Biotechnology lab

General

You can find more info about our different services by clicking on the tab "Procedures". These services can also be ordered by external clients provided there are capacities.

Here you can find our forms.

Procedures

PN / Cytoplasm / Perivittelin microinjection

Mouse (positive foanders not guaranteed).
Injection of min 200 C57Bl/6NCRL Oocytes, other strains on demand
Oviducttransfer in 10 recipients
Rat (positive foanders not guaranteed).
Injection of min 200 Spraque Dawley oocytes, other strains on demand.
Ampulla transfer in 10 recipients

ES Injection

Mouse (chimeras not guaranteed).
Injection of min 30 C57Bl/6NCRL blastocysts per clone, other strains on demand, Uterustransfer in 3 recipients.
Alternativ: Injection of 60 Blastozysten per day, Uterustransfer in 6 recipients

Co-culture Chimeras

Co-culture of ES cells: Mouse (chimeras not guaranteed).
Aggregation of 60 C57Bl/6NCRL or FVB/N Morulae per clone. Other strains on demand.
Uterustransfer in 4-6 recipients

Cryopreservation of embryos

Cryopreservation of embryos in 2-cell stadium (Mouse and Rat)
Cryopreservation of 200 2-cells per strain/line in Minitübs of 20 2-cells aliquots.
Storage in two different buildings ander strictly monitored conditions in liquid nitrogen at -196°C (connected to central technical control room, on-call duty).)

Cryopreservation of sperm (Mouse)

Cryopreservation of sperm from 2-5 males of strain/line in 5 Minitübs per male.
Storage in two different buildings ander strictly monitored conditions in liquid nitrogen at -196°C (connected to central technical control room, on-call duty).)

Revitalization

Revitalization of embryos from the IBF embryobank of the BTL: Mouse and Rat

Thawing of 1-2 Minitübs approx. 20- 40 embryos (depending on strain the results of the embryos are at 90-98%. Generally 30-60% of transfered embryos are born. Oviducttransfer in 3-4 recipients.

Revitalization of embryos, which were not stored in the IBF embryobank: Mouse/Rat.

Embryos are thawed according to protocol, and transfered into recipients via embryotransfer. We can provide no guarantee, mostly due to unknown conditions of cryopreservation and storage. Very often also the case for quality and quantity.

Strain/line-sanitization (Mouse and Rat)

Collection of 2-cells from contaminated strain/line.
Oviduct (Mouse)/ampulla transfer (Rat) into SPF foster mothers (SPF Unit)
Mikrobiologal examination of foster mothers after weaning of pups

In Vitro fertilisation (IVF)

In-vitro-fertilisation mouse for “PROPAGATION“ of non breeding males: Mouse.

Collection of sperm from cauda epididymis
Fertilisation of oocytes from 15-20 C57BL/6NCRL females, other strains on demand.
Cultivation until 2-cell stadium
Oviducttransfer/Cryopreservation of developed 2-cells.

In-vitro-Fertilisation Mouse for the “PRESERVATION“ of several identical genotypes: Mouse.

Collection of sperm from cauda epididymis
Fertilisation of oocytes from 15-20 C57BL/6NCRL females, other strains on demand.
Cultivation until 2-cell stadium
Oviducttransfer of developed 2-cells.

If necessary, a laser-assisted IVF is going to be done.

Intracytoplasmatic sperm injection (ICSI)

ICSI: Mouse and Rat

Collection of sperm from cauda epididymis of living males (in case of dead males not after 12 hours post mortem)
Injection of spermheads into oocytes
Cultivation until 2-cell stadium
Oviducttransfer into recipients

Preimplantation diagnostics (PID)

PID: Mouse

Biopsy of zygotes in 4 t 8-cell stadium
Individual cultivation (until blastocyst) until diagnosis.
Uterustransfer of selected blastocysts

Lentiviral Transgenesis

LTG: Mouse/Rat

Injection, incubation of min 200 oocytes. Strains on demand.
Oviducttransfer into 10 recipients

Ovartransplantation (Mouse/Rat)

Collection of donor ovaries
Transplantation of donor ovaries in immunodeficient recipients
Mating until pregnancy

Computer Assisted Sperm Analysis (CASA)

after 14 days of sexual abstinence. Removal of both Cauda epididymis (CE), terminal or surgical
CE are immersed in 500µl pre-incubated M-HTF beneath paraffin oil
CE are opened (sperm float)
Computer assisted analyisi starts after 60 mins.