LEAP: Laser Enabled Analysis and Processing
Provides high-throughput microplate imaging of live cells in-situ, with laser-mediated cell processing capabilities
We are one of the few places in the UK with access to the versatile LEAP technology, which provides high-throughput microplate imaging of live cells in-situ, with laser-mediated cell processing capabilities. Cell populations can be selected by user-defined parameters, allowing elimination of unwanted cells, and optoinjection of a wide range of macromolecules into specific cell populations.
Brightfield imaging is LED-based, while multicolour fluorescence utilises a hybrid-halide lamp with 8 excitation and 8 emission filters, coupled to a CCD camera. Magnification is adjustable with 3x, 5x, 10x, and 20x available. Multi-wavelength imaging is integrated with software allowing cell characterisation and population selection by fluorescence intensity, size, granularity and shape.
Pulsed lasers are used only in cell processing (no imaging), with 2 wavelengths: 355nm (UV) & 532nm (Green). Laser targeting can be automated or manual, shooting up to 1000 cells/sec.
In-well elimination of unwanted cells with minimal effect on remaining cells has been used in fast and efficient selection of transduced gene expression, and in the generation of high-secreting cell clones. A range of mechanisms can be used in cell removal. Photothermal purification leads to immediate cell necrosis and protein coagulation (minimising cell contents leakage), Photomechanical purification leads to immediate cell lysis, and photochemical processing to targeted cell apoptosis over 4-24h.
Laser-mediated optoinjection is particularly useful for difficult-to-transfect cell types, including primary cells. Pulsed laser targeting leads to transient cell permeabilisation, allowing delivery of a variety of macromolecules including plasmid DNA, siRNA, peptides and small proteins. Off-target effects are reduced with minimal effects on cell viability, compared with lipofection and electroporation.
Generates identically-sized embryoid bodies without the use of collagenase or other enzyme dissociation. Photothermal “cutting” of cell layers minimises cell loss. This technique has been shown to maintain a high level of downstream differentiation potential.
Acknowledging the contribution of our research facilities provides a mechanism to demonstrate their success thus increasing their visibility internally and externally. In turn, this generates additional income to the School through external collaborations, it raises our profile within the UK and internationally, and it strengthens our funding applications and REF cases.
Please routinely note the contributions made by research facilities and staff in all publications.
As a matter of courtesy, and for your own benefit, please send a copy of the draft publication to the facility prior to submission. Facility staff can help spot errors in methods or presentation of results, and it provides an opportunity for them to ensure you have acknowledged them correctly.
Please communicate with, and acknowledge, specific facility staff if they were involved in performing experiments or provided more involved training/advice/work for you. Please make sure you consider co-authorship if their contributions go beyond this.
Standard text to include:
Cell processing, analysis and optoinjection were carried out using LEAP Technology supported by the UK Centre for Mammalian Synthetic Biology (funded by BB/M018040/1).
If you think the LEAP platform could assist with your research goals, please contact Dr Gaynor Campbell to discuss your project.