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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