Categories
Trondheim

Viral Vector Core

The Viral Vector Core at Kavli Institute for Systems Neuroscience is a well-equipped facility with a rare expertise to make high quality viruses for research purposes. The Core is a non-profit facility that offers consultation, design and construction of a wide variety of viral vectors to augment scientific research.

Recombinant viruses are highly efficient gene delivery vehicles for introducing molecular tools to difficult-to-transfect primary cells, to create stable cell lines, to transduce tissues in vivo for elucidating underlying biological mechanisms and nowadays for gene therapy applications in clinics.

A combinatorial approach using viral vectors with the vast repertoire of transgenic animals available is a powerful strategy to target specific cells in complex organs such as mammalian brains. 

Adeno-associated virus, Moloney murine leukemia virus, Lenti virus and pseudotyped G-deleted rabies virus are all common transgene-delivery agents, each having distinct characteristics making them suitable for specific research questions.

Depending on the scientific question, one may need viral tools with specific features, demanding novel vector designs.

Recombinant viruses can be custom synthesized on a fee-for service basis, when the customer provides transgene construct for virus production. Vector core can assist in research by designing novel constructs and subsequent production of the viruses, please consult the core facility for the details. 

Categories
Trondheim

7Tesla magnetic-resonance imaging

Norwegian 7T MR Center is a National infrastructure for neuroscience research. The mission is to provide the Norwegian community of neuroscientists the very best tools for high resolution structure-function mapping of the brain.

7 Tesla Magnetic-Resonance (MR) will help to translate knowledge from model preparations in animals to the normal and pathological human brain. While studies of the human brain cannot currently match the spatial and temporal resolution obtained in animal preparations, a clinical 7T MR scanner will, with new technology for reduction of distortions and artefacts, allow us to image the human brain at a very high resolution. This radical improvement in resolution will pave the way for human studies of computation in subareas, layers and columns, which are much closer to the actual computational units of the brain than the global areas accessible with 3T scanners. The anticipated convergence of structural resolution in animal and human studies is likely to link concepts and hypotheses in the two research communities. This is for example critical for studies of entorhinal cortex, which is generally the first brain region to show degeneration in Alzheimer’s disease (AD). We envisage that high-resolution studies of structure, white matter connectivity and grey matter activity patterns on 7T MR scans can be used for subclinical diagnosis and monitoring of disease progression. 

The installed MR-system, a Siemens MAGNETOM Terra System, is equipped with:  

  • Dual Mode (Clinical and Research).  
  • 8 Channel RF-transmit chain in research mode  
  • 80/200 Gradient System.  
  • 32RX/1TX head coil (clinical mode)  
  • 32RX/8TX head coil (research mode only)  
  • 28RX/1TX knee coil (clinical mode)  
  • 13C and 31P loop coils  
  • Multinuclear option  
  • fMRI equipment  


Photo: Siemens Healthineers.  

Categories
Oslo

Mass spectrometry

The NORBRAIN mass spectrometry unit delivers high sensitivity for the quantitative analysis of small molecules, peptides, toxins and drugs. We provide expertise, services, education, and training to enhance biomedical research through mass spectrometry-based technologies.

Equipment and software

Thermo Scientific Q Exactive Hybrid Quadrupole-Orbitrap Mass Spectrometer

High resolution and accurate mass (HR/AM) analysis on the NORBRAIN Q‐Exactive mass spectrometer presents a unique configuration: the combination of the orbitrap mass analyzer with a quadrupole mass filter for precursor ion mass selection enables new quantitative methods based on HR/AM measurements, including targeted analysis in MS mode (single ion monitoring, SIM) and in MS/MS mode (parallel reaction monitoring, PRM).

The ability of the quadrupole to select a restricted m/z range allows overcoming the dynamic range limitations associated with trapping devices like the Orbitrap XL. Therefore high end quantitative and qualitative analysis in the area of in drug discovery, proteomics, environmental and food safety, clinical research and forensic toxicology can be offered. For example a proteomic discovery analysis on the Q-Exactive allows the detection of several thousand proteins in one run (180 min gradient).

Thermo Scientific TSQ Vantage Triple Stage Quadrupole Mass Spectrometer

Delivers high sensitivity for the quantitative analysis of small molecules, peptides, toxins and drugs. It is coupled online to a Dionex LC-system (Thermo-Fisher Scientific) and offers selected-reaction monitoring of peptide for protein quantification with an extended mass range of up to 4000 m/z.

Both complementary mass spectrometry systems (TSQ Vantage and Q-Exactive) therefore allow to develop high throughput biomarker analyses (TSQ Vantage) based on the suggested biomarker candidates from previous Q-Exactive MS analyses.

Software

  • Database search tools: Sequest (Proteome Discoverer 1.4)  for tandem mass spectra interpretation
  • MaxQuant/ SIEVE for relative quantification
  • In-house software tools (SAPA- and peptide mass tool) for protein peptide analyses

Services

We advise on epiproteomics, protein and lipid profile characterization as well as metabolomics, small molecule analysis, and analysis of post-translational modifications. We also offer services related to quantitative protein expression profiling (SILAC, N15 labelling, LFQ=label free quantification), and consult on experimental design and proteomic analysis options.

  • Protein analyses offered include in-gel or in-solution protease digestion, chromatographic separation and tandem mass spectrometric analysis of the generated peptides, and interpretation of MS/MS data using software like Sequest, Mascot, and MaxQuant.
  • The facility also assists especially in applications for the isolation (enrichment strategies like lectin chromatography), detection and characterization of post-translationally modified peptides (e.g. phosphorylation, N- and O-linked glycosylation, oxidation, ubiquitination, cysteine modifications, acetylation/ methylation (+ di- and tri-methylation) and O-GlcNAc modification). Sites of modification are verified by manual inspection of the data.

Please consult facility staff for projects which implements specialized approaches related to proteomic PTM analysis.


Photo: Gunnar F. Lothe, Institutt for medisinske basalfag, UiO.

Categories
Trondheim

Surgery facilities

Surgery facilities are available for users planning to perform experiments with instruments that are part of the NORBRAIN facility (tetode recordings, Neuropixels recordings, 2P microscope imaging, etc).  

Our surgery facilities are equipped with setup for anesthesia and surgery for intracranial delivery of substances or implantation of devices, in mice or rats. Necessary equipment for supportive therapy and postoperative care are also available, along with consumable materials and storage possibilities for medicines and test substances. 


Storage of medicines and test substances 

Ventilated cabinets are available for storage at room temperature, and fridges and freezers are available for products that need to be stored at cold temperatures.  

Instrument sterilization 

Surgical instruments must be sterilized before they can be used in surgeries. We have equipment for both dry heat sterilization and steam autoclaving. 

Specifications: 

Dry heat sterilization is done using a Termaks drying oven with adjustable temperatures and a display to control the temperature.  

For steam autoclaving we have a Tomy high-pressure steam sterilizer. 

Alternatively, glass bead sterilizers may be used to re-sterilize instruments.  

Anesthesia 

Animals must be anesthetized during surgeries. We have setup for gas anesthesia, which is easy to adjust to achieve suitable depth and a balanced anesthesia. 

Specifications: 

All surgery tables are set up with a vaporizer for either isoflurane or sevoflurane, with the possibility to use medical air and oxygen as the carrier gas. Medical air is delivered from a central system in the building. Oxygen is delivered from oxygen concentrators located in the surgery rooms. The gas anesthesia can be delivered to an induction chamber for induction of anesthesia, to a coaxial mask for maintenance of anesthesia, or to the stereotaxic mask for maintenance during the surgical procedure. The adjustment of the delivery pathway can easily be done by the user. To limit the exposure of the user, the surgery tables are down-drafted and we have an adjustable hood above the table. 

Stereotaxic setup 

The stereotaxic setup is especially designed to do find the correct area of the brain, with the use of precision instruments and coordinates based on anatomical landmarks. This gives the possibility to reach the exact area you are interested in. 

Specifications: 

The surgery tables are equipped with stereotaxic frames from Kopf Instruments, either a model 1430-B “U” frame for rats, or a model 900 “U” frame for mice. We have two main types of manipulators that can be used with the frame: 

  • Model 1760 Micro Manipulator (10 micron resolution): X, Z adjustment – Metric Vernier scale 80 mm travel, calibrated dial – 10 micron increments, 1.0 mm of travel per revolution. Y adjustment – Manual adjustment 100 mm each side of zero (A/P bar) 0.1 mm Vernier scale. 
  • Model 1460 Electrode Manipulator (100 micron resolution): X, Z adjustment – 80 mm travel calibrated 0.1 vernier scale (100 micron increments), 3.0 mm advance per revolution. Y adjustment – Manual adjustment 100 mm each side of zero (A/P bar) 0.1 mm Vernier scale 

The manipulators are used to locate the correct area, or as holders for drill, implants, syringes or pumps. 

Drill 

Specifications: 

Micro motor handpiece drills from Foredom Electric are available on the surgery tables. The drills are small and flexible, they have speed control and can be adjusted to either manual control or foot-pad control. The drills can be integrated with the manipulators on the stereotaxic setup. 

Systems for micro-injection 

When the surgery is done to deliver a small volume of a substance to a localized area in the brain, it is necessary to use special equipment to do this with high precision. 

Specifications: 

Several systems for delivery of substances in the microliter or nanoliter scale are available.  

  • Hamilton syringes have ultrafine needles and can deliver substance in the nanoliter scale. From Hamilton Company 
  • Nanoliter 2010 micro-injection system, which can be combined with a Micro 4 microsyringe pump controller, from World Precision Instruments 
  • Nanoject III programmable nanoliter injector, from Drummond Scientific Company 

Light source 

It is important to have sufficient light when doing surgeries. On the surgery tables we have lamps with flexible tubes so that the user can direct the light to the area where the light is needed. 

Specifications: 

The light source is LED connected to fiberoptic tubes. The user can adjust the direction of the light, and also the light strength, through a dimmer. 

Heat support 

Providing heat support during the surgery is important to maintain the body temperature of the rodents. 

Specifications:  

The surgery tables are equipped with an electric heat pad that is located on top of the platform where the animal is placed during surgeries. The heat pad is connected to a temperature controller with a display to monitor.  

Fluid support 

To prevent dehydration, fluids are given to the animals. Fluids are heated to body temperature.  

Specifications: 

A water bath with adjustable temperatures facilitates the preparation of body-temperature fluids which are given as fluid therapy to the animals during surgeries.  

Stereo microscope 

In order to get a good overview of the surgical field, the microscope gives a nice opportunity to see the area and make necessary adjustments. 

Specifications: 

We have stereo microscopes from Nikon and Zeiss, with adjustable focus and zoom. Some of the microscopes are connected to a camera that transfers the view to a screen, so that observers can follow the surgery. 

Ultrasound imaging 

One room has a high-frequency ultrasound imaging system that gives high-resolution images in space and time, of small anatomical structures. This gives the possibility to do image-guided procedures, for example. 

Specifications: 

The ultrasound scanner is a Vevo 1100 from Visualsonics, with a MS550S transducer (20-40 mHz and 16 mm penetration), an integrated rail system for positioning the animal and an image-guided injection system. A physiology monitor helps to follow the vital body functions during the procedure. The setup is connected to gas anesthesia. Available functionalities on the ultrasound scanner are M mode, B mode, colour doppler, pulse wave doppler, needle guide, and storage and post-processing of images and videos. 

Post-operative recovery chamber 

The surgical facilities are equipped with recovery chambers where animals can wake up from surgeries. The recovery chambers give the animals a warm and calm environment for the first hours after surgery. 

Specifications: 

The recovery chamber is a Maxi-Thermacage from Datesand. The chamber allows for precise temperature adjustment and control through a display. The temperature is evenly distributed inside the chamber. 

Other laboratory equipment 

The surgical facilities are also equipped with general laboratory tools such as shakers (for example MS3 basic, IKA), micro-sentrifuges (Mini Star, VWR), an ice cube machine (Porkka), and a water purification system (Direct-Q, Merck Milipore). 

Please note: Participating during animal experiments requires that the following conditions are met: 

  • People need to have completed the education and training in laboratory animal science as required by the national competent authority (The Norwegian Food Safety Authorities) equivalent to, at minimum, function (a) (persons who perform procedures on animals) according to the EU commission’s framework document for education and training. The local person responsible for animal care will need to evaluate the documents/certificates of the completed education and training. 
  • The experiments must be approved by the Food Safety Authorities, and the local animal welfare body must be oriented about the experiment in advance. The animal welfare body can set conditions for the experiment, based on local routines and guidelines 
  • People participating in the experiment must have been introduced to local routines regarding, for instance, health and safety, and use of the animal facility 
  • People must have been trained in the correct use of the equipment