Measuring incidence angle for through-the-objective total internal reflection fluorescence microscopy

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Measuring incidence angle for through-the-objective total internal reflection fluorescence microscopy.

Total internal reflection fluorescence (TIRF) microscopy has the exciting laser beam incident beyond critical angle from the glass side of a glass/aqueous interface formed by the coverslip and aqueous sample. The aqueous side evanescent field decays exponentially with distance from the interface with penetration depth depending on incidence angle. Through-the-objective TIRF has the exciting las...

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Fast high-resolution 3D total internal reflection fluorescence microscopy by incidence angle scanning and azimuthal averaging.

Total internal reflection fluorescence microscopy (TIRFM) is the method of choice to visualize a variety of cellular processes in particular events localized near the plasma membrane of live adherent cells. This imaging technique not relying on particular fluorescent probes provides a high sectioning capability. It is, however, restricted to a single plane. We present here a method based on a v...

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A Thin Layer Imaging with the Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy (TIRFM) is an optical technique that allows imaging of a thin layer of the sample with a thickness of about 100-200 nm. It is used in science of cell biology to study cellular processes, especially near the membranes of living cells. This method is based on the total internal reflection phenomenon, where the evanescent wave is generated in the l...

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Total internal reflection fluorescence (TIRF) microscopy.

Total internal reflection fluorescence (TIRF) microscopy (TIRFM) is an elegant optical technique that provides for the excitation of fluorophores in an extremely thin axial region ("optical section"). The method is based on the principle that when excitation light is totally internally reflected in a transparent solid (e.g., coverglass) at its interface with liquid, an electromagnetic field, ca...

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Topic Introduction Total Internal Reflection Fluorescence Microscopy

The goal in fluorescence microscopy is to detect the signal of fluorescently labeled molecules with great sensitivity and minimal background noise. In epifluorescence microscopy, it is difficult to observe weak signals along the optical axis, owing to the overpowering signal from the out-of-focus particles. Confocal microscopy uses a small pinhole to produce thin optical sections ( 500 nm), but...

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ژورنال

عنوان ژورنال: Journal of Biomedical Optics

سال: 2012

ISSN: 1083-3668

DOI: 10.1117/1.jbo.17.12.126007