Department Imaging Physics
Principal investigator Jacob Hoogenboom
E-mail address j.p.hoogenboom@tudelft.nl
Website https://www.hoogenboomlab.org/
3D “Google Maps” for biology
We want to achieve faster and automated imaging so that an entire tumour or an organ like the brain can be imaged in a “Google Maps” like fashion.
We extract information out of the data with computational methods for label-free prediction of fluorescence and automated segmentation. We focus on correlative light and electron microscopy (CLEM) as well as large volume EM using multibeam SEM.
Techniques
- CLEM
- SEM
- Multibeam SEM
- Array tomography
Further reading
Kievits, Arent J., Duinkerken, B. H. Peter, Lane, Ryan, de Heus, Cecilia, van Beijeren Bergen en Henegouwen, Daan, Höppener, Tibbe, Wolters, Anouk H. G., Liv, Nalan, Giepmans, Ben N. G. and Hoogenboom, Jacob P.. “FAST-EM array tomography: a workflow for multibeam volume electron microscopy” Methods in Microscopy, vol. 1, no. 1, 2024, pp. 49-64. https://doi.org/10.1515/mim-2024-0005
Localization for cryo-FIB-SEM
With Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) proteins of interest can be carved out of a cell for high resolution cryo-TEM. However, finding the protein of interest inside the cell is like finding a needle in a haystack. We have developed 3D fluorescence localization to pin-point the protein for focused ion beam milling in cryogenic conditions.
Techniques
- FIB-SEM
- Cryo-TEM
- Fluorescence microscopy
Further reading
Daan B Boltje, Jacob P Hoogenboom, Arjen J Jakobi, Grant J Jensen, Caspar TH Jonker, Max J Kaag, Abraham J Koster, Mart GF Last, Cecilia de Agrela Pinto, Jürgen M Plitzko, Stefan Raunser, Sebastian Tacke, Zhexin Wang, Ernest B van der Wee, Roger Wepf, Sander den Hoedt (2022) A cryogenic, coincident fluorescence, electron, and ion beam microscope eLife 11:e82891 https://doi.org/10.7554/eLife.82891
Superresolution E-beam calibration
Together with our colleagues at Erasmus MC, we have developed a technique to print biomolecules at very small (sub-100 nm) length scales. We use these biomolecular patterns for calibration and performance testing of super-resolution light microscopes. Projects can focus on improving the resolution and extending its application range.
Techniques
- Superresolution microscopy