Depth resolved wide field illumination for biomedical imaging and fabrication

Abstract

Nonlinear microscopic imaging is relatively slow due to the sequential nature of raster scanning. Recently, this limitation was overcome by developing a 3D-resolved wide-field two-photon microscope based on the concept of temporal focusing. The existing temporal focusing systems have poor optical sectioning capability and, due to a shortage of illumination power, low actual frame rate. In this presentation, a comprehensive mathematical model for temporal focusing two-photon microscope will be presented. By optimizing instrument design and the use of high two-photon cross section quantum dots, we demonstrate single quantum dot imaging with submicron resolution at video rate and applied it to study transport processes in cells. Further, we realize that the depth resolved wide field illumination can be used for microfabrication. A prototype three-dimensional lithographic microfabrication system was developed and micropatterning capability based on photobleaching process is demonstrated.