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Article

Invadopodia Structure in 3D Environment Resolved by Near-Infrared Branding Protocol Combining Correlative Confocal and FIB-SEM Microscopy

1
Imaging Methods Core Facility at BIOCEV, Faculty of Science, Charles University, Průmyslová 595, 25242 Vestec u Prahy, Czech Republic
2
Department of Cell Biology, Charles University, Viničná 7, 12800 Prague, Czech Republic
3
Department of Physical Chemistry, Charles University, Hlavova 8, 12800 Prague, Czech Republic
4
Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (BIOCEV), Průmyslová 595, 25242 Vestec u Prahy, Czech Republic
*
Authors to whom correspondence should be addressed.
Academic Editor: Riccardo Alessandro
Int. J. Mol. Sci. 2021, 22(15), 7805; https://doi.org/10.3390/ijms22157805
Received: 27 May 2021 / Revised: 2 July 2021 / Accepted: 19 July 2021 / Published: 21 July 2021
Cancer cell invasion through tissue barriers is the intrinsic feature of metastasis, the most life-threatening aspect of cancer. Detailed observation and analysis of cancer cell behaviour in a 3D environment is essential for a full understanding of the mechanisms of cancer cell invasion. The inherent limits of optical microscopy resolution do not allow to for in-depth observation of intracellular structures, such as invadopodia of invading cancer cells. The required resolution can be achieved using electron microscopy techniques such as FIB-SEM. However, visualising cells in a 3D matrix using FIB-SEM is challenging due to difficulties with localisation of a specific cell deep within the resin block. We have developed a new protocol based on the near-infrared branding (NIRB) procedure that extends the pattern from the surface grid deep inside the resin. This 3D burned pattern allows for precise trimming followed by targeted 3D FIB-SEM. Here we present detailed 3D CLEM results combining confocal and FIB-SEM imaging of cancer cell invadopodia that extend deep into the collagen meshwork. View Full-Text
Keywords: FIB-SEM; CLEM; invadopodia; MT1-MMP; invasiveness FIB-SEM; CLEM; invadopodia; MT1-MMP; invasiveness
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MDPI and ACS Style

Dalecká, M.; Sabó, J.; Backová, L.; Rösel, D.; Brábek, J.; Benda, A.; Tolde, O. Invadopodia Structure in 3D Environment Resolved by Near-Infrared Branding Protocol Combining Correlative Confocal and FIB-SEM Microscopy. Int. J. Mol. Sci. 2021, 22, 7805. https://doi.org/10.3390/ijms22157805

AMA Style

Dalecká M, Sabó J, Backová L, Rösel D, Brábek J, Benda A, Tolde O. Invadopodia Structure in 3D Environment Resolved by Near-Infrared Branding Protocol Combining Correlative Confocal and FIB-SEM Microscopy. International Journal of Molecular Sciences. 2021; 22(15):7805. https://doi.org/10.3390/ijms22157805

Chicago/Turabian Style

Dalecká, Markéta, Ján Sabó, Lenka Backová, Daniel Rösel, Jan Brábek, Aleš Benda, and Ondřej Tolde. 2021. "Invadopodia Structure in 3D Environment Resolved by Near-Infrared Branding Protocol Combining Correlative Confocal and FIB-SEM Microscopy" International Journal of Molecular Sciences 22, no. 15: 7805. https://doi.org/10.3390/ijms22157805

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