Project Summary
The Salk Institute for Biological Studies is an independent, non-profit research institute in La Jolla, CA that
focuses on fundamental basic science research. Shared resources and cores provide critical support for this
research. Eleven technology cores serve the Institute, including the Waitt Advanced Biophotonics Core Facility,
which is solely dedicated to technologies relating to multiscale light and electron microscopy imaging
and analysis. This Core has a significant dependency on confocal microscopy. Confocal microscopy is a
powerful research technology that is heavily utilized by NIMH-supported Salk scientists. In recent years,
usage of core confocal microscope systems has increased significantly, and the ability to accommodate all
requests has been possible only by operating all three existing confocal microscopes at near maximum
capacity. However, the Zeiss 710 is over a decade old. This instrument has limited technical capabilities,
decreasing reliability, and is near the end of its practical lifetime. The purpose of this grant application is to
address a pressing and unmet need for a replacement instrument, by the acquisition of an Olympus
FV3000RS TruSpectral Hybrid scanning confocal microscopy system. This new instrument would replace the
aging Zeiss 710, which will be placed on the resale market, donated to a non-profit organization, or recycled.
The FV300RS is a best-in-class instrument, ideally suited to the demanding imaging needs of Salk NIMH-
supported researchers and others. It offers a vast improvement in imaging technology that meets the more
advanced needs of the field. The system as configured has 7 excitation laser lines and unique holographic
spectral separation technology that offers real-time 16-channel spectral separation with 2 nm resolution and
very high light transmission. This improves signal to noise and offers true spectral un-mixing for challenging
multispectral imaging needs. In addition, it has bi-modal scanning capabilities, allowing either super
resolution precision laser scanning or very fast resonant laser scanning. It has 4 high sensitivity GaAsP
photomultiplier detectors and is equipped with the new Olympus X-line objectives that offer unparalleled
chromatic correction and image flatness without sacrificing high numerical aperture or large fields of view.
For challenging 3D multispectral imaging of large tissue volumes that are needed for whole brain connectome
and cell identity studies, this system has been configured with Olympus’s unique silicon immersion 40X
objective. This is specifically designed for whole brain cleared tissue imaging. As configured, the
FV3000RS TruSpectral Hybrid scanning confocal microscopy system offers flexible macro-to-micro scale,
rapid and super-resolution multidimensional imaging of fixed and live samples and provides a complete imaging
solution for the demanding needs of current and future neuroscience research. With the experience and
expertise provided by the Waitt Advanced Biophotonics Core facility, this new instrument will be immediately put
to a high level of use to facilitate scientific discovery.
