Micropipette/microaspiration (cell mechanics)
Total-internal Reflection Fluorescence Microscope (TIRF)

Model Information

• Olympus IX81 motorized microscope

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: 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/micropipettes for deforming cell surfaces by quantitative micro-aspiration, allowing measurements of cell cortical 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 of live Dictyostelium cells.