SIM0216

ELECTRON MICROSCOPY
Integrated Raman – FIB – SEM
A Correlative Light and Electron Microscopy Study
Frank Timmermans 1 ,Barbara Liszka 1 ,Derya Ataç 2 , Aufried Lenferink 1 , Henk van Wolferen 3 , Cees Otto 1
Fig.1: Raman microscope objective integrated in the FIB-SEM vacuum chamber: (Left) Raman microscope objective integrated in the FIB-SEM (FEI NOVA
Nanolab 600) vacuum chamber. (Right) Raman microscope added onto the FIB-SEM vacuum chamber.
We present an integrated confocal Raman microscope
in a FIB - SEM. The integrated system
enables correlative chemical specific Raman,
and high resolution electron microscopic analysis
combined with FIB sample modification on
the same sample location. New opportunities in
sample analysis using correlative Raman-SEM,
and Raman – FIB – SEM are demonstrated on
different samples in materials and biological
sciences.
chamber, with the other components positioned
onto and outside the electron
microscope, as presented in figure 1.
The integration places no limitation on
the operation of either the Raman or the
FIB-SEM. Figure 1 (left) shows both the
Raman objective and integrated 3D XYZ
stage, used for sample scanning during
optical microscopy.
Introduction
The field of integrated Correlative Light
and Electron Microscopy (iCLEM) has
witnessed an enormous growth over
the last decade. Different optical microscopes
have been integrated in electron
microscopes, with the integration performed
by both commercial and scientific
organizations [1, 2]. In this article
a Raman microscope integrated with a
focused ion beam (FIB) – scanning electron
microscope (SEM) system is presented.
The commercial optical Raman
microscope, from HybriScan Technologies
B.V., is specifically designed for integration
in the SEM vacuum chamber.
It functions as an add-on module bringing
the optical objective into the vacuum
Fig. 2: Correlative high resolution SEM (A) and chemical specific Raman (B) analysis of multiple crystals,
and crystal polymorphisms. (C) Specific sample structures are identified with SEM, and the corresponding
Raman spectra are shown in (D, E, and F). Chemical specific Raman spectroscopy is used for
compound identification, showing: calcium sulfate (location 1, 2, 3, 4, 5), the calcium carbonate polymorphism
vaterite (location 6), the calcium carbonate polymorphism calcite (7), and multiple fluorescence
spectra from the photosynthetic bacteria M. aeruginosa [3].
34 • G.I.T. Imaging & Microscopy 2/2016