Wide-Field Two-Photon Fluorescence Microscopy

Abstract

The availability of lasers with femtosecond, ultrafast light pulses provides new opportunities and challenges in instrument design. This presentation addresses aspect of utilizing ultrafast light pulse in optical sectioning. The fundamental characteristics of fluorescence are introduced, and various fluorescence microscopies are explained in the view of the implementation by stating pros and cons. For wide-field 3D microscopy, the basic concept of temporal focusing is introduced, and a comprehensive mathematical model is derived for depth-resolved wide-field illumination by optimizing key instrument design parameters which enhances optical sectioning capability. Lastly, we achieve single nanoparticle detection and super-video rate imaging by employing high two-photon cross-section fluorescence probe.