Experimental steering of electron microscopy studies using prior X-ray computed tomography

Area of Science:

• Biological imaging
• Correlative microscopy
• X-ray micro-computed tomography

Background:

• Electron microscopy (EM), including transmission electron microscopy (TEM) and scanning electron microscopy (SEM), offers high-resolution imaging but interrogates small, destructively prepared samples.
• Current EM workflows are time-consuming and lack broad contextual information, potentially leading to suboptimal data acquisition.

Purpose of the Study:

• To demonstrate how X-ray micro-computed tomography (microCT) can improve the efficiency and effectiveness of electron microscopy in biological research.
• To present a workflow integrating microCT with EM for enhanced sample selection, targeting, and data interpretation.

Main Methods:

• Utilizing X-ray micro-computed tomography (microCT) for initial sample assessment and 3D contextualization.
• Employing open-source software to integrate microCT data with subsequent electron microscopy (TEM/SEM) workflows.
• Developing strategies for site-specific targeting and guiding sample preparation based on microCT data.

Main Results:

• MicroCT enables precise selection of promising samples and specific regions of interest for EM.
• It provides multiscale correlative imaging, offering wider spatial context for EM investigations.
• MicroCT can guide EM sample preparation and 3D imaging schemes, and quantify preparation effects.

Conclusions:

• Integrating microCT prior to EM significantly enhances experimental efficiency and data quality in biological imaging.
• This correlative approach facilitates informed decision-making throughout the EM workflow, from sample selection to data analysis.
• The proposed workflow, leveraging open-source tools, offers a practical method for routine incorporation into biological research.