Tirfm extends confocal imaging capabilities

Combining confocal and Tirf imaging provides more comprehensive information about cellular processes than previously possible, by producing very clear images throughout the entire sample

Olympus Life and Material Science Europa has released a peerless total internal reflection fluorescence microscopy (Tirfm) module for its FluoView FV1000 confocal microscope.

The Tirfm module integrates easily with the confocal system.

For Tirf image recording the FV1000 microscope platform comes with the Olympus CellM/R imaging system including a high sensitivity CCD camera.

The Olympus FV1000 is described as the most flexible confocal system available, with features including the unique Sim scanner and superb UIS2 optics.

The addition of an advanced Tirfm module allows researchers to significantly enhance the imaging capacity of their system.

As well as standard and confocal laser scanning fluorescence image acquisition, the Tirfm module enables evanescent illumination for CCD-based thin surface layer observation.

It also provides higher Z-resolution than established light microscopy techniques.

The Tirfm module enables total internal reflection of the laser illumination within the glass of the slide or vessel below the sample.

This creates an evanescent wave which penetrates a thin surface layer of the sample, enabling very high signal-to-noise (S/N) ratio imaging of fluorescently labelled molecules in this layer.

The penetration depth, and thus the excitation of the fluorescent molecules, is achieved using the FV1000 scanner optics and can be user-set to a minimum of 50nm.

Complex time-lapse protocols such as sequential, multi-wavelength Tirf imaging, and variable penetration depth recording, can be easily programmed by the user.

The Tirf method is particularly useful for investigation of a wide-spectrum of cell surface events such as endo/exocytosis, cell adhesion and the formation of junctions between cells.

Combining both confocal and Tirf imaging in one complete system provides more comprehensive information about cellular processes than previously possible, by producing very clear images throughout the entire sample.

Using the Olympus FV1000 with Tirfm, the complete transport pathway of a protein can now be followed through to its exit from the cell in intricate detail with greatly reduced background noise.

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