In Vivo Imaging Facility
Two-photon fluorescence microscopy is a powerful research tool that employs ultra-fast pulsed lasers in combination with advanced optical laser scanning techniques to capture high-resolution three-dimensional images of fluorescently tagged specimens within intact tissues samples over time. This research methodology has the advantage of direct visualization of cellular arrangements and events that are often hard to deduce through sole use of in vitro assays and frozen sections, and the added benefit of being able to quantify many aspects of these events. As such, it is particularly well suited to address questions pertaining to the study of dynamic processes in living cells and tissues including, but not limited to, cell interactions, cell differentiation, and cell migration. Such studies can be conducted within virtually any tissue type; given tissue preparations can be developed to overcome the obstacles associated with maintaining native tissue structure and perfusion.
The In Vivo Microscopy Core Research Facility provides support for Yale investigators interested in utilizing two-photon fluorescence microscopy to achieve their research goals. The Core is responsible for novel protocol and methods development and is designed to serve as a clearinghouse for expertise and experience, being able to translate newly successful approaches to multiple projects and investigators. It is the Core’s goal to assist investigators in experimental design and implementation and to provide training for those whom wish to learn surgical preparations, imaging acquisition, and data analysis at the facilities image workstation utilizing Improvision’s Volocity software.
As an additional part of the educational outreach of this Core, we have, in collaboration with the Department of Cell Biology, put on annual hands-on Microscopy Workshops.
The imaging facility, which includes a dark imaging room and an antechamber for specimen preparations, is located in The Anlyan Center, in room S-614. The imaging workstation is located just down the hall outside of S-600.
In Vivo Administration
- Grants & Pubs
The In Vivo Imaging Facility’s microscope suite is comprised of an antechamber equipped with a sink, lab bench, and dissecting microscope for specimen preparation, and a dark imaging room that houses the microscope. Wall mounted inhalation anesthesia (isoflurane) is available in the dark imaging room to enable longer, safer imaging.
LaVision Biotec TriMScope
The facility upright, laser scanning, two-photon microscope is operated with a tunable (680nm – 1080nm) Titanium-Sapphire Laser (Chameleon Vision II, Coherent) purchased in January 2010. The microscope is outfitted with a water immersion, long-working distance Olympus 20X objective lens (NA 0.95) and is capable of imaging both fixed and live specimens. Currently, fluorescent emission from four colors (Far-red, Red, Green, Blue) can be detected simultaneously from a sample utilizing non-descanned photomultiplier tubes (PMTs).
Macintosh Computer Workstation
The Mac Pro facility computer workstation is located just down the hall from the microscope suite and is available for data analysis utilizing Imrovision’s Volocity software. Volocity utilizes a variety of tools that allow for measurements in both three and four dimensions and the generation of high-resolution images/movies.