<i>In vivo</i> identification of arteries and veins using two‐photon excitation elastin autofluorescence

Li, Hui; Yan, Meng; Yu, Jia; Xu, Qianfan; Xia, Xianyuan; Liao, Jiuling; Zheng, Wei

doi:10.1111/joa.13080

Cited by 15 publications

(8 citation statements)

References 31 publications

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“…Blood vessels would also have good contrast with this modality and form a network around adipocytes. Elastin fibers are key components of blood vessel architecture and exhibit TPEF in the spectral range imaged here [ 23 ]. We verified that the TPEF signal was neither delay dependent nor did it reduce when the Stokes was blocked so it is not a four-wave mixing signal.…”

Section: Resultsmentioning

confidence: 99%

Widely-tunable synchronisation-free picosecond laser source for multimodal CARS, SHG, and two-photon microscopy

Xu¹,

Liang²,

Xu³

et al. 2021

Biomed. Opt. Express

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We demonstrate a continuous wave (CW) seeded synchronization-free optical parametric amplifier (OPA) pumped by a picosecond, 1 µm laser and show its performance when used as a simple yet powerful source for label-free coherent anti-Stokes Raman scattering (CARS), concurrent second harmonic generation (SHG), and two-photon fluorescence microscopy in an epi-detection geometry. The average power level of above 175 mW, spectral resolution of 8 cm −1 , and 2 ps pulse duration are well optimized for CARS microscopy in bio-science and bio-medical imaging systems. Our OPA is a much simpler setup than either the “gold-standard” laser and optical parametric oscillator (OPO) combination traditionally used for CARS imaging, or the more recently developed OPA systems pumped with femtosecond pulses [1]. Rapid and accurate tuning between resonances was achieved by changing the poled channels and temperature of the periodically-poled lithium niobate (PPLN) OPA crystal together with the OPA seed wavelength. The Pump-Stokes frequency detuning range fully covered the C-H stretching band used for the imaging of lipids. By enabling three multiphoton techniques using a compact, synchronization free laser source, our work paves the way for the translation of label-free multi-photon microscopy imaging from biomedical research to an imaging based diagnostic tool for use in the healthcare arena.

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Section: Resultsmentioning

confidence: 99%

Widely-tunable synchronisation-free picosecond laser source for multimodal CARS, SHG, and two-photon microscopy

Xu¹,

Liang²,

Xu³

et al. 2021

Biomed. Opt. Express

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show abstract

“…The morphology and distribution of ECM fibers are affected in several diseases, such as cancer, atherosclerosis, asthma, skin disorders [36][37][38][39], and the ability to accurately quantify and identify these changes bring unique perspectives related to disease mechanisms. In fibrosis, the ECM fibers are disorganized and present significant changes in thickness and orientation [40- 42], and access to such information has been bringing a new perspective in understanding for example, lung and cardiovascular diseases, such as asthma [10], and atherosclerosis [9].…”

Section: Discussionmentioning

confidence: 99%

Super resolution measurement of collagen fibers in biological samples: Validation of a commercial solution for multiphoton microscopy

et al. 2020

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Multiphoton microscopy is a powerful, non-invasive technique to image biological specimens. One current limitation of multiphoton microscopy is resolution as many of the biological molecules and structures investigated by research groups are similar in size or smaller than the diffraction limit. To date, the combination of multiphoton and super-resolution imaging has proved technically challenging for biology focused laboratories to implement. Here we validate that the commercial super-resolution Airyscan detector from ZEISS, which is based on image scanning microscopy, can be integrated under warranty with a pulsed multi-photon laser to enable multiphoton microscopy with super-resolution. We demonstrate its biological application in two different imaging modalities, second harmonic generation (SHG) and two-photon excited fluorescence (TPEF), to measure the fibre thicknesses of collagen and elastin molecules surpassing the diffraction limit by a factor of 1.7±0.3x and 1.4±0.3x respectively, in human heart and lung tissues, and 3-dimensional in vitro models. We show that enhanced resolution and signal-to-noise of SHG using the Airyscan compared to traditional GaAs detectors allows for automated and precise measurement of collagen fibres using texture analysis in biological tissues.

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“…Although some real-time video tracking systems has been proposed (23), their use is limited to the post-preparation phase and does not allow artery and vein discrimination. Recently, an in vivo vessel identification method by two-photon excitation elastin autofluorescence (TPEA) has been proposed (24). These authors showed a clear difference in the signal pattern of the vessel wall between artery and vein, but their method still did not satisfy the need for automatic vessel delineation and size measurement, and requires an expensive two-photon microscope.…”

Section: Discussionmentioning

confidence: 99%

Automated Detection and Diameter Estimation for Mouse Mesenteric Artery using Semantic Segmentation

Higaki

Mahmoud

Paradis

et al. 2020

Preprint

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Background: Pressurized myography is useful for the assessment of small artery structure and function, and widely used in the field of cardiovascular research. However, this procedure requires technical expertise for the sample preparation and effort to choose an appropriate size of artery. In this study we sought to develop an automatic artery-vein differentiation and size measurement system utilizing the U-Net-based machine learning algorithms. Methods and Results: We used 654 independent mesenteric artery images from 59 mice for the model training and validation. Our segmentation model yielded 0.744 ± 0.031 in IoU and 0.881 ± 0.016 in Dice coefficient with 5-fold cross validation. The vessel size and the lumen size calculated from the predicted vessel contours demonstrated a strong linear correlation with the manually determined vessel sizes (R = 0.722 ± 0.048, p<0.001 for vessel size and R = 0.908 ± 0.027, p<0.001 for lumen size). Lastly, we assessed the relation between the vessel size before and after dissection using pressurized myography system. We observed a strong positive correlation between the wall/lumen ratio before dissection and the lumen expansion ratio (R2 = 0.671, p<0.01). Using multivariate binary logistic regression, two models estimating whether the vessel met the size criteria (lumen size of 160 to 240 μm) were generated with area under the ROC curve of 0.761 for the upper limit and 0.747 for the lower limit. Conclusion: Our novel image analysis method with U-Net could streamline the experimental approach and may facilitate cardiovascular research.

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In vivo identification of arteries and veins using two‐photon excitation elastin autofluorescence

Cited by 15 publications

References 31 publications

Widely-tunable synchronisation-free picosecond laser source for multimodal CARS, SHG, and two-photon microscopy

Widely-tunable synchronisation-free picosecond laser source for multimodal CARS, SHG, and two-photon microscopy

Super resolution measurement of collagen fibers in biological samples: Validation of a commercial solution for multiphoton microscopy

Automated Detection and Diameter Estimation for Mouse Mesenteric Artery using Semantic Segmentation

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