Two-photon imaging using a flexible endoscope

摘要

Two-photon autofluorescence imaging offers the analysis of cells and tissues without the need of taking biopsies, staining and complicated confocal detection systems. Therefore, it is of special interest for non- or minimal invasive clinical diagnostics. Until now, two-photon imaging was performed only on superficial surfaces like skin or of biopsies. To extend this technique to deeper tissues or inside the body the optical properties have to be reduced to endoscopical sizes. This can be achieved by tiny GRIN-optics, based on a radial gradient in the reflective index. A newly developed GRIN-lens assembly with increased numerical aperture is of special interest which is shown by the quality of tissue constituents and cell autofluorescence images. A fiber directs the laser light to the specimen in an assembly like an endoscope. This well-characterized photonic crystal fiber supports the high laser power of the femtosecond excitation impulses without the generation of non-linearities. A sensitive PMT detector detects the fluorescence. First fluorescence images using a fiber-GRIN lens combination were taken.