Total Internal Reflection Fluorescence Microscope

This microscope is housed in BCMM 160. TIRF is achieved through 60X, 1.45NA or 1.49NA objectives and can be coupled with wide-field epifluoresence illumination via a 80/20 mirror located int he back of the microscope. Laser illuniation is provided by the Andor Revolution syste, which includes a 50mW/488nm, a 50mW/560nm and a 25mW/650nm solid-state lasers, shuttered via Acoustic-Optical Tunable Filters (AOTF). Images are acquired with a 512 x 512 back-illuminated, Electron-Multiplied (EM) CCD iXon 897 camera (Andor Technologies). Hardware and image acquisition are controlled via the Andor iQ imaging software, runnin on a Dell workstation.

Usage

TIRF microscopy provides selective illumination of the lower surface of the cells, near the glass/medium interface. Thus, it is possible to visualize processes that occur at the basa plasma membrane, such as endocytosis, exocytosis and cell adhesion, with unmatched signal-to-noise. Moreover, the availability of multiple laser lines enables simulataneous visualization of multiple markers of these processes. This is key to understanding how the variousl molecular components implicated in endocytosis are orchestrated, as well as how coordination between endocytosis and other biological processes at the surface of the cell is achieved.

Contact

For questions or to set up a time to use the TIRF Microscope, please contact Stacy Wilson at 737-4311 or stacy.wilson@yale.edu