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谭天; 史天悦; 吴长锋; 彭洪尚

doi:10.37188/CO.2023-0070

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doi: 10.37188/CO.2023-0070

谭天 ^{1, 2,},
史天悦 ²,
吴长锋 ²,
彭洪尚 ^1,,

1.
中央民族大学理学院光子系统工程软件教育部工程研究中心, 北京海淀区 100081
2.
南方科技大学生物医学工程系, 广东省深圳市 518055

基金项目:国家自然科学基金（No. 62175266, No. 62235007, No. 22204070）；深圳市科技计划项目（No. KQTD20170810111314625, No. JCYJ20210324115807021, No. SGDX20211123114002003）；深圳湾实验室开放课题（No. SZBL2021080601002）；广东省先进生物材料重点实验室（No. 2022B1212010003）

详细信息

作者简介:
谭　天（1999—），男，湖北荆州人，硕士研究生，2020年于华中科技大学获得学士学位，现于中央民族大学攻读硕士学位，主要从事近红外激光扫描共聚焦系统及自适应光学方面的研究。E-mail：[email protected]

彭洪尚（1975—），男，山东临沂人，教授，博士研究生导师，2007年于北京交通大学光学获得理学博士，主要从事生物荧光纳米传感器和肿瘤光学医疗的研究。E-mail： [email protected]

中图分类号:O439
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365彩票官网网页版
- 网络出版日期: 2023-09-22

365彩票官网官方入口

TAN Tian ^{1, 2
,},
SHI Tian-yue ²,
WU Chang-feng ²,
PENG Hong-shang ^{1
,,}

1.
Engineering Research Center of Photonic Design Software Ministry of Education, College of Science, Minzu University of China, Beijing 100081, China
2.
Department of Biomedical Engineering, Southern University of Science and Technology, Shenzhen 518055, China

Funds:Supported by the National Natural Science Foundation of China (Grant No. 62175266, No. 62235007, No. 22204070); the Shenzhen Science and Technology Program (Grant No. KQTD20170810111314625, No. JCYJ20210324115807021, No. SGDX20211123114002003); the Shenzhen Bay Laboratory (No. SZBL2021080601002); and Guangdong Provincial Key Laboratory of Advanced Biomaterials (No. 2022B1212010003)

More Information

Corresponding author: [email protected]

摘要

摘要:
生物组织散射引起的光学像差限制了光学系统的成像性能。本文研究了基于间接波前整形的近红外二区荧光共聚焦成像技术。首先，合成制备了高效率近红外二区荧光探针，在该波段生物组织散射的降低有助于实现高对比度的活体组织成像。其次，研究了基于间接波前测量的自适应光学办法，将间接波前整形技术应用于激光扫描共聚焦显微系统中，以实现对于生物组织引起的光学像差的测量与补偿，获得生物组织的高信噪比成像。最后，对基于间接波前整形的近红外二区荧光共聚焦成像系统开展了相关实验。实验结果表明，本系统对空气平板、散射介质和小鼠颅骨等产生的像差具有良好的补偿效果，分别将最终信号强度提升为1.47、1.95和2.85倍，显著提升了最终的成像质量。
- 间接波前整形 /
- 近红外二区成像 /
- 共聚焦成像 /
- 活体实验
Abstract:
Optical aberrations caused by the scattering of biological tissues limit the imaging performance of optical systems. We have explored a fluorescence confocal imaging technique that utilizes wavefront shaping indirectly, operating in the near-infrared II range. First, we synthesized a highly efficient fluorescent probe in the near-infrared II range, where reducing the scattering of biological tissue can enable high-contrast biopsy imaging. Second, the study investigate the adaptive optical method, utilizing indirect wavefront measurement. The indirect wavefront shaping technology was applied to the laser scanning confocal system, enabling the measurement and compensation of optical aberrations caused by biological tissues, and obtaining imaging of biological tissues with a high signal-to-noise ratio. Finally, an indirect wavefront shaping-based near-infrared II fluorescence confocal imaging system was deployed and relavant experiments were conducted. The experimental outcomes reveal that the system effectively compensates for the aberrations induced by air plates, scattering media and mouse skull, and increases the final signal intensity to 1.47, 1.95 and 2.85 times, respectively. As a result, the final imaging quality is significantly enhanced.
- Indirect wavefront shaping /
- near-infrared-II imaging /
- confocal imaging /
- in vivo experiments

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图 1 半导体聚合物荧光探针的吸收与发射光谱

Figure 1. Absorption and emission spectra of the semiconductor polymer fluorescent probes

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图 2 基于间接波前测量的近红外激光共聚焦扫描显微系统

Figure 2. Near-infrared laser scanning confocal microscope based on indirect wavefront sensing

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图 3 控制SLM的反馈系统

Figure 3. Feedback system for controlling SLM

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图 4 系统像差的测试与校正（a）未进行校正时的样本成像；（b）经过系统像差校正后的样本成像图；（c）评价函数随迭代伦次的变化曲线；（d）GA算法计算得到的校正相位图；比例尺：（a）（b）全视场图中比例尺为200 μm，感兴趣区域局部放大图中比例尺为20 μm

Figure 4. Testing and correction of the system aberrations (a) Imaging of samples without correction; (b) Imaging of samples after systematic aberration correction; (c) Curve of the evaluation function as a function of iterative order; (d) The corrected phase diagram calculated by GA；Scale: 200 μm in the full field of view and 20 μm in the local magnification of the region of interest in (a) (b)

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图 5 电纺丝像差校正（a）引入一个30 μm的空气平板并仅进行系统像差校正的成像图；（b）在图（a）成像情况下，使用DASH算法进行像差校正；（c）在图（a）成像情况下，使用GA算法进行像差校正；（d）（a）（b）（c）中感兴趣区域的局部放大图白线标记处的荧光强度分布；（e）DASH算法得到的校正相位图；（f）GA算法得到的校正相位图；比例尺：（a）（b）（c）全视场图中比例尺为200 μm，感兴趣区域局部放大图中比例尺为20 μm

Figure 5. Aberration correction of electrospinning (a) Image with a 30 μm air plate and performing only system aberration correction; (b) In the case of imaging in figure (a), image with aberration correction by using DASH; (c) In the case of imaging in figure (a), image with aberration correction by using GA; (d) local magnification of the region of interest in (a) (b) (c) fluorescence intensity distribution at the white line marker; (e) the corrected phase map calculated by DASH; (f) The corrected phase diagram calculated by GA；Scale: 200 μm in the full field of view and 20 μm in the local magnification of the region of interest in (a) (b) (c)

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图 6 散射介质像差校正（a）直接成像图；（b）添加散射介质后的成像图；（c）添加介质后进行系统像差校正后的成像图；（d）添加介质后进行全像差校正后的成像图；（e）（a）-（d）中白线标记处的荧光强度分布；（f）GA算法计算得到的校正相位图；比例尺：（a）（b）（c）（d）全视场图中比例尺为200 μm，感兴趣区域局部放大图中比例尺为20 μm

Figure 6. Aberration correction of scattering medium (a) direct imaging (b) image after adding scattering medium; (c) image after system aberration correction; (d) image after total aberration correction; (e) distribution of fluorescence intensity at white line markers in (a)-(d); (f) the corrected phase map calculated by GA; Scale: 200 μm in the full field of view and 20 μm in the local magnification of the region of interest in (a) (b) (c) (d)

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图 7 活体小鼠颅内成像（a）仅进行系统像差校正的小鼠颅内血管成像图；（b）进行全像差校正后的小鼠颅内血管成像图；（c）（a）（b）中白线标记处的荧光强度分布；（d）评价函数随迭代伦次的变化曲线；比例尺：（a）（b）中比例尺为200 μm

Figure 7. Intracranial imaging of living mice (a) image of intracranial blood vessels in mice with only systematic aberration correction; (b) image of intracranial blood vessels in mice with full aberration correction; (c) the fluorescence intensity distribution at the white line mark in (a) and (b); (d) Curve of the evaluation function as a function of iterative order; scale: 200 μm in (a)(b).

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