FNDs) as dual‐contrast probes is reported. It is demonstrated how the FNDs can be used as dual‐contrast labels—and together with automatic image registration tool SuperTomo, for precise image correlation—in high‐resolution stimulated emission depletion (STED)/confocal and transmission electron microscopy (TEM) correlative microscopy experiments. It is shown how FNDs can be employed in experiments with both live and fixed cells as well as simple test samples. The fluorescence imaging can be performed either before TEM imaging or after, as the robust FNDs survive the TEM sample preparation and can be imaged with STED and other fluorescence microscopes directly on the TEM grids.