TIRF Station 2

Total Internal Reflection Fluorescence

Specifications

· Olympus IX 81 motorized inverted  microscope

· Olympus TIRF illuminator with fiber optic input

· Bioptechs 37°C environmental control also available.

· Condenser, 0.55 NA

· Objectives

10X UplanFl, 0.3 NA, PC, dry

40X UplanFl, 1.3 NA, oil, DIC-capable

60X Planapo, 1.45 NA, oil (TIRF lens)

· Argon laser line at 488 nm for TIRF

· HBO 100W illuminator for widefield  fluorescence

· Fluorescence filter sets  (6)

DAPI, FITC, TRITC, GFP

· MetaMorph  software, UCI

· Photometrics CoolSnap HQ CCD camera

· Equipment for microaspiration

Applications

· Widefield or TIRF fluorescence images of membrane proteins, receptor proteins, signalling molecules, cortical cytoskeleton proteins; TIRF excites fluorochromes within 100 nm of the coverslip surface

· Micromanipulators for holding micropipettes for aspirating cell surfaces to measure changes cell surface stiffness.

This microscope is made available to members of the microscope facility by Dr. Doug Robinson in the Department of Cell Biology

Example:  Myosin thick filaments in the cortex oflive Dictyostelium cells.