RT Journal Article
SR Electronic
T1 Femtosecond laser preparation of resin embedded samples for correlative microscopy workflows in life sciences
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2023.01.10.523473
DO 10.1101/2023.01.10.523473
A1 C Bosch
A1 J Lindenau
A1 A Pacureanu
A1 CJ Peddie
A1 M Majkut
A1 AC Douglas
A1 R Carzaniga
A1 A Rack
A1 L Collinson
A1 AT Schaefer
A1 H Stegmann
YR 2023
UL http://biorxiv.org/content/early/2023/01/12/2023.01.10.523473.abstract
AB 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-labelled 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 (SXRT) or serial blockface scanning electron microscopy (SBF-SEM) 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 novel femtosecond laser device to prepare microscopic samples (1) of an optimised geometry for synchrotron X-ray microscopy as well as (2) for subsequent volume electron microscopy applications, embedded in a wider correlative multimodal imaging workflow (Fig. 1).Competing Interest StatementJL, ACD and HS are employees of Carl Zeiss Microscopy GmbH, the manufacturer of the femtosecond laser and of the Zeiss Crossbeam FIB-SEM employed and evaluated in this study. The authors have no further relevant interests to disclose.