Three-dimensional microscopy by optical scanning holography

Abstract

Luminescent markers play a key role in imaging techniques for life science since they provide a contrast mechanism between signal and background. We describe a new type of marker using second harmonic generation (SHG) from noncentrosymmetric BaTiO 3 nanocrystals. These nanoparticles are attractive due to their stable, non-saturating and coherent signal with a femtosecond-scale response time and broad flexibility in the choice of excitation wavelength. We obtained monodispersed BaTiO 3 nanoparticles in colloidal suspensions by coating the particle surface with amine groups. We characterized the SHG efficiency of 90-nm BaTiO 3 particles experimentally and theoretically. Moreover, we use the coherent SHG signal from BaTiO 3 nanoparticles for three-dimensional (3D) imaging without scanning. We built a harmonic holographic (H 2) microscope which records digital holograms at the second harmonic frequency. For the first time, high-resolution 3D distributions of these SHG markers in mammalian cells are successfully captured and interpreted by the H 2 microscope.

Optical sectioning for optical scanning holography using phase-space filtering with Wigner distribution functions

“…OSH is a recording technique that can record 3-D holographic information by using a single 2-D optical heterodyne scan [7]. It has been applied to many applications, such as 3-D microscopy [8] and 3-D pattern recognition [9]. In this work, we investigate a novel optical sectioning technique for OSH.…”

Autofocusing in optical scanning holography

“…The complex hologram of the object is recorded by adding the two outputs of OSH as in Eq. (2). Figures 4(a) and 4(b) show the amplitude and phase of the complex hologram.…”

“…Three-dimensional (3-D) imaging using optical scanning holography (OSH) has a long-standing history [1][2][3][4]. Most recently, OSH has been applied to 3-D microscopy [3][4][5][6], and an achieved resolution of better than 1 μm has been reported [5].…”

Principles and techniques of digital holographic microscopy

“…Instead of setting up interference of object and reference fields at the hologram plane, optical scanning holography (OSH) [211][212][213] illuminates the object with an interference field created by the overlap of two coherent beams, which is then scanned across the object (Fig. 29).…”