title: Self-interference fluorescence microscopy: three dimensional fluorescence imaging without depth scanning.
abstract: We present a new method for high-resolution, three-dimensional fluorescence imaging. In contrast to beam-scanning confocal microscopy, where the laser focus must be scanned both laterally and axially to collect a volume, we obtain depth information without the necessity of depth scanning. In this method, the emitted fluorescence is collected in the backward direction and is sent through a phase plate that encodes the depth information into the phase of a spectrally resolved interference pattern. We demonstrate that decoding this phase information allows for depth localization accuracy better than 4 µm over a 500 µm depth-of-field. In a high numerical aperture configuration with a much smaller depth of field, a localization accuracy of tens of nanometers can be achieved. This approach is ideally suited for miniature endoscopes, where space limitations at the endoscope tip render depth scanning difficult. We illustrate the potential for 3D visualization of complex biological samples by constructing a three-dimensional volume of the microvasculature of ex vivo murine heart tissue from a single 2D scan.
authors: Institute for Lasers, Life and Biophotonics Amsterdam, Department of Physics and Astronomy, VU University Amsterdam, de Boelelaan 1081, 1081 HV Amsterdam, The Netherlands. m.de.groot@vu.nl
authors: Mattijs
authors: M
authors: de Groot
authors: Conor L
authors: CL
authors: Evans
authors: Johannes F
authors: JF
authors: de Boer
mesh_terms: D000818:Animals; D006321:Heart / Q000033:anatomy & histology; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods*; D008857:Microscopy, Interference / Q000379:methods*; D055806:Microvessels / Q000033:anatomy & histology; D059629:Signal-To-Noise Ratio