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

Histology
The histology facility is equipped with the newest device to perform automatic tissue embedding as well as paraffin, cryo-, vibratome and semi-thin sections. We routinely perform double and triple fluorescent immunostainings on all type of tissue sections (ie. muscle, brain, heart, tumor…). Moreover, we have set up protocols for performing whole mount immunohistochemistry on small animals such as zebrafish, drosophila, Xenopus and mouse embryos. In addition, we have scaled down and miniaturized procedures to immunostain micro-dissected tissues such as, for instance, spinal cords of Xenopus tadpoles. In addition, techniques for in situ hybridization on tissue sections as well as on whole mount zebrafish, tadpoles and mouse embryos are set up, standardized and commonly used in the lab.

Imaging
The imaging facility is equipped with state-of-the-art hard- and software. Two Zeiss LUMAR fluorescence stereomicroscopes offer help for screening of zebrafish, tadpoles and Drosophila fruitflies and other fluorescent objects.  Furthermore, our facility is equipped with a Leica DM RBE research-microscope and three motorized Zeiss research-microscopes: an AxioPlan 2 imaging, an AxioVert 200M and an Imager Z1.  All microscopes are equipped with a computer and a Zeiss AxioCam HRc, MRc5 or MRm digital camera with a resolution up to 12M pixels for live imaging and acquisition.  Morphometric (stereological) analysis occurs through semi-automatic macros written with the latest Zeiss AxioVision and KS300-software, enabling us to analyze several dozens of parameters and control the microscope at once. A Zeiss CLSM510 confocal microscope is used for regular scanning, 3D-reconstruction of the specimen, time-lapse, FRAP-experiments and colocalization studies.  Excitation wavelengths ranges from 488 nm (Argon laser) over 546 nm (HeNe-laser) to 633 nm (IR, HeNe-laser). Our latest achievement is the Zeiss CLSM510 META NLO, equipped with the Coherent Chameleon laser for multiphoton microscopy, Argon- & two HeNe-lasers for normal confocal microscopy.  This microscope enables emission- and excitation fingerprinting, FRET, FRAP, FLIM, 3D-reconstruction and 3D- time-lapse video imaging (4D-imaging) of living organisms and cells in the temperature & CO2-controlled incubation chamber.

Microscopy reservations

References

Stefan Vinckier
Senior Scientist

Ann Manderveld
Senior Labtechnician

An Carton

Naima Dai

Kristel Peeters

Bjorn Van Wetswinkel