High Throughput Microscopy Dutch Flagship Node
The Dutch High Throughput Microscopy flagship Node brings together the three leading screening centers in the Netherlands that have extensive experience with cell-based screening coupled to high content imaging and advanced microscopy. The integration of these three sites in one NL-HTM center creates a world leading expertise center in high-throughput microscopy for imaging based screening. Each center offers specific strength and expertise. Each of these sites has developed a high-end infrastructure covering a broad range of screening and imaging technologies. Leiden Cell Observatory is in particular experienced in high-throughput high-end microscopy using automated live cell time-lapse confocal microscopy in combination with focused RNAi and compound screening strategies. The CSC is specialized in automated high-throughput imaging based screens using low resolution high-throughput endpoint imaging. The NKI Screening Center is highly experienced and at the forefront in whole genome RNAi screening using genome-wide shRNA as well as siRNA libraries and the NKI Advanced Microscopy Center integrates high-end microscopy approaches into medium throughput automated microscopy settings.
Specialties and expertise of the Node
The Dutch HTM flagship Node is specialized in the development and application of cell based screening assays in 2D and 3D with image-based assays and readouts. Screening platforms compromise small molecule collections, functional genomic screening collections including CRISPR, shRNA and siRNA. Special interest and expertise lies in the use of advanced fluorescence based reporter cell lines to monitor live signaling dynamics. For automated image analysis, screen analysis and data integration we have developed relevant image analysis and bioinformatics pipelines using different open sources software for image quantification. Another area of expertise is the use of quantitative data for computational modelling to obtain a mechanistic understanding of observed phenotypes. We also have extensive experience in combining HCS with functional imaging assays, employing e.g. FRET sensors for readout of second messenger cascades, receptor activation and receptor internalization. New fast FLIM approaches have been implemented on our automated microscopes as well.
ISIDORe is a Horizon Europe funded project that brings together 154 partners from 32 countries around the world, and is designed to effectively support research on infectious diseases and increase preparedness for pandemic.
Additional services offered by the Node
- Technical assistance to run instrument
- Assistance with experimental design (e.g. experimental set up, choice of technique)
- Training in infrastructure use
- Probe preparation
- Animal preparation
- Wet lab space
- Server Space
- Data processing and analysis
- Training workstations
- Training seminar room
- Biobanking, biological material storage and processing
- Cell culture facilities - Safety level 1 and 2
Special features of instruments at the NL-HTM node: 1) we have both high-end confocal microscopes all equipped for automated screening and low-end HTM imagers; 2) all microscopes set-ups are equipped for live cell imaging; 3) automated robotics dedicated to perform siRNA and compound screenings are associated with the HTM imagers. We have confocal microscopes capable of adding agonists during acquisition, and we provide fast FLIM readout for FRET detection.
Dr. Roderick L. Beijersbergen
Asst. Prof. Divison of Molecular Carcinogenesis and Head of NKI Robotics and Screening Center
Dr. David A.Egan
Lab Manager, Department of Cell Biology UMC
Prof. Dr. K. Jalink
Head of Biophysics and advanced imaging group
+31 20 512 1933
Prof. Dr. Bob van de Water
Head of division of Toxicology