We propose a B-scan-wise- multi-focus average method (B-MFA) to suppress multiple scattering noise for in-vivo imaging. This method uses only multiple B-scans with modulated focus position for complex averaging, and a one-dimensional computational refocusing is applied to cancel the defocus in each B-scan. To validate the proposed method, ten medaka fish were measured in-vivo without anesthesia. The results show that B-MFA can enhance the OCT image contrast in the deep regions.
We demonstrated dynamic optical coherence tomography (D-OCT) to human skin in vivo by applying a sample fixation attachment and bulk motion correction algorithm to correct the motion artifacts. A D-OCT contrast of logarithmic intensity variance (LIV) was calculated. Without sample fixation attachment and the motion correction algorithm, the whole image area exhibited high LIV, and no meaningful structure was seen. The application of the motion correction methods revealed fine en face vessel structures, which cannot be seen in OCTA. The statistically significant motion artifact reduction capability of our motion correction method was also shown by t-test.
Dynamic optical coherence tomography (DOCT) is a label-free technique that visualizes tissue dynamics by analyzing a long-time sequence of OCT images. Although it was successful for in vitro and ex vivo imaging, it is still challenging for in vivo imaging because of the sample motion. We address this issue by developing hardware- and software-based motion suppression methods and demonstrating in vivo DOCT imaging of human skin. The hardware method is a sample fixation spacer. The software method is an image-registration based motion correction. We used logarithmic intensity variance (LIV) method to image the tissue dynamics. LIV was calculated from 32 sequential OCT frames taken within a 6.35s time window. The interframe time interval was 204.8ms and the entire DOCT volume was measured in 52.4s. Furthermore, we measured OCT angiography (OCTA) by standard raster scan with four frame repeats. To quantitively analyzing the improvement of proposed methods, three regions of interests (ROIs), each measuring 176μm (in depth) × 217μm (in lateral direction), was select from one B-scan to calculate the mean LIV. The improvement was assessed by paired t-test. The motion correction methods significantly reduced the high-LIV artifacts and revealed very fine capillary structures that had been buried by motion artefacts. The paired t-test results showed that the combination of fixation spacer and the software correction significantly reduced LIV artifacts (p=0.0052, 0.0137 and 0.0068 for three ROIs).
We demonstrate the label-free intra-tissue activity imaging of human induced pluripotent stem cell (hiPSC)-derived alveolar organoid including normal and fibrosis model with three-dimensional Dynamic Optical Coherence Tomography (D-OCT). The volumetric D-OCT data were obtained by a 840-nm spectral domain OCT. And the D-OCT images were obtained by “logarithmic-intensity-variance (LIV)” method, which is sensitive to the magnitude of tissue dynamics. Cystic structures with high-LIV border and mesh-like structures with low LIV were observed in all the organoids. They correspond to alveoli and fibroblast. The LIV images also visualized the abnormal repairing process of the alveolar epithelium.
KEYWORDS: Optical coherence tomography, In vitro testing, 3D modeling, Visualization, Tissues, Stereoscopy, Medical research, Lung, Drug development, Cancer
We demonstrate high-resolution (3.8-µm axial and 4.8-µm lateral) three-dimensional dynamic (D-) OCT imaging by 840-nm spectral domain OCT, and compare it with a low-resolution 1.3-µm D-OCT. The D-OCT images are obtained by logarithmic-intensity-variance method, which is sensitive to the magnitude of signal fluctuation.
Human-induced-pluripotent-stem-cell derived alveolar (lung) organoids and human breast cancer (MCF-7) spheroids were examined. The high-resolution D-OCT revealed the tessellation of high and low dynamics at the matured alveolar epithelium. It is also found that such matured alveolar epithelium exhibits ragged inner surface. For the spheroids, high-scattering spots with low dynamics were observed only in the high-resolution image.
KEYWORDS: Optical coherence tomography, Scattering, Multiple scattering, Glasses, Particles, Tissues, In vivo imaging, Image acquisition, Computation time, 3D acquisition
In this study, we propose a B-scan-wise-multi-focus averaging method (B-MFA) to suppress the multiple scattering noise in optical coherence tomography. This method uses multiple B-scans with different focus positions. These B-scans were one-dimensionally computationally refocused and complex averaged. To verify the multiple scattering noise suppression ability of this method, we measured a phantom consisting of scattering and non-scattering layers. The results show that BMFA suppresses multiple scattering noise.
Optical coherence tomography (OCT) has been utilized to investigate the optical properties of tissue in animal model studies. Polarization sensitive OCT (PS-OCT) is one of the functional extensions of OCT, which provides additional contrasts such as birefringence and degree-of-polarization uniformity (DOPU). It enables multi-contrast imaging for discrimination of tissue properties in biological samples. However, biological samples often exhibit multiple scattering (MS). It severely hampers the penetration depth and degrades the optical property measurement in deep regions for both OCT and PS-OCT. In a recently presented work, it was found that all of the scattering OCT value, birefringence value, and DOPU value were altered if they were measured at the deep region. This alteration is suspected to be artifacts caused by MS. So far, various methods have been proposed to tackle the MS issue. One is averaging several images with decorrelated MS signals. The decorrelation of MS signals was realized by wavefront shaping with complex devices such as spatial light modulators or deformable mirrors. However, these optics increase the cost and system complexity, and the wavefront shaping is still challenging. Simple hardware, such as multimode fiber, was only demonstrated in full-field OCT. In this work, we present a method termed as “multi-focus averaging” (MFA) to suppress the MS effect and improve the image contrast in deep regions of scattering samples. It uses focus modulation by a tunable lens to decorrelate MS signals among multiple acquisitions, and computational refocusing is used to cancel the different defocus during focus modulation. The refocused OCT signals are averaged in complex to reduce MS signals. It is demonstrated in standard OCT scheme, and a scattering phantom and a post-mortem zebrafish are used to validate the intensity contrast improvement. Results show that MFA images can provide better intensity contrast than standard complex averaging in scattering samples. In addition, we further adapt this method to PS-OCT to suppress the MS and to improve the accuracy of polarization sensitive measurement. We found MFA mitigated low-DOPU artifact in polarization sensitive measurement.
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