Optical Diffraction Tomography Using Nearly In-Line Holography with a Broadband LED Source

Ahmed Ayoub, Demetri Psaltis, Abhijit Roy
MDPI, 2022
Journal paper

Abstract

We present optical tomography methods for a 3D refractive index reconstruction of weakly scattering objects using LED light sources. We are able to record holograms by minimizing the optical path difference between the signal and reference beams while separating the scattered field from its twin image. We recorded multiple holograms by illuminating the LEDs sequentially and reconstructed the 3D refractive index reconstruction of the sample. The reconstructions show high signal-to-noise ratio in which the effect of speckle artifacts is highly minimized due to the partially incoherent illumination of the LEDs. Results from combining different illumination wavelengths are also described demonstrating higher acquisition speed.

Official source

https://infoscience.epfl.ch/record/292692?ln=en

About this result

This page is automatically generated and may contain information that is not correct, complete, up-to-date, or relevant to your search query. The same applies to every other page on this website. Please make sure to verify the information with EPFL's official sources.

Related concepts

Related publications

Optical Coherence Tomography (OCT) is a recent biomedical imaging technique based on low-coherence interferometry that is capable of acquiring depth-resolved reflectivity maps of scattering tissues with high sensitivity. The conventionally employed imaging mode is to build up a cross sectional image by scanning the sample surface with a point illumination. In order to increase imaging speed, the concept of parallel detection has been introduced to OCT. Here, a whole sample line or even surface is imaged directly onto an array of photodetectors, making the lateral scanning motion obsolete. A customized detector array has been developed in our institute for parallel OCT imaging based on complimentary metal-oxide-semiconductor (CMOS) technology. It associates a signal processing circuit to each photosensitive area of the array, capable of demodulating the interferometric signal on-chip. In this manner, only the signal envelop has to be read out, allowing for a high dynamic range and high frame rates. At the beginning of the thesis, this device had only been tested for topographic measurements on reflective surfaces. As a direct continuation of this previous work, the initial objective of this thesis has been to extend its application to parallel OCT in scattering samples and to identify other uses of this technology in the field of OCT. Accordingly, the first part of this work is dedicated to parallel detection in OCT, using this customized CMOS detector array. The feasibility of the approach is shown experimentally on scattering samples. Reflective surfaces covered with scattering solutions of varying concentration and onion samples are studied. Furthermore, the initial goal of fast OCT imaging is pursued by using the detector at its technological limits and realizing video-rate, three-dimensional OCT acquisitions of a dynamically changing sample. The thermal deformations induced by the probing beam on a dark strand of human hair are imaged at 25 volume acquisitions per second. Identified as a possible new application for the CMOS detector array, the concept of wavelength de-multiplexing for spectroscopic OCT is investigated in the second part of this thesis. Wavelength de-multiplexing is an experimental method for realizing spectroscopically resolved, time-domain OCT measurements. In contrast to the currently employed numerical post-processing methods for extracting spectroscopic information, wavelength de-multiplexing relies on optical wavelength separation in the interferometer detection arm and acquisition of independent wavelength channels using a detector array. The method itself is first studied using a laterally translated photodiode for detection, in order to compare it to the conventional spectroscopic OCT approach. The absorption characteristics of a glass filter and a Nd-doped crystal are measured in this manner. Then, associated to the CMOS detector array in a proof-of-principle experiment, the dynamically changing, spatially resolved absorption of a dye mixing process is measured online. The experimental setup used for wavelength de-multiplexing has lead to an innovative technique that is of interest to low coherence interferometry in general. The axial depth scan employed in all time-domain low coherence interferometry setups should ideally introduce a linear change with time of optical path difference between the interferometer's sample and reference arms. However, depending on the scan method used, the path difference variation is only approximatively linear and can even vary randomly from scan to scan. For precise, phase-sensitive and repetitive measurements the scans have to be calibrated. By increasing the coherence length of part of the detected radiation in the detection arm of the interferometer through narrow spectral filtering, a calibration signal is created that permits the precise measurement of the optical path difference variation and thus its calibration without the need for a secondary interferometer. The usefulness of the approach is demonstrated by combining it with a recently published spectral shaping technique for sidelobe suppression in OCT when using non-Gaussian source spectra. In order to be beneficial, such a technique requires highly repetitive measurements that can only be obtained with particularly stable scanning devices or well calibrated ones. In summary, this thesis studies three innovative experimental concepts that aim to improve OCT imaging speed and imaging quality and proposes them as interesting new tools to the OCT community.

EPFL2004

Multiple scattering in wide-field optical coherence tomography

Boris Karamata

Optical Coherence Tomography (OCT), a well-established imaging method based on low-coherence interferometry, provides cross-sectional images of the internal structure of biological samples with a resolution in the micrometer range. OCT was successfully applied on various tissues such as for instance the retina, the skin or a tooth. In highly scattering tissues like the skin, probing depth is limited to approximately 2mm, mainly due to insufficient rejection of multiply scattered light. Presently, the contribution of multiple scattering in OCT is not fully understood. Therefore, there is a strong and urgent need to develop models allowing a reliable evaluation of the system's limitations as well as the improvement of the imaging capabilities. It is generally believed that a relevant model should account for loss of correlation between the reference and the sample field due to multiple scattering. We developed a new comprehensive model of OCT. Our preliminary study revealed that the reference and sample fields are actually fully correlated. This important result allowed us to model the OCT signal as a sum of stationary random phasors and treated it as a statistical signal. The mean of this signal can be calculated thanks to classical results of statistical optics and to a Monte Carlo simulation. Unlike other existing models, our model accounts for the source autocorrelation function. The model proved to be in excellent agreement with a whole range of experimental data gathered in a comprehensive study of cross-talk in wide-field OCT. Moreover, our results put in question the applicability of widely used models of OCT based on the "extended Huygens-Fresnel principle", which assume a partial correlation between interfering fields due to multiple scattering. The construction of conventional OCT images is based on lateral scanning of a beam focused within the sample. To increase image acquisition speed and eliminate the need for lateral scanning, wide-field OCT was recently developed. Our experimental and theoretical investigations of the potential and limitations of wide-field OCT revealed the crucial role played by the spatial coherence of the light source. Spatially coherent illumination generates considerable coherent optical cross-talk, which prevents shot-noise-limited detection and diffraction-limited imaging in scattering samples. The dependence on several parameters of the optical system and of the sample properties was investigated in a comprehensive study. Cross-talk increases with the wide-field diameter, numerical aperture, source coherence length, and sample optical density; and strongly depends on sample anisotropy. We showed that spatially incoherent illumination realized with a thermal light source permits cross-talk suppression in wide-field OCT, i.e. rejection of multiply scattered light to a level comparable to that of point scanning OCT. We performed a theoretical study which revealed that the power per spatial mode radiated by thermal light sources is too low to permit a high signal-to-noise ratio while maintaining a fast acquisition speed. Therefore, wide-field OCT realized with either spatially coherent or spatially incoherent illumination suffers from inherent fundamental limitation. This led us to investigate the possibility of exploiting a spatially incoherent light source brighter than a thermal light source. We came to the conclusion that such a "pseudothermal" light source can potentially lead to wide-field OCT systems devoid of cross-talk and an image acquisition speed higher than that of a point scanning OCT system. However, the attractive properties of pseudothermal light sources could be gained at the expense of the simplicity and the economical advantages offered by thermal light sources. Furthermore, fast acquisition speed also relies on a performing "smart pixel detector array". Presently, such detectors do not have sufficient sensitivity and their frequency readout is too low as shown in our feasibility study. We performed a theoretical investigation of the potential of thermal light sources in terms of axial resolution and power per mode. The former revealed that the maximum power per mode is radiated at a wavelength higher than the spectral peak of a blackbody radiator. This led to the important practical conclusion that, at 6000 K, the maximum power is collected in the therapeutic spectral window in OCT (600 - 1300 nm), while at 3000 K this peak is shifted out of the therapeutic window leading to significant power losses. More generally, our work provides a design tool for choosing the optimal thermal light source for a given therapeutic window in terms of signal-to-noise ratio. Currently available sources at 6000 K consist of high pressure gas arc lamps providing a spectrum endowed with spectral lines deleterious for OCT. By suppressing a portion of the spectrum devoid of spectral lines of a mercury arc lamp, we obtained amongst the highest axial resolution so far reported in OCT. Furthermore, the importance of the speckle statistics in OCT incited us to clarify the origin of a difference between two theoretical results reported in the literature. Indeed, two calculations of the amplitude distribution of speckles in OCT, each of them based on a different mathematical formulation, yield different results. We showed that a modification of an initial assumption in one of the formulation leads to equivalent results. In conclusion, this thesis provides a deeper understanding of the potential and limitations of widefield OCT, leading to important design rules. Moreover, it presents a new comprehensive model of OCT putting in question other widely used models.

EPFL2004

Optical microscopy for imaging through scattering media

Xin Yang

Optical microscopy has been widely used in the life sciences and biological research for several decades. It possesses many advantages over x-ray radiography, computed tomography, ultrasound imaging and magnetic resonance imaging, in terms of being low cost, high resolution, harmless and capable of multiple contrast mechanism, etc. However, the weakness of optical microscopy is the inability to perform deep tissue imaging. Owing to the short wavelength of optical waves, it experiences much more scattering than longer waves in inhomogeneous materials such as biological tissues. Scattering scrambles the wavefront and limits the penetration depth of optical microscopy to only hundreds of microns. We explore in this thesis, novel optical approaches to image through scattering media. There are two main categories of the state of art techniques: imaging with ballistic photons, which requires separation of ballistic photons from the sneak or multiple scattered ones; and imaging with scattered photons, which relies on manipulation of the scattered photons according to the specific scattering properties. We primarily focus on the second category in this thesis. Taking advantage of the reversibility of light, phase conjugation has a long history in aberration compensation. A deterministic wave is incident on a scattering medium and results in a random scattered field. If we record the full information of the scattered field (amplitude and phase) and generate a phase conjugated replica of it, the phase conjugated wave can follow the same path and reconstruct the original deterministic wave after passing back through the scattering medium. In this work, phase conjugation is achieved using digital method, called digital phase conjugation. The scattered field is recorded through digital holography, and a spatial light modulator is applied to construct the phase conjugated wave. Second harmonic radiation imaging probes are used as the original light source, rendering the generation of a focus behind the scattering medium. The memory effect, which indicates the correlation between speckles with different incident angles, is exploited to move the focus around and obtain three-dimensional scanning microscopy behind scattering medium. Speckle scanning microscopy is the second method demonstrated in this thesis for imaging through turbid media. In contrast to digital phase conjugation, which requires access from both sides of the scatterers to measure the scattered field, in speckle scanning microscopy, excitation and collection can be achieved from the same side. The specific scattering events are not taken into account, instead, the statistic property of the scatterers plays a major role. This technique makes full use of two facts: 1. The statistically averaged autocorrelation of speckle fields approaches a delta function; 2. When a speckle pattern scans across a fluorescence object, the collected fluorescence intensity is proportional to the convolution of the speckle field and the object. Linking these two facts with delicate calculation, we can eventually minimize the influence of the speckle, and reconstruct two-dimensional images of objects behind turbid medium. Encouraged by the success of two-dimensional speckle scanning microscopy, we investigate more advanced possibilities by simulation, including three-dimensional speckle scanning and reference based speckle scanning techniques. [...]

EPFL2014

Multiple scattering in wide-field optical coherence tomography

Boris Karamata

EPFL2004

Optical microscopy for imaging through scattering media

Xin Yang

EPFL2014

EPFL2004