Application Note

Application Note: Total Internal Reflection Fluorescence Microscopy

Source: Olympus America Inc.
Total internal reflection microscopy (TIRFM) is an optical technique used to observe single molecule fluorescence. Some biophysicists have used the technique for many years, while others are just beginning to explore the boundaries of this versatile mechanism for studying phenomena occurring at interfaces.

Today, the technique is gaining popularity with cell biologists and neuroscientists to employ it to observe cell membrane fluorescence, in part because new membrane-specific dyes have been developed. In the past, TIRFM was difficult to perform due to the complexity of the microscope setup and the problem of achieving acceptable image brightness. A recently developed high numerical aperture microscope objective lens that improves TIRFM and makes it widely accessible is discussed in this application note.