Project Summary
We request funds to support the upgrade of our existing Nikon A1R confocal microscope, which is now
almost 10 years old and lacks many of the newer features that have been developed since its purchase in
2012. The upgraded instrument will be housed within the Program in Membrane Biology (PMB) Microscopy
Core at the Massachusetts General Hospital (MGH) in the multidisciplinary Simches Research Center
building. The mandate of this MGH-approved Core facility is to provide access to a full range of microscopy
techniques to 2 NIH funded P30 Centers and individual research groups within the MGH community and
beyond. The Core has considerable expertise in static and live cell imaging techniques used by our
extensive user-base over the years. Our imaging capabilites were enhanced 4 years ago by the acquisition
of a Zeiss LSM800 Airyscan confocal platform to function alongside the older Nikon A1R. Together these
instruments have provided sufficient high level imaging bandwidth to serve our users. The Nikon A1R is
extensively used for longer term, live cell imaging and has an inverted microscope and an environmental
chamber to faciliate this need. The LSM800, associated with an upright microscope and no environmental
chamber, provides consistent and timely access to a high resolution confocal that is mostly used to perform
static imaging of fixed specimens. With the increase in demand for imaging of living cells expressing various
types of fluorescent molecules and tagged proteins, and the increasing need to image these cells over time
using 3D reconstructions, the purpose of this application is to provide our users with access to modern
confocal microscopy options that are better adapted for specialized imaging purposes than the current older
A1R system. In particular, the new high speed HD resonant scanning mode with associated AI image
processing capabilities provides a remarkable increase in image size and resolution when compafed to the
old resonant scanning system. Coupled with replacement of the old PMT detectors with new GaAsp
detectors and the incorporation of stable solid state lasers, these upgrades will allow specimens to be
exposed to much lower laser intensities, reducing photobleaching and photodamage, while providing a
significant increase in resolution compared to the old system. In addition, images and multicolor 3D stacks
can be collected much more rapidly in resonant scanning mode, while not sacrificing quality, meaning that
experiments can be conducted quickly and efficiently, saving time and money for our users. The PMB Core
has supported hundreds of peer-reviewed publications and dozens of grant applications over the years. The
Core has an ongoing quest to constantly improve and upgrade Core facilities in support of its user base.
This has been achieved using funds from NIH SIG applications as well as considerable institutional support
from MGH leadership. Providing our Core users with continued access to up-to-date confocal imaging is an
essential part of maintaining the MGH community at the cutting edge of NIH-funded, health-related research.