Microscopic imaging and technology

The research within our LUMC departments is conducted within departmental research programmes. The research programme below is embedded within the department of Chemical Cell Biology / Molecular Cell Biology.

Aim and focus

We have two ongoing lines of biological research. One line is focused on Virus Replication (Montse Barcena) and the other on Bionanopatterning (Thom Sharp). For all our research we use our state-of-the-art electron microscopy infrastructure, image processing and visualization techniques and correlative light and electron microscopy. In many cases for specific research projects we further develop or adapt methods and techniques.
We have two lines of technology-related research. One is on Correlative Light and Electron Microscopy (Roman Koning) and the other on Advanced Image Acquisition and Processing (Frank Faas).  Aim of the technology-related research is to develop and/or adapt microscopic imaging methods, instrumentation and fluorescent labelling technologies to study the molecular composition of tissues, cells and sub-cellular compartments. The programme involves both light and electron microscopy and supports projects within the realm of both cell- and structural biology.
In addition to the technology-related research, we provide microscopy technology-related services in the framework of a LUMC Technical Focus Area Microscopy (www.kosterlab.nl). The TFA Microscopy is composed of two main imaging technologies: Electron Microscopy and Light Microscopy. The TFA is closely affiliated with the high-resolution cryo-electron microscopy facility (NeCEN) at the Institute of Biology Leiden at Leiden University. Heading the TFA is Lennard Voortman, who is also coordinating the Light Microscopy group. The Electron Microscopy part of the TFA is coordinated by Roman Koning.
Examples of light microscopy developments are two-photon or confocal laser scanning microscopy to study molecular dynamics in living cells using, time-lapse fluorescence imaging of living cells, two-photon intravital microscopy of small laboratory animals, digital fluorescence workstations to perform 48 colour FISH karyotyping in case of malignancies and pre/postnatal abnormalities. For high-resolution (nm-scale) ultrastructural electron-microscopy, the core electron microscopic imaging tools include (cryo) transmission electron microscopy (TEM) with energy-filtering, (cryo) 3D scanning electron microscopy (3D SEM) and electron tomography (3D imaging of cellular organelles and macromolecules). Ongoing technical developments aim at improving specimen preparation and labelling techniques and at advancing further automation of methodologies. A key development of the Research Programme is aimed at correlative light-electron microscopy, allowing the combination of high-resolution imaging with electron microscopy with live-cell dynamic imaging with fluorescence light microscopy.
In addition the group develops cytochemical staining procedures and dedicated software for cell and structural analysis. Applications of microscopic imaging and technology range from fundamental cell biological studies, imaging the cellular ultrastructure and/or dynamics with electron microscopy or live cell microscopy, to diagnostic applications of multicolour FISH and reported technology in case of oncology, genetics and infectious diseases. 

Position in international context

The group has an international reputation due to its pioneering role in the development of multicolour FISH techniques, fluorescence microscopy and microscopy automation, 3D electron microscopy, high resolution cryo-electron tomography and correlative light and electron microscopy. Consequently, the group has organised many EMBO workshops (on FISH technology), Marie Curie courses (on genomic profiling), EMBO courses (on electron tomography) and takes part in several European consortia.

Content / highlights / achievements

  • Discovery of a new role of the NFATc2 gene in a Ewing sarcoma variant (with Department of Pathology) 
  • Development of lab-on-a-chip systems for point of care (POC) diagnosis 
  • An intravital multi-photon microscopy system has been made available for investigation of small laboratory animals and is currently used by several LUMC groups. 
  • Construction of a light-microscope system enabling fluorescence imaging of cryofixed biological specimens prior to observation with cryo-electron microscopy. 
  • Better insight into the replication of +RNA viruses by live cell imaging, CLEM and 3D-SEM (collaboration with the Department of Medical Microbiology). 
  • Discovery on how Complement is activated by the immune system on a molecular scale using high resolution cryo electron microscopy (collaboration with Utrecht University). 
  • Better understanding of antibody-binding using high resolution electron tomography. 
  • Better insight in the structure of the glycocalyx in the kidney (collaboration with the department of Nephrology) by the application of large-scale electron microscopy imaging. 

Future themes

Method development to increase the efficiency and quality of advanced electron microscopy techniques by further automation of data acquisition and visualization. Techniques to combine fluorescence light microscopy with high-resolution cryo electron tomography are in development, focusing on identifying and imaging molecular assemblies in situ without chemical or gold labeling pre-treatments. Stimulate more use ultra-high resolution light microscopy (PALM, STED) for application within the LUMC.

Cohesion within LUMC

The light and electron microscopy facilities and infrastructure function as a TFA and are extensively used for collaborative research and development projects within the LUMC. Important themes of collaborative research are genetics, pathology, oncology, haematology and infectious diseases.