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Femtosecond laser preparation of resin embedded samples for correlative microscopy workflows in life sciences

Bosch, Carles; Lindenau, Joerg; Pacureanu, Alexandra; Peddie, Christopher J; Majkut, Marta; Douglas, Andrew C; Carzaniga, Raffaella; ... Stegmann, Heiko; + view all (2023) Femtosecond laser preparation of resin embedded samples for correlative microscopy workflows in life sciences. Applied Physics Letters , 122 (14) , Article 143701. 10.1063/5.0142405. Green open access

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Abstract

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.

Type: Article
Title: Femtosecond laser preparation of resin embedded samples for correlative microscopy workflows in life sciences
Open access status: An open access version is available from UCL Discovery
DOI: 10.1063/5.0142405
Publisher version: https://doi.org/10.1063/5.0142405
Language: English
Additional information: © 2023 Author(s). All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http:// creativecommons.org/licenses/by/4.0/).
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Neuro, Physiology and Pharmacology
URI: https://discovery.ucl.ac.uk/id/eprint/10168271
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