Technology
Zebrafish CRISPR Knock-Out (KO) Service
Promotor, regulatory elements and exon deletions can be ordered at an extra cost.
Package | Target Validation | Target Validation | Injection | Injection | Full Package | Full Package |
|---|---|---|---|---|---|---|
sgRNA & | + | + | + | + | + | + |
In vivo sgRNA | + | + | + | + | + | |
Embryos injection | + | + | + | + | ||
Line propagation | + | + | ||||
Morphometric | + | + | + | |||
Cost (SEK) | 1 500 | 7 500 | 12 000 | 16 000 | 22 000 | 27 000 |
Zebrafish CRISPR Knock-In Service
Standard service includes insertion of the GFP protein under the expression of the endogenous gene promoter. Other methods can be applied at the customized pricing model.
Package | Target Validation | Target Validation | Injection | Full Package |
|---|---|---|---|---|
sgRNA & | + | + | + | + |
In vivo sgRNA | + | + | + | |
Embryos injection | + | + | ||
Line propagation | + | |||
Cost (SEK) | 1 500 | 7 500 | 18 000 | 26 000 |
Live imaging with high throughput systems
1. Live Imaging with high throughput automated VAST system (Union Biometrica)
Specification
Embryo and larvae 2-6 days post fertilization, 96-well plate based, bright field and fluorescent (green and red) imaging, magnification: 4X, 5X, 10X, (20X), automated positioning of the fish, image analysis option
Price: Flat rate of 1000 sek/run + fish handling (650 sek/h)
Example images acquired with VAST system. For all images, basic bright field reference images are taken. Example of lateral images of 2- and 5-days post fertilization (dpf) zebrafish embryos with automated area recognition (left panel). Fluorescent image of the double transgenic zebrafish trunk at 5 dpf.
2. Live Imaging with high content Acquifer Imagin Machine (Burker)
Specification
Embryo and larvae 2-5 days post fertilization, 96-well plate based, bright field and fluorescent (green and red) imaging, magnification: 2X, 4X, 10X, temperature control, manual fish positioning, image analysis option.
Price: Flat rate of 1000 sek/run + fish handling (650 sek/h)
Application: Temperature sensitive screens (e.g. heart rate screens)
Toxicity tests
Fish embryo acute toxicity (FET)
Specification
Test based on the adapted Test Guideline 236. This test is designed to determine acute toxicity of chemicals on embryonic stages of fish. Recommended by the EU grant committee as an intermediate step between in vitro and in vivo mouse studies.
Cardiotoxicity test
Specification
This test is designed to determine cardiac toxicity of chemicals on the embryonic fish heart
Automated imaging of the zebrafish embryonic heart combined with image analysis. Control embryo (left) and compound treated embryo (right) at 5 days post fertilization. Heart area is automatically detected and marked in red (upper panel), heart rate and contraction intensity are measured based on the video recording of the fish (lower panel).
Cancer modelling
STEP 1 – Evaluation of the human cancer cell line xenografts in the zebrafish embryo
Specification
An assay designed to evaluate cancer cell line compatibility with zebrafish model, including cell size, behaviour, survival, engraftment side and embryo toxicity.
Requirement
- Cell survival was tested in vivo at 33°C
- Cells are fluorescently labelled
- Cells are provided in 0.25-0.5x106 cells/ul concentration
STEP 2 – Tumour growth & metastasis assay
Specification
Customized experimental set up based on the results of the evaluation assay, injections at 1 or 2 dpf into the blood flow, brain cavity or PVC space. Imaging of the xenotransplanted fish at 2 time points. Possibility of combining xenotransplantation procedure with the drug treatment.
Cancer modelling in the zebrafish. Dorsal view of a zebrafish casper embryo injected in the brain ventricle with fluorescently labelled human medulloblastoma cells (upper panel). Tail of a zebrafish embryo with breast cancer xenografted cells (green) extravasating from the blood vessels and interacting with zebrafish macrophages (red) (lower panel).
Example of an experimental setup for tumour growth and metastasis assay.