The Francis Crick Institute
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Femtosecond laser preparation of resin embedded samples for correlative microscopy workflows in life sciences

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journal contribution
posted on 2023-04-11, 11:27 authored by Carles Bosch, Joerg Lindenau, Alexandra Pacureanu, Christopher J Peddie, Marta Majkut, Andrew C Douglas, Raffaella Carzaniga, Alexander Rack, Lucy Collinson, Andreas T Schaefer, Heiko Stegmann
Correlative multimodal imaging is a useful approach to investigate complex structural relations in life sciences across multiple scales. For these experiments, sample preparation workflows that are compatible with multiple imaging techniques must be established. In one such implementation, a fluorescently labeled region of interest in a biological soft tissue sample can be imaged with light microscopy before staining the specimen with heavy metals, enabling follow-up higher resolution structural imaging at the targeted location, bringing context where it is required. Alternatively, or in addition to fluorescence imaging, other microscopy methods, such as synchrotron x-ray computed tomography with propagation-based phase contrast or serial blockface scanning electron microscopy, might also be applied. When combining imaging techniques across scales, it is common that a volumetric region of interest (ROI) needs to be carved from the total sample volume before high resolution imaging with a subsequent technique can be performed. In these situations, the overall success of the correlative workflow depends on the precise targeting of the ROI and the trimming of the sample down to a suitable dimension and geometry for downstream imaging. Here, we showcase the utility of a femtosecond laser (fs laser) device to prepare microscopic samples (1) of an optimized geometry for synchrotron x-ray tomography as well as (2) for volume electron microscopy applications and compatible with correlative multimodal imaging workflows that link both imaging modalities.


Crick (Grant ID: CC2036, Grant title: Schaefer CC2036) Crick (Grant ID: CC1076, Grant title: STP Electron Microscopy) Engineering and Physical Sciences Research Council (Grant ID: EP/W024292/1, Grant title: EPSRC EP/W024292/1)