Upgrade of Andor Dragonfly for Four Dimensional Cellular Medicine and Super-resolution microscopy

Here we are applying for an upgrade for the Andor Dragonfly spinning disk confocal/ super-resolution laser scanning microscope. We Beta-tested the Andor Dragonfly in 2016/17 and our system was the first in Europe. In the past 6 years it has made a significant contribution to the publication of over 22 papers. It has been used so much (>1000 hours per annum) in the ensuing six years that the stand and sCMOS camera, built around a Nikon microscope stand requires replacement. The system has carried out over 6000 hours of research in the past 6 years and is the busiest machine in the advanced imaging core facility. Since Dragonfly's original installation both Andor and Nikon have released new products that make the new Dragonfly system an even better fit to our research. We have been gifted an unused Nikon Ti2 stand which needs a Hardware autofocus, a laser and new objectives to be fitted. An automatic water dispenser, new silicone immersion objective and new highly sensitive camera (SoNa) will provide unparalleled depth of field and field of view to facilitate high resolution live imaging of organoids, model organisms and organotypics. To ensure that the system is well supported it will be housed in a dedicated live imaging suite and supported by expert technicians. We will keep the system under an all inclusive service contract. We are confident with the upgrades the Dragonfly will continue to be the cornerstone of our MRC HGU research. The Dragonfly is open access as part of the Edinburgh Super-resolution Consortium (ESRIC).

Dragonfly forms part of the Edinburgh Super-Resolution Consortium (ESRIC)/ BioImagingUK open access national imaging facility. Dragonfly is the cornerstone of the MRC HGU's 4D Cellular Medicine research strategy. It allows for fast, high-resolution analysis of 3D cellular models (tissue explants, organoids, embryos etc). Low photoxicity allows for high resolution live imaging, and the flexibility of this platform across scales embeds the ability to do super-resolution imaging in complex 3D cellular disease models. Here we are applying for funding to upgrade our existing system including replacement of the systems current stand with an upgrade. We already have a Nikon Ti2 stand it will need to be retrofitted with a perfect focus system, new objective lenses corrected in the violet and an automatic water dispenser, which will enable long term high resolution imaging of organoids, organotypics and model organisms long term. Since our system was purchased in 2017 Andor have brought our a new range of sCMOS cameras and a new laser system. Both of which will add to the current range of applications the system can address.

Our proposed upgrades would allow us to enable the following research from with in the MRC-HGU and IGC:
Super-resolution live imaging of dynamic cellular processes - gene regulation, autophagy, cilia, trafficking: (Bickmore, Mill, Wilkinson, Gilbert, Caraces)
Cell fate specification and lineage tracing: Patton, Long
Analysis of cell signalling in organoid disease and tumour models: Boulter and Jackson
Organotypic: Genetics of Eye function and dysfunction, and mechanisms of metastasis in bone: Bhartia, Megaw

Dragonfly has been used by other groups outwith the IGC as it is open access. It has contributed to 22 research papers at least and has been used for over 6000 hours of imaging since 2017.