Acquisition of Zeiss LSM980 with Airyscan 2, a super-resolution point scanning confocal microscope - PROJECT SUMMARY
Sanford Burnham Prebys (SBP) hosts labs with a very broad range of interests which include
cancer, aging and age-related diseases like neurodegeneration, cell and tissue homeostasis and
function, metabolism and stem cell biology. We truly believe that visualization of molecules of
interest, using dyes or fluorophores, in the context of both their spatial and temporal location within
cells or tissues is key to understanding physiological attributes of normal versus diseased states.
Super-resolution microscopy has become a key technology in providing this context. However,
an increase in spatial resolving power often requires a sacrifice in temporal resolution i.e. the
speed at which molecular kinetics of fluorophores can be observed, which necessitates careful
selection of the super-resolution mode required to fulfill experimental needs. For this purpose,
SBP requests support for a Zeiss LSM980 with Airyscan 2. The LSM980 is capable of
simultaneously imaging up to 4 different fluorophores with well-separated, non-overlapping
emission spectra with a two-fold increase in spatial resolution. The Multiplex modes, only
available on the LSM980, can speed up image acquisition up to twenty five-fold beyond
acquisition rates on typical point-scanning confocal microscopes, and still provide a near two-fold
increase in spatial resolution. This speed is essential in studying highly dynamic mechanisms that
require higher spatial resolution, such as vesicular trafficking, transient molecular interactions or
epigenetic changes. The ability to gauge subtle differences in these highly dynamic events will
give our users a better grasp of molecular changes that occur from healthy to diseased systems.
This speed of the Multiplex modes is also essential for fixed thick tissue imaging (>200µm) where
large fields of view or thick slices can be imaged in a fraction of the time, at super-resolution. The
LSM980 brings additional imaging benefits not currently available at SBP. The Quasar detector
permits simultaneous imaging of the full emission spectrum i.e. all fluorophores. The LSM980 is
more sensitive to weak samples and less sensitive to background noise, and the newly added
Airyscan joint deconvolution processing provides 90nm spatial resolution. These features make
it essential for projects requiring super-resolution imaging of thick samples, for e.g. organoids,
cell clusters or drosophila embryos. As the LSM980 will be the first available for the San Diego
community, the scientific advances made possible by acquisition of this instrument at SBP will
allow the 12 major users with 75% of the accessible usage time (AUT) to generate previously
unattainable data, which will further development of novel hypothesis, new funding and both
internal and external collaborations.