Transgenic Core Equipment
Leica MZ6 microscopes have a 0.63x to 4.0x zoom range, 10x eyepieces are modular and can be adapted to accommodate a camera. The LCD light source is provided from above the working area and coarse or fine motor focus. The swing-arm configuration helps to expand the range of the stereo microscope and to move across a large work specimen. The core has two microscopes with the same configuration enabling simultaneous use for perform precision micro-surgery.
These stereo microscopes have a 0.63x to 4.0x zoom range, 10x eyepieces are modular and can be adapted to accommodate a camera. The light source is provided underneath the working stage. They are primarily used for micro-dissections and aid visualization of preimplantation stage embryos. They are a key component of the daily core work and have many uses.
Active vibration isolation tables with rigid steel frame constructed in a desk-style configuration. The core uses them for the demanding microinjection of pre-implantation stage embryos. They are designed to damper movements, that would otherwise not be noticed in typical laboratory settings. They are 3 ft x 4 ft and provide enough working space to accommodate an inverted microscope with manipulators.
These freezers provide a slow controlled rate of freezing in the range of -40oC to -80oC. The freezing time and temperatures can be adjusted for many applications. The core has two controlled rate freezers. The slow freezing protocol has proven to be very reliable and a consistent method for cryopreserving mouse strains.
These small benchtop incubators provide a microprocessor control system, humidified, gassed and stable heated environment for embryo culture. Due to their size and design they have the ability to rapidly recover homeostasis and temperature. The core has two MINC benchtop incubators and they are a crucial tool for performing temperature sensitive services such as in-vitro fertilization. They are strategically positioned next to the stereo microscopes and enable the user to warm and gas their micro-culture dishes faster than traditional CO2 incubators. They also require considerably less space, but do require a mixed gas combination, rather than just a CO2 supply.
This small easy maintenance storage unit has the capacity to store long insemination straws and cryovials that are attached to 12” canes. The canes sit inside one of six goblets and are stored in the liquid phase and have the potential remain viable indefinitely. The long neck of the canister reduces liquid nitrogen loss and improves vacuum performance. Samples in these units are maintained in liquid phase of nitrogen, rather than vapor-phase.
These large liquid nitrogen storage units have a large capacity, precise microprocessor control monitoring system, auto-fill system and a wide neck opening for easy access to all samples. The core has two units filled with custom made racks for housing cassettes and straws. Samples in these units are maintained in liquid phase of nitrogen, rather than vapor-phase.
The P-97 is used to pull glass capillaries for multiple applications, such as microinjection needles or holding pipettes. This instrument has many parameters, which can be adjusted, to create custom fine-tip needles for many specific research modalities. The core uses the P-97 puller to create its DNA/RNA microinjection needles and holding pipettes, used to secure a single embryo during the injection procedure.
The microforge offers a range of functions such as cutting, bending, fire-polishing and forming a spike. The thin filament can be heated and cooled as necessary. Rotation mechanisms allow the manual movement of glass capillaries. Positioning the capillaries near the heated filament will determine the shape and size of the end product. The core has a single micro-forge and routinely use it to greatly reduce the opening of pulled glass capillaries. These capillaries are then repositioned to produce a 20 degree bend approximately 5mm from the opening. The small opening prevents zygotes from being taken-up into the pipette and the smooth ends prevent cellular damage.
This high precision pneumatic manual microinjector is an air-controlled syringe-like piece of equipment. Once a glass capillary has been inserted into the instrument holder the user can position and secure zygotes, or blastocysts during the microinjection process. The core has two Cell Tram air microinjections, one for each microinjection station.
This high precision pneumatic manual microinjector is an oil-controlled syringe-like piece of equipment. Once a glass capillary has been inserted into the instrument holder the user can select healthy single embryonic stem cells. The system will hold the cells inside the capillary until ready to inject them into blastocysts. The core has two Cell Tram oil microinjections, one for each microinjection station.
The manually adjustable inverted microscope is designed for transmitting light to aid visualization of mouse sperm, oocytes, zygotes, blastocysts, and ES cells. The core has two inverted microscopes positioned on top of TMC anti-vibration tables. Each scope is equipped with 4x, 10x and 20x DCI optics enabling high magnification of samples. The microscopes are used to support and visualize embryos during the microinjection of CRISPR, DNA or ES cells.
The laser is attached to one of the inverted microscopes and is used for Assisted-IVF to create a hole in the zona pellucida of each oocyte, thereby facilitating sperm entry and fertilization.
These are considered the ‘arms’ of the microinjection process. They control the fine movements that are a critical component for manipulating and microinjecting zygotes or blastocysts.
The FemtoJet with its built-in compressor provide regulated pressure to expel DNA transgenes or CRISPR reagents through injection needles into zygotes during microinjection.
This electroporator uses capacitators to produce controlled exponential or square wave electrical pulses for cell penetration. This piece of equipment is used to electroporate CRISPR RNP products into zygotes to create new founder strains.