Photoacoustics with coherent light

Since its introduction in the mid-nineties, photoacoustic imaging of biological tissue has been one of the fastest growing biomedical imaging modality, and its basic principles are now considered as well established. In particular, light propagation in photoacoustic imaging is generally considered from the perspective of transport theory. However, recent breakthroughs in optics have shown that coherent light propagating through optically scattering medium could be manipulated towards novel imaging approaches. In this article, we first provide an introduction to the relevant concepts in the field, and then review the recent works showing that it is possible to exploit the coherence of light in conjunction with photoacoustics. We illustrate how the photoacoustic effect can be used as a powerful feedback mechanism for optical wavefront shaping in complex media, and conversely show how the coherence of light can be exploited to enhance photoacoustic imaging, for instance in terms of spatial resolution or for designing minimally invasive endoscopic devices. Finally, we discuss the current challenges and perspectives down the road towards practical applications in the field of photoacoustic imaging. (C) 2016 The Authors. Published by Elsevier GmbH.

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

Control of pulsed light propagation through multimode optical fibers

Edgar Emilio Morales Delgado

Visualization of organs and cells in the interior of living beings is a challenging task due to the light absorption and multiple light scattering occurring in biological tissue that prevents the di-rect transmission of images. A standard visualization approach is based on the use of endo-scopes which is accomplished through apertures in the body. Since their invention, the light transmission capability of optical fibers has enabled fiber optic based endoscopy with devices based on bundles of optical fibers that can directly relay images. Due to their small diameter, fiber optic endoscopes are standard in minimally invasive applications either with a white light illumination or as a fluorescent imaging device. The principle behind fluorescent endoscopy consists on the scanning of light intensity over a fluorescent sample, which is achieved by se-quential illumination of each one of the fiber bundles or by mechanical scanning of a single-mode double-cladding fiber. Collecting the fluorescent signal through the light guiding medium, an image can be reconstructed. The main drawback of fiber bundle endoscopes is the pixelated limited resolution given by the presence of thousands of cores that conform the bundle. In the double cladding fiber endoscopes, there is a limit to the miniaturization of the endoscope due to the requirement of mechanical elements at the tip of the fiber to scan the illumination over the sample. In this thesis, methods for focusing and digital scanning of optical light pulses through multimode optical fibers without any distal mechanical element are developed. Such techniques are based on spatial light modulation. Spatial phase modulation allows the control of light propagation in scrambling media, as in a multimode fiber, enabling the generation of intensity light foci or arbitrary intensity profiles. The high intensity of the transmitted pulses permits multi-photon imaging through multimode optical fibers, resulting in imaging probes of higher resolution than in the case of fiber bundles. We demonstrate in a working prototype that this approach provides all the advantages of multi-photon microscopy at the tip of an ultra-thin probe, such as a reduced photo bleaching of the sample, optical sectioning and enhanced image contrast. The developed methods can also be employed for material processing that requires high in-tensity light pulses. In specific, we demonstrate additive manufacturing, also known as 3D printing, at the tip of an ultra-thin needle. The working principle is based on two-photon polymerization, which is accomplished by scanning intense light pulses on a photosensitive material that hardens when exposed to light. All other additive manufacturing methods require very large nozzles or components in close proximity to the structure that they build, up to now. With an ultra-thin 3D printing probe we enable micro-fabrication through very small apertures, in places of difficult access or in the interior of living beings. We call it, endofabrication.

EPFL2017

High sensitivity, incoherent differential imaging

Chiara Bonati

Microscopic visualisation of optically transparent samples has been a topic of interest for several decades. Features such as density or chemical composition can influence the optical phase of transmitted light, and phase contrast can reveal these structures. Several methods of phase contrast have been developed, which can be categorised as either interferometric or non-interferometric, based on the type of coherence properties of the light used. In this work, I focus on incoherent based phase contrast, in particular Differential Phase Contrast (DPC). The choice of incoherent light brings benefits such as the absence of distortions like speckle patterns and ringing patterns, increased spatial resolution, and a simpler setup that can be used for in-vivo applications. Moreover, DPC allows reconstruction of quantitative phase maps of the samples. On the other hand, coherent-based techniques demonstrate superior performance in terms of phase sensitivity. The first part of this thesis offers a quantitative analysis of the phase sensitivity of DPC and investigates the influence of optical parameters and sample characteristics. With simulations and experiments, a relation between numerical aperture and phase sensitivity is demonstrated, and the concept of spectral matching is introduced to enhance the contrast. The methods can be generalised to any DPC setup, and allow a-priori investigation of the sensitivity of a DPC microscope at the design stage rather than through testing. Comparison between the best sensitivity that can be achieved in DPC and state-of-the-art interferometric techniques, shows that it is not possible to reach comparable single-shot performances. In this thesis, it is shown that the reason for this limitation is the strong background in DPC images, which degrades the dynamic range. DPC images are obtained with mirrored illuminations, for which the background is identical and the phase term switches in sign. The difference between these pairs of images is computed digitally, which does not improve the limited dynamic range. Lock-in DPC is proposed as a solution: instead of sampling different illumination states, lock-in DPC demodulates the phase signal when the illumination is switched, and the background is never encoded. This is enabled by periodical switching of sources coupled with a smart pixel detector, the so-called "lock-in camera". Part of this work is dedicated to the theoretical description of this method, and the analysis of the expected benefit. Experiments are also described, that demonstrate a factor of 8 improvement in single-shot sensitivity compared to standard DPC. DPC is not the only imaging technique to suffer from high intensity background: it is easy to see how the use of a lock-in camera for differential imaging can be generalised to any situation where weak modulations can be induced over a strong background. Here, an example is presented with lock-in Shifted Excitation Raman Difference Spectroscopy (SERDS). SERDS is an established Raman spectroscopy technique that takes advantage of the Raman emission spectrum being relative to the excitation wavelength to remove unwanted fluorescence emission. Two spectra with shifted excitation wavelengths are measured, their difference is computed, and the fluorescence is thus digitally removed. The parallel with DPC is immediately apparent. Both simulations and experiments are used to demonstrate the advantage of analog demodulation.

EPFL2022