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 less dense medium. In fact, the evanescent wave is used to illuminate the sample. Consequently, the possibility of observing a superficial (instead of bulk) part of fluorophore labeled sample is opened up. In this work, a total internal reflection fluorescence microscope based on the light guide has been designed and assembled by means of the inverted microscope to image a thin layer from the surface of the sample. Operated experimental arrangement has been employed for the total internal reflection fluorescence imaging of cadmium selenide (CdSe) quantum dots.