| | Nikon MicroscopyU: Fluorescence Microscopy - Total Internal Reflection Fluorescence |
 | | With adjustment of the laser excitation incidence angle to a value greater than the critical angle, the illuminating beam is entirely reflected back into the microscope slide upon encountering the interface, and an evanescent field is generated in the specimen medium immediately adjacent to the interface. |
 | | Total internal reflection does not occur suddenly as a new phenomenon at the critical angle, but a continuous transition is followed from predominant refraction with a small amount of reflection, to total reflection when the critical angle is exceeded. |
 | | Once the critical angle is surpassed, further increases in the radial distance of the laser focal point from the lens axis serve to reduce the evanescent field penetration depth in a smooth and reproducible manner. |
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