Carl Zeiss TIRF

Carl Zeiss TIRF

Location: Medical Sciences Building, E-334

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  Unassisted Assisted
(1 hr)
UIC Cost $5/15/25* (per hour) $5/50/60 (per hour)
Other Academic $8/25/39 (per hour) $8/60/74 (per hour)
Non-Academic$10/30/50 (per hour) $10/65/85 (per hour)

Description

The Zeiss Laser TIRF imaging system is a microscope capable of visualising molecular level processes at or near cell membranes via the Total Internal Reflection Fluorescence (TIRF) principle. When properly configured, this imaging technique can result narrow imaging depths of less than 100 nm from a glass-specimen interface. (Note that TIRF requires a microscope objective with a high numerical aperture; the configured 100x/1.45 alpha Plan-Fluar is the only available option on the current system.)

The TIRF microscope is also outfitted with a complete Pecon XL TIRF S incubation system, which allows for the stabilization of temperature, CO2, and humidity with various stage inserts. These environmental variables can be controlled directly and easily through the AxioVision acquisition software to completement any live cell imaging approaches.

Even if the TIRF imaging is not feasible for a given sample, the system and its environmental controls can be used in transmitted light (bright field, phase contrast, differential interference contrast) or fluorescent imaging modes (the latter including reflectors for DAPI, CFP, GFP, YFP, and RFP-like signals). The microscope base features a fully motorized stage that, when initialized through the software, can scan preset areas of a single dish. The result is an ability to capture multiple time course experiments simultaneously from a single dish, which is well-suited for studies of cell migration, proliferation, or dynamic cell morphology.

Also available in the AxioVision software is the MosaiX module, an imaging processing algorithm that makes use of the motorized stage to stitch multiple images into a single seamless, tiled layout. Entire dishes or coverslips can thus be scanned and reconstructed as enormous, zoomable images.

*Given that the system can be used in separate transmitted light, widefield fluorescent, and laser TIRF imaging modes, different usage fees may apply.

More details can be found at the RRC Wiki guide for this instrument.

Technical Specifications

The available excitation lasers include:
• 100 mW tunable Ar laser = 458, 488, 514 nm (458 for CFP; 488 for GFP, CY2, FITC; 514 for YFP)

Currently configured objectives include:
10x/0.30 Ph1 EC Plan-Neofluar
20x/0.50 Ph2 Plan-Neofluar
40x/1.3 Oil DIC EC Plan-Neofluar
63x/1.4 Oil DIC Plan-Apochromat
100x/1.45 Oil DIC alpha Plan-Fluar

The system also includes a HAMAMATSU C4742-98-24ERG CCD camera, which is operated via the Axiovision 4.7 software package.