Tuesday 1 June 2021

Correlative microCT and EM enabled by CyGEL mountant

In a comprehensive study to evaluate the properties of transplantable patient-derived gut organoids scientists at The Francis Crick Institute, Great Ormond Street Hospital and UCL used a suite of analyses, ranging from metabolic, biochemical and multi-lengthscale and multi-feature imaging techniques.  

In one iteration, they combined µCT and TEM/SBF-SEM/SEM montage. µCT was used to check the orientation of the cells on recellularized scaffolds prior to onward processing for electron microscopy.  To achieve this in a non-destructive manner that maintained the spatial integrity and orientation of the cellular objects and the scaffold, the fixed re-cellularized scaffolds were mounted in the thermo-reversible mounted CyGEL™. After CT scanning the sample was placed in ice to gently wash away the CyGEL and thereby recovered for onward EM processing.

This is the first demonstration of the use of CyGEL in µCT and in a correlative imaging technique. This transparency to x-rays, in addition to CyGEL's purpose-designed compatibility with fluorescence microscopy, give it great utility in multi-modal imaging techniques.

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Reference:

Meran, Laween, et al. "Engineering transplantable jejunal mucosal grafts using patient-derived organoids from children with intestinal failure." Nature Medicine 26.10 (2020): 1593-1601.

Links to related blogposts on correlative microscopy techniques:

CryoChem - from fluorescence to µCT to SBEM

ChromEMT - labeling chromatin

Simplified CLEM method - from in vivo imaging to FIB/SEM

Multi-scale tissue analysis: hybrid fluorescence-AFM


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