Histology and Imaging Facility

SEM image
SEM image of endothelial cells in phalanx pattern, lining a blood vessel
Histological analysis is a cornerstone technique to phenotype the transgenic animal models (mice and zebrafish). Soon after the transgenesis technologies were introduced in the CCB, an in house histology core was set up. This endeavour has now developed into a sophisticated state-of-the-art histology and imaging facility, allowing detailed and dynamic imaging of the neuro-vascular phenotypes of the animal (disease) models.

Histology

Managed by: David Nittner

The histology facility is equipped with the newest devices to perform automatic tissue embedding as well as paraffin, cryo-, vibratome-, and semi-thin sectioning.
We have also expertise in tissue preparation for scanning electron and transmission electron microscopy in collaboration with the Analytical Electron Microscopy Core Facility (Peter Baatsen, VIB-KULeuven).

zebrafish
Confocal microscopy image of tumor blood vessels in zebrafish brain


We routinely perform double and triple fluorescent immunostainings on all types of tissue sections (ie. muscle, brain, heart, tumor, ...) or cell cultures. Moreover, we have set up protocols for performing whole mount immunohistochemistry on small animals such as zebrafish and mouse embryos and retinas. In addition, techniques for in situ hybridization on tissue sections as well as on whole mount samples are set up, standardized and commonly used in the lab.   

Steering Committee

  •  Chris Marine
  •  Gabriele Bergers
  •  Anna Sablina
  •  David Nittner

 

Imaging

Managed by: Stefan Vinckier

SIM
Super resolution image (SIM) of mitochondria (in blue) within endothelial cells

The imaging facility is equipped with state-of-the-art hard- and software. Our facility is equipped with a motorized upright Leica DM5500 epifluorescence microscope, an inverted Leica SP8x confocal microscope with white light laser (470-670 nm), and an upright Leica SP8x multi-photon microscope (Holger Gerhardt) designed for intravital multi-photon imaging.

Furthermore, our facility is equipped with an upright Zeiss AxioPlan 2 epifluorescence microscope, a motorized inverted Leica DMI6000 B epifluorescence microscope and two inverted Zeiss confocal microscopes (C LSM 510 Meta NLO and LSM 780) especially designed for live cell imaging experiments (Peter Carmeliet). A laser capture microdissection system is available via access to a PALM MicroBeam 4 system  (Carl Zeiss) equipped with a temperature controled heating chamber, for laser microdissection of tissue sections or live cells under fluorescence (Colibri LEC illumination) or bright light illumination (subsidised by a Hercules Foundation grant, AKUL-46).  

The confocal microscopes enable us to perform FRAP, emission- and excitation fingerprinting, 3D-reconstruction and 3D-time-lapse imaging (4D-imaging) of living organisms and cells in the temperature-, humidity- and CO2-O2 controlled incubation chamber. We build up expertise in 3D-time-lapse imaging of several different cell types such as vascular endothelium cells, pericytes, neurons, as well as electroporated brain slices and zebra fish in normoxic and hypoxic conditions.

Morphometric (stereological) analysis occurs through semi-automatic macros written with the latest Leica MetaMorph software packages, enabling us to analyze several dozens of parameters.

We actively collaborate with other researchers and core facilities to perform state-of-the-art imaging, such as superresolution imaging (dSTORM/PALM, Johan Hofkens, KULeuven); second harmonic generation and Airyscan imaging (Pieter Vanden Berghe, Cell Imaging Core, KULeuven); Spinning disk, SIM, TEM, SEM and FIB-SEM imaging  in close collaboration with the VIB Bio Imaging Core (Sebastian Munck, Chris Guerin, Saskia Lippens, Natalia Gunko).  

 

For an overview on the expertise of the different microscopy core facilities at the Biomedical Sciences Group (KULeuven) and the VIB Bio Imaging Core we refer to:

http://gbiomed.kuleuven.be/english/corefacilities/microscopy
http://bio-imaging-core.be/

Our facility is also a member of the European Light Microscopy Initiative.

References:

  • The Cancer Cell Oxygen Sensor PHD2 Promotes Metastasis via Activation of Cancer-Associated Fibroblasts. Kuchnio A, Moens S, Bruning U et al. & Carmeliet P. Cell Rep. 2015 Aug 11;12(6):992-1005
  • Fatty acid carbon is essential for dNTP synthesis in endothelial cells. Schoors S, Bruning U, Missiaen R, Queiroz KC, et al. & Carmeliet P. Nature. 2015 Apr 9;520(7546):192-7.
  • Tumor vessel normalization by chloroquine independent of autophagy. Maes H, Kuchnio A, Peric A, et al. & Carmeliet P. Cancer Cell. 2014 Aug 11;26(2):190-206.
  • Partial and transient reduction of glycolysis by PFKFB3 blockade reduces pathological angiogenesis. Schoors S, De Bock K, Cantelmo AR et al. & Carmeliet P. Cell Metab. 2014 Jan 7;19(1):37-48
  • Role of PFKFB3-driven glycolysis in vessel sprouting. De Bock K, Georgiadou M, Schoors S et al. & Carmeliet P. Cell. 2013 Aug 1;154(3):651-63. Mazzone M et al. (2009) Heterozygous Deficiency of PHD2 Restores Tumor Oxygenation and Inhibits Metastasis via Endothelial Normalization. Mazzone M et al. (2009). Cell 136: 839-851