Optically driven full-angle sample rotation for tomographic imaging in digital holographic microscopy

Abstract

This study presents a novel tomographic imaging technique for living biomedical samples using an optically driven full-angle rotation scheme based on digital holographic microscopy, in which the three-dimensional refractive index distribution inside the sample can be measured and analyzed. To accomplish the full-angle sample rotation, two optical traps are driven by highly focused spots on the top and bottom of the sample. The rim image of the sample outside the focal depth at the different rotation angles and propagation distances can be corrected and compensated, respectively, via numerical focusing; therefore, tomographic imaging of the sample can be conducted. The proposed approach shows that an entire symmetric spectrum can be acquired for tomographic reconstruction without the missing apple core problem as in traditional sample-rotation schemes. The three-dimensional refractive index of living yeast in a fluid medium is measured and verified.

Tomographic diffractive microscopy: Towards high-resolution 3-D real-time data acquisition, image reconstruction and display of unlabeled samples

“…-TDM and its variants give access to the index of refraction distribution within the observed specimen, at the microscopic scale, and the interest of this non-labeling imaging modality has been widely demonstrated for biological investigations as well as for studying artificial structures [15][16][17][18][19][20][21][22][23][24][25][26][27][28]32,37,38,[41][42][43][44][45]. Note that the index of refraction is a complex quantity, related to the refraction and to the absorption.…”

“…2, right). Other approaches to fill the Fourier space exist, by rotating the specimen [15][16][17][18][19][20][21][22], changing the illumination wavelength [48,49], recording the reflected wave [39,40], or combining transmission and reflection [25].…”

“…While the precision of measurement of interferometric approaches is unsurpassed [12,13], the resolution when reconstructing 3-D transparent specimens is plagued by the small quantity of information, which is recorded, because of the use of only a single illumination direction [14]. In order to increase the recorded quantity of information, a rotation of the specimen [15][16][17][18][19][20][21][22], or a variation of the illumination wave inclination [23][24][25][26][27][28] can be used, and the set of recorded interferograms represents a diffractive tomographic acquisition [29,30]. This allows for a numerical 3-D reconstruction of the observed specimen.…”

High-Bandwidth 3-D Multitrap Actuation Technique for 6-DoF Real-Time Control of Optical Robots

“…Optical manipulation has become a popular tool for manipulating single biological samples, successfully demonstrated in a large range of in-vivo [3] and in-vitro [4] experiments such as the trapping of red blood cells in living animals [5], the immobilization of bacterial cells for nanoscopy [6] and cell rotation for tomographic imaging [7], among others. Usually, these optical traps are used directly to manipulate the object of interest.…”

Reconstructions of refractive index tomograms via a discrete algebraic reconstruction technique

“…These methods, however, require special setups and have difficulty in handling soft biological samples. Illumination rotation methods have also been presented using spatial light modulators, including galvanometric mirrors [15,17], liquid crystal spatial light modulators [18][19][20], and digital micromirror devices [21,22]. Still, however, the techniques suffer from the missing cone and the resultant anisotropic resolution.…”