1. Superovulation for embryo collection
In order to be able to collect embryos at the 2 cell-stage for freezing or direct embryo transfer into axenic recipients, the following protocol for superovulation of the females, mating and dissection must be followed strictly for a successful recovery of the embryos (CMF_Superovulation).
2. Quarantine and hygiene restrictions for CMF access
The CMF underlies a strict hygiene protocol in order to maintain the axenic and gnotobiotic character of the individual rooms.
Access to the CMF is only authorized to a small group of researchers after introduction and training by the head of the facility. We will discuss together if access is necessary for your experiment or if our technicians can perform the required manipulations.
Entry into the CMF is only possible after a 14-day quarantine period in which no contact to other rodents (SPF facility) has happened. This includes also presence in rooms that contain non-gnotobiotic mice. Personnel with rodents as pets are forbidden to enter the CMF at any times.
3. Preparation and drop of experimental material to enter isolators (autoclavable)
The material required for your experiment (such as ear clips, tubes, capillaries for bleeding, bottles), which needs to be introduced into your experimental isolator, should be autoclavable and it MUST enter the isolator via a drum. Please, prepare the material inside a glass bottle with lid or a metal box and proceed to autoclave the material in your lab prior to bringing it to the CMF. Label all material with the experimental isolator number, date and your name. The autoclaved material needs to be brought to the back of the facility, deposited in the allocated place and an email explaining your import needs to be written to (email@example.com).
Import time is 5-10 days.
4. Preparation and drop of experimental material to enter isolators (non-autoclavable)
Products for the treatment of your mice that cannot be autoclaved (certain heat-unstable chemicals, bacteria for colonizations etc.) needs to prepared in a sterile way following the protocol “CMF_Preparation of non-autoclavable material” to make sure no unwanted microbes enter the facility and contaminate the experimental isolator. Briefly, all used material needs to be cleaned and autoclaved before preparing your import kit. Preparation of the import kit needs to take place in a laminar flow hood using a sterile gown and sterile surgical gloves. The final import kit needs a double container system of which only the outside of the outer container is not sterile. The inner container should consist of a glass bottle with a tightly closed lid. This inner bottle will be sterilized by a 4% peracetic acid solution sprayed directly into the port of the isolator. Decontamination of the inner bottle by peracetic acid takes 45 minutes. Only after this time, you are allowed to open the inside door of the isolator port.
Please, contact us for training and instructions. Each new type of import kit needs to be verified by one of the heads of the facility before first use.
5. How to fill in your experimental sheet
Please, find an example (CMF_Example experimental sheet) and a blank experimental sheet (CMF_Experimental sheet) in the download section under contacts.