TIRF Station 1

Specifications

· Olympus IX 71 inverted  microscope

· Olympus TIRF illuminator with fiber optic input

· Bioptechs 37°C environmental control equipment available.

· Condenser, 0.3 NA

· Objectives

10X UplanFl, 0.3 NA dry

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

100X, 1.65 NA, oil (TIRF lens)

· 50 mW Kr/Ar laser lines available for TIRF with Prairie Technologies laser launch:

457, 488, 514, 568, 647 nm

· HBO 100W illuminator for widefield  fluorescence

· Fluorescence filter sets  (6)

CFP, GFP, YFP, dsRed, CFP-YFP, GFP-dsRed

· MetaMorph  software, UCI

· Photometrics Cascade 512B intensified CCD camera with on-chip multiplication gain, >90% QE, and 8- to 16-bit processing for single molecule imaging

Applications

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

· Single molecule detection at ~30 frames/sec

· On-chip multiplication gain of camera greatly boosts weak signals and shortens required exposure times

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

Example:  Vinculin in HEK cells, wide-field and TIRF