Chinese Optics Letters, Volume. 23, Issue 7, 071701(2025)

Vascular permeability assessment using dual-wavelength photoacoustic microscopy with spectral unmixing

Yongyan Ren1,2, Kun Yu1,2, Qiansong Xia1,3, Honghui Li2、*, and Liming Nie1,2、**
Author Affiliations
  • 1School of Medicine, South China University of Technology, Guangzhou 510006, China
  • 2Medical Research Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China
  • 3School of Biomedical Engineering, Southern Medical University, Guangzhou 510515, China
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    Figures & Tables(6)
    Verification of spectral unmixing in phantom experiments. (a) Photograph of the in vitro phantom, displaying a plastic tube containing a mixture of blood and EB. (b) Absorption spectra of blood and EB. (c) Photoacoustic images of the phantom acquired at wavelengths of 532 and 610 nm. Scale bars = 400 µm. (d) Results of spectral unmixing, illustrating the separation of hemoglobin and EB signals in the mixtures. Scale bars = 300 µm. (e) Linear correlation between the photoacoustic signal intensity and EB concentration.
    Schematic diagram and performance testing of dual-wavelength PAM. (a) Schematic diagram of the dual-wavelength PAM system. CL, convex lens; PH, pinhole; DM, dichroic mirror; HWP, half-wave plate; FC, fiber coupler; SMF, single-mode fiber; NBF, narrow-bandpass filter; OL, objective lens; CorrL, correction lens; UT, ultrasound transducer; WT, water tank; AMP, amplifier; DAQ, data acquisition unit; PC, computer. (b) Photoacoustic image of a surgical blade, scale bar = 50 µm. (c) Scanning results of the sharp edges of the surgical blade. Selected B-scan images and their corresponding edge diffusion functions demonstrate lateral resolutions of 8.06 µm at 532 nm and 14.23 µm at 610 nm.
    Morphological alterations in the liver and kidney. Images of liver vasculature at 532 nm in (a) normal and (b) fibrotic mice (1-week model). PAM images of kidney vasculature at 532 nm in (c) normal and (d) UUO-treated fibrotic mice (1-week model). Scale bars = 200 µm for (a) and (b), and 100 µm for (c) and (d). (e) Quantitative analysis of vascular networks in the liver. (f) Quantitative analysis of vascular networks in the kidney. Statistical significance was assessed using a two-tailed unpaired Student’s t-test; *p p n = 3 per group).
    Time-lapse PAM imaging of liver EB extravasation. EB leakage in (a) normal and (b) fibrotic mice over a 15 min period post-injection. Scale bars = 100 µm. (c) Statistical comparison of EB dynamics. (d) Changes in hemoglobin concentration (CHb) due to fibrosis, analyzed through pixel comparisons. (e) EB extravasation at 15 min post-injection in control versus liver fibrosis groups. Statistical significance was assessed with a two-tailed unpaired Student’s t-test; **p p n = 3 per group).
    Time-lapse PAM imaging of kidney EB extravasation. EB leakage in the microvasculature of (a) normal and (b) UUO mice over 15 min post-injection. Scale bars = 100 µm. (c) Statistical comparison of EB dynamics. (d) Changes in CHb due to fibrosis analyzed by pixel comparisons. (e) EB extravasation at 15 min post-injection in control versus UUO mice. Statistical significance was assessed with a two-tailed unpaired Student’s t-test; **p n = 3 per group).
    Histopathological analysis of liver and kidney fibrosis. (a) Representative photomicrographs of liver tissues stained with hematoxylin-eosin and Masson’s trichrome. (b) Representative photomicrographs of kidney tissues stained with hematoxylin-eosin and Masson’s trichrome. Scale bars = 200 µm.
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    Yongyan Ren, Kun Yu, Qiansong Xia, Honghui Li, Liming Nie, "Vascular permeability assessment using dual-wavelength photoacoustic microscopy with spectral unmixing," Chin. Opt. Lett. 23, 071701 (2025)

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    Paper Information

    Category: Biophotonics

    Received: Jan. 25, 2025

    Accepted: Mar. 17, 2025

    Posted: Mar. 17, 2025

    Published Online: Jun. 20, 2025

    The Author Email: Honghui Li (lihonghui@gdph.org.cn), Liming Nie (limingnie@gmail.com)

    DOI:10.3788/COL202523.071701

    CSTR:32184.14.COL202523.071701

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