Cell Microinjection Centre
Microinjection is a powerful method that can be used to introduce almost any molecule (proteins, nucleic acids, cell extracts, small molecules, dyes, and drugs) into tissue culture cells. With some training, researchers can even inject specific subcellular compartments, such as the cell nucleus. This technique can be used to study how cells organize their three dimensional space, for example, how newly made mRNA is transported from the nucleus to its proper destination in the cytoplasm. Moreover, microinjection can be used to introduce foreign DNA into tissue culture cells in order to rapidly express genes of interest. DNA injection has many benefits over conventional transfection protocols in that 1) any tissue culture cell can be injected even difficult-to-transfect embryonic cells; and 2) microinjected cells express protein 1-2 hours post-injection whereas conventionally transfected cells take anywhere between 18-48 hours before the expressed protein can be detected.
Equipment
- Inverted Nikon Ti microscope stabilized on an air table and equipped with a Narishige hydraulic manual micromanipulator for somatic cell microinjection. The microscope also has a camera and monitor for instructional use.
- Epi-fluorescent inverted Nikon Ti microscope stabilized on an air table with both CCD 14bit and EMCCD cameras.
- Imaging computer equipped with Nikon NIS elements.
- Sutter needle puller for generating custom made microinjection needles.
- Table top preparative centrifuge for preparing microinjection samples.
- Tissue culture hood and incubator for maintaining microinjected mammalian tissue culture cells.
The video above describes an assay that employs the power of microinjection coupled with fluorescent in situ hybridization in order to accurately measure the nuclear export kinetics of mRNA in mammalian somatic cells.