Service overview
1. Sample preparation
Fixation, embedding (cryo, paraffin) and sectioning of samples.
Sample drop-off: Emmy De Blay (D007).
Cost (VUB):
- fixation, embedding + blocking: 2€/sample
- sectioning (charged slide): 2€/slide
- H&E, Oil Red O, PAS , beta-gal staining: 1,50€/slide
- self-use of micro/cryotome after training: 11€/hr
2. RNA/DNA in situ hybridization
RNA/Base/miRNA/DNAscope (Advanced Cell Diagnostics) represents a major advancement in RNA/DNA in situ hybridization employing a specific probe design that allows to amplify target-specific signals in (archival) tissue sections or cultured cells from any species with single-molecule detection sensitivity while simultaneously suppressing background noise. RNAscope targets longer (>300 bp) RNA including mRNA and lncRNA. BaseScope targets short (50-300 bp) RNA including highly homologous sequences, exon junctions, splice variants and point mutations. miRNAscope targets very short (17-50 bp) RNA including antisense oligonucleotides (ASOs), miRNA and siRNA. DNAscope is used to detect gene translocations and copy number variations. Depending on epitope survival, RNA/Base/miRNA/DNAscope can be combined with antibody-based protein staining.
Cost (VUB): upon request
3. Protein staining
Conventional antibody-based immunostaining or multiplex protein staining with DNA-barcoded antibody panels (Ultivue).
Cost (VUB):
- manual immunostaining (fluorescent/chromogenic): 16€/slide (primary antibody/ies not included)
- Ultivue: upon request
4. Image acquisition
Automated acquisition of brightfield and fluorescent (DAPI/GFP/RFP/Cy5/Cy7) images of slides and cultured cells at 4/10/20/40x (ThermoFisher Scientific EVOS M7000).
Automated high-throughput whole slide scanning in brightfield at 40x (Leica Aperio GT450) or fluorescence (6 channels) at 5/10/20/40x (Zeiss Axio Scan.Z1).
Cost (VUB):
- EVOS M7000: 11€/hr*
- Aperio GT450: 3€/slide* (trial: first 4 slides for free for new user)
- Axio Scan.Z1: 13€/slide (trial: first 4 slides for free for new user)
* self-use after training
5. Image processing, analysis & storage
Zeiss ZEN 3.5 blue image processing software.
Indica Labs HALO AI-assisted image analysis software.
Currently installed HALO modules:
>200 TB Network Attached Server storage with 10 Gigabit connection.
Cost (VUB):
- image processing/analysis computer: 11€/hr (self-use after training)
- data storage: 0,75€/GB/year
Testimonials
"Thanks to RNAscope performed at VUB, we were able to study in situ expression of the stem cell marker LGR5 in human colon, which would otherwise be impossible given the lack of reliable LGR5 antibodies."
A. Munoz, Instituto de Investigaciones Biomedicas "Alberto Sols", CSIC-UAM, Madrid
"We got a pretty positive experience with Yves' Team. The experiments were performed in a short time and we obtained very clear staining. RNAscope results were important to confirm our findings. Ultimately, Yves provided interesting advice and was very collaborative and available to answer our questions."
P. Jacquemin, de Duve Institute, Université catholique de Louvain, Brussels
“In collaboration with ACDBio and VUB, our laboratory designed a custom RNAscope probe against a target (Slc7a11/xCT) for which specific antibodies were lacking. The very first trial was successful and we moved on further to develop a fluorescent multiplex assay aiming at identifying the cellular expression.”
C. Nicaise, Namur Research Institute for Life Sciences, Université de Namur, Namur