From molecules and cells...

From molecules and cells...

In research theme 1 we want to understand how molecular structural aberrations lead to cellular abnormalities. Within this research theme we will extend technology developed in Cyttron I for applications in pathology. At the same time we work on expanding the range of high resolution imaging by cryo-electron microscopy.

WP1.1 Towards near atomic resolution for single-particle cryo-electron microscopy
Work package 1.1
WP1.1 Towards near atomic resolution for single-particle cryo-electron microscopy

X-ray crystallography is the commonly used method for unravelling the 3D structures of proteins. Only about 8% of proteins crystallise and only 3% produce crystals big enough to solve with this technology. We want to optimise cryo-transmission electron microscopy to (i) solve the atomic structures of proteins that only produce nano-crystals and (ii) determine intermediate resolution structures (0.4 to 0.3 nm resolution) of protein complexes that fail to crystallise. We focus on electron detection, microscope data generation and data analysis.

Figure: Inside of the NeCEN Titan Krios cryo-transmission electron microscope

Work package leader(s): 
Frank de Jong, Dr.
Jan-Pieter Abrahams, Prof. dr.
Raymond Wagner, Dr.
Contact: 
Raymond Wagner, Dr.
WP1.2 FIB-SEM and correlative electron microscopy
Work package 1.2
WP1.2 FIB-SEM and correlative electron microscopy

In order to understand diseases and diagnoses, it is important to find specific cells in an overview image and then zoom in to high magnifications for detailed information. This can be achieved by correlative microscopy. We will work on two types of correlative microscopy: 1. Focused-Ion-Beam-Scanning-Electron Microscopy (FIB-SEM) and 2. the combination of Transmission Electron Microscopy (TEM) with fluorescence microscopy. We focus on further developing and applying these types of correlative microscopy in the vascular field.

Figure: Typical microscopy images of combined transmission electron microscopy (TEM) and fluorescence microscopy (FM).

 

Work package leader(s): 
Bram Koster, Prof. dr.
Jan Andries Post, Dr.
Raymond Wagner, Dr.
WP1.3 New tools for integrated, high-throughput, high-resolution imaging
Work package 1.3
WP1.3 New tools for integrated, high-throughput, high-resolution imaging

In this work package we will identify genes and study processes associated with inflammation cancer progression and cardiovascular diseases. This information is anticipated to lead to more accurate prognosis, diagnosis and/or staging of the diseases. We will use and develop various microscopic techniques, including high-resolution microscopy (STED micoscopy), high throughput high content microscopy (automated confocal microscopy) and intravital live cell imaging in the intact organisms   (multiphoton microscopy). This will be combined with high throughput screening devices for studying diseases in zebrafish embryos and RNAi screens for identifying disease-related genes. We will also investigate responses to disease-related chemical gradients. Integrating the different technologies is expected to lead to valuable new scientific insights with great potential for novel drug target identification.

Figure: The spread of cancer cells (red) in a fluorescent zebrafish embryo (green). Top: whole embryo, bottom: close-up showing metastasis in detail. Image from VPS Ghotra, S He, H de Bont, W van der Ent, HP Spaink, B van de Water, BE Snaar-Jagalska, EHJ Danen. Automated whole animal bio-imaging assay for human cancer dissemination. PloS One, in press 2012.

Work package leader(s): 
Bob van de Water, Prof. dr.
Hans Tanke, Prof. dr.
Herman Spaink, Prof. dr.
Thomas Schmidt, Prof. dr.
11/12/2014