TIRF - MEDICAL IMAGING SYSTEMS III Total Internal...

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4/27/2011 Total Internal Reflection Fluoroscopy | Dhairya Pujara M EDICAL I MAGING S YSTEMS III Total Internal Reflection Fluoroscopy
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Medical Imaging System III TIRF Dhairya Pujara-12096058 Page 2 of 6 TIRF (Total Internal Reflection Fluroscopy) Introduction: Total internal reflection fluorescence microscopy (TIRFM) is an elegant optical technique mainly employed to observe single molecule fluorescence at surfaces and interfaces. The technique is often put to use for investigation of the interaction of molecules with surfaces, an area which is of fundamental importance to a wide spectrum of disciplines in cell and molecular biology. The very basic concept of TIRFM is simple, which mainly requires only an excitation light beam traveling at a high incident angle through the solid glass cover slip where the cells adhere. Light is refracted or reflected at the interface as a function of incident angle depending on the Refractive index differences between the glass and water phases. The beam of light is totally reflected from the glass/water interface at a specific critical angle. A very thin electromagnetic field (usually less than 200 nanometers) is generated by this reflection in the aqueous medium, which has an identical frequency to that of the incident light. The concept and physics of technology :
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Medical Imaging System III TIRF Dhairya Pujara-12096058 Page 3 of 6 Total internal reflection can occur when excitation light reaches an interface of a medium with a higher refractive index and a lower refractive index at an angle of incidence greater than a specific critical angle. When the light is totally internally reflected, it’s mainly because none of
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TIRF - MEDICAL IMAGING SYSTEMS III Total Internal...

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