Journal
JOURNAL OF MICROSCOPY
Volume 252, Issue 1, Pages 58-70Publisher
WILEY
DOI: 10.1111/jmi.12071
Keywords
correlative microscopy; light microscopy; scanning electron microscopy; cellular imaging; Cathodoluminescence microscopy; fluorescence microscopy
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Funding
- STW Valorization Grant
- NanoNextNL
- nanotechnology consortium of the Government of the Netherlands
- 130 partners
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Summary We present an integrated light-electron microscope in which an inverted high-NA objective lens is positioned inside a scanning electron microscope (SEM). The SEM objective lens and the light objective lens have a common axis and focal plane, allowing high-resolution optical microscopy and scanning electron microscopy on the same area of a sample simultaneously. Components for light illumination and detection can be mounted outside the vacuum, enabling flexibility in the construction of the light microscope. The light objective lens can be positioned underneath the SEM objective lens during operation for sub-10 mu m alignment of the fields of view of the light and electron microscopes. We demonstrate in situ epifluorescence microscopy in the SEM with a numerical aperture of 1.4 using vacuum-compatible immersion oil. For a 40-nm-diameter fluorescent polymer nanoparticle, an intensity profile with a FWHM of 380 nm is measured whereas the SEM performance is uncompromised. The integrated instrument may offer new possibilities for correlative light and electron microscopy in the life sciences as well as in physics and chemistry.
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