Leica TIRF

Brief description:

This epi-fluorescence microscope is additionally equipped with a TIRF unit enabling to view processes at the interface to the coverslip (adhesion, membrane fusion processes etc.). The system has a high speed function allowing for fast aquisition using the laser illumination. It  can also be used for automated longterm, multi-position life cell imaging. The setup includes a motorized X-Y stage, and an incubator box  temperature and CO₂). It is operated by Leica LAS AF software. An offline  version of the program exists on Image Processing Computer 1 in G 9.1 for viewing images and  further analysis (can also be downloaded onto your PC). The system is equipped also with an Eppendorf FemtoJet and InjectMan Ni2 microinjection unit, which allows semi-automatic microinjection of adherent cells.

Microscope:

• Leica DMI6000B (inverted), equipped for DIC and Phase contrast imaging

Ilumination:

• Leica Metal-halide illumination source (similar spectra like an HBO) for conventional widefield epifluorescence
• laser lines: 405, 488, 561 and 647 nm for TIRF

Objectives:

• 5x 0.12NA N-Plan
• 10x 0.25NA Ph1 N-Plan
• 20x 0.4NA Ph1 HCX PlanFluotuar LD (long distance)
• 20x 0.7NA HC PlanApo
• 40x 0.7NA Ph2 PlanFluotar
• 63x 1.4NA Oil HCX PlanApo DIC
• TIRF objective: 100x 1,47NA Oil HCX PlanApo 

Camera / detectors:

• Andor iXon EM CCD camera (front illuminated, 8 μm x 8 μm pixel size)

Software:

• Leica LAS AF software

Fluorescence settings:

• Widefield epifluorescence filtersets: DAPI , EGFP, Cy3, DsRed and Cy5