Selective excitation light fluorescence (SELF) imaging

Abstract

Fluorescence imaging is a potential candidate for tissue diagnostics in a wide variety of clinical situations. In order to extract diagnostic information using fluorescence, different approaches may be used. Typically, fluorescence imaging is performed by illuminating the sample at a single excitation wavelength and detecting the emissions at one or more wavelengths. We have built a prototype system for a new fluorescence imaging technique denoted Selective Excitation Light Fluorescence (SELF) Imaging. In this technique, the sample is illuminated with multiple excitation wavelengths, and one or more emitted wavelength images are detected. By using a multitude of illumination wavelengths or a weighted sum of illumination wavelengths, SELF imaging can highlight differences in the excitation spectra of fluorophores in the sample. Some potential advantages of this imaging technique are: detection of multiple labeled objects in microscopy using only a single filter cube, increasing the number of simultaneous labels which can be used on a single slide as labels are separated by their absorption spectra not just their emission spectra, detection of different components of tissue based on different excitation spectra, etc.