Routine technologies

Microscopystereomicroscoop

Epifluorescence-Zeiss

The Zeiss Axiovert 200M is a fully automated invert microscope, suitable for light, phase-contrast and epifluorescence visualization. The long-distance lenses (4x, 10x, 20x, and 40x (with and without immersion oil) support the use of both culture flask/plates as slides. A set of 4 filters is installed allowing visualization of the most current fluorophores Image acquisition is supported by Axiovision 4.1 software.

Light microscopy Leica DM 705

The Leica DM 705 is a standard light microscope for tissue slides. It is equipped with a 4x, 10x, 20x, 40x and 100x (immersion oil) lenses.

Leica phase contrast

The Leica DMI 3000B is an inverted phase-contrast microscope with long-distance lenses (x5, x10, x20 and x40), suitable for culture flasks and plates.

Fluorescence strereomicroscope Leica M205 FA

The stereomicroscope is equipped with optics magnifying 7.8x – 160x and provides the possibility to view the material with visible light or using fluorescence (blue, red and green). All Leica microscopes are supported by the LAS4.1 software packet.

Q-PCRRoche-LightCycler

Real-time quantitative PCR (qPCR) is the standard method to quantitatively measure the expression levels of selected genes (genes of interest) in a fast, sensitive, selective and accurate way. Regardless of its ease of use, a successful qPCR experiment depends on smart experimental design, a proper choice of primers and reference genes, inclusion of quality controls and reliable data analysis software. LECR is equipped with a LightCycler 480 II instrument (384-well plate format, Roche) and performs its subsequent data analysis using qbase+ software (Biogazelle).

 

ReferencesVergauwen, G., et al., Confounding factors of ultrafiltration and protein analysis in extracellular vesicle research. Scientific Reports, 7(1), 12. en Van Deun, J., et al., The impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling. J. Extracell. Vesicles., 3, 2014. (I.F. 2014 = NA) (ranking: NA) (new journal) (citations: >185)

Biofluid analysisHematology-XP300

The XP-300 is a hematology analyzer and includes counting of red blood cells, white blood cells and platelets.

 

 

Protein analysisProtein analysis

The BioDrop (Isogen Life Science) is a UV/Vis spectrophotometer for the measurement of DNA, RNA, oligonucleotides and protein samples. The Qubit 3.0 (Invitrogen) is a benchtop fluorometer for measurement of DNA, RNA, oligonucleotide and protein quantity or quality. Because this technique makes use of fluorescence it is more sensitive than UV absorbance-based quantification. The lab is fully equipped to perform SDS-PAGE and Western blot analysis. In this way proteins can be separated based on their molecular weight and further analysed by immunostaining. Imaging of the blot is performed by the Proxima 2850 Imager (Isogen Life Science). The Proxima can also be used for imaging of ethidium bromide-stained DNA agarose gels, coomassie stained gels, SYBR Green stained gels, Petri dishes, etc.

 

Reference: Vergauwen, G., et al., Confounding factors of ultrafiltration and protein analysis in extracellular vesicle research. Scientific Reports, (2017), 7(1), 12.

 

Cell and tissue culture

LECR is fully equipped to perform cell and tissue culture activities including, but not limited to, adherent and suspension cells, 2D and 3D culture, matrix (type I collagen, Matrigel) culture, primary cells (fibroblast, mesothelial cells,...), established cancer cell lines, patient-derived cancer cells (PDC), hTERT-immortalized fibroblast, fluorescent and luminescent cancer models. The lab has a biobank of >8000 cryovials in collaboration with Bimetra. The lab maintains 7 CO2 incubators, one CO2 incubated rotary shaker and 8 LAF. The lab maintains the following technologies: Countess cell counter, Gentlemacs, Amaxa nucleofection, Eppendorf centrifuges, ...