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CHESS imaging has previously been used to investigate insect larval development in a non-cryobiological context, and technological developments in the MR field have recently seen in vivo spatial resolution in non-frozen samples reach the size range relevant for entomological and subcellular investigations.
The present study combined these two fields of investigation by demonstrating the feasibility of high-resolution MR imaging of insect larvae in a cryobiological context.
Results
In vivo MR images were acquired from autumn-collected larvae at temperatures between 0°C and about −70°C and at spatial resolutions down to 27 µm. These images revealed three-dimensional (3D) larval anatomy at a level of detail currently not in reach of other in vivo techniques. Furthermore, they allowed visualization of the 3D distribution of the remaining liquid water and of the endogenous cryoprotectants at subzero temperatures, and temperature-weighted images of these distributions could be derived. Finally, individual fat body cells and their nuclei could be identified in intact frozen Eurosta larvae.
Conclusions
These findings suggest that high resolution MR techniques provide for interesting methodological options in comparative cryobiological investigations, especially in vivo.
