现代仪器与医疗

活体显微成像技术研究进展

PDF下载

俞方菲 ¹^,² , 卢志 ¹^,³ , 江劭玮 ² , 杨懿 ¹^,⁴^,⁵^,⁶^,^*

现代仪器与医疗 | 生物应用 2025,31(4): 73-80

收起

现代仪器与医疗 |生物应用 2025 , 31 (4) : 73 -80

活体显微成像技术研究进展

全屏

俞方菲¹^,², 卢志¹^,³, 江劭玮², 杨懿¹^,⁴^,⁵^,⁶^,^*

作者信息

^1.浙江荷湖科技有限公司，杭州　311113

^2.杭州电子科技大学通信工程学院，杭州　310018

^3.清华大学自动化系，北京　100084

^4.北京航空航天大学可靠性与系统工程学院，北京　100191

^5.鹏城实验室，深圳　518000

^6.北京航空航天大学杭州创新研究院，杭州　310052

通讯作者:

杨懿，女，博士研究生，研究员，研究方向：人工智能算法、复杂系统多源数据融合、系统可靠性预测以及控制理论与应用等

作者简介:

俞方菲，女，博士研究生，研究方向：肿瘤免疫

Research progress of intravital imaging

Fangfei Yu¹^,², Zhi Lu¹^,³, Shaowei Jiang², Yi Yang¹^,⁴^,⁵^,⁶^,^*

Affiliations

^1.Zhejiang Hehu Technology Co., Ltd., Hangzhou 311113, China

^2.School of Communication Engineering, Hangzhou Dianzi University, Hangzhou 310018, China

^3.Department of Automation, Tsinghua University, Beijing 100084, China

^4.School of Reliability and Systems Engineering, Beihang University, Beijing 100191, China

^5.Peng Cheng Laboratory, Shenzhen 518000, China

^6.Hangzhou Innovation Institute of Beihang University, Hangzhou 310052, China

出版时间: 2025-08-10 doi: 10.11876/mimt202504008

文章导航

摘要

收起

活体显微成像技术作为生命科学研究的核心工具，能够在自然生理环境中实时追踪细胞及分子动态，为解析复杂生物过程提供了前所未有的视角。本文系统阐述共聚焦显微镜、多光子显微镜、光片显微镜和扫描光场显微镜的成像原理、技术特性及局限性，结合肿瘤学、免疫学、神经科学和发育生物学等领域的典型应用案例，横向对比不同技术的适用场景，并基于当前技术瓶颈展望未来发展方向，以期为相关研究提供技术选择参考。

关键词

活体显微成像 / 共聚焦显微镜 / 多光子显微镜 / 光片显微镜 / 扫描光场显微镜

Abstract

收起

As a pivotal tool in life science research, intravital microscopy facilitates real-time monitoring of cellular and molecular dynamics within their native physiological milieu. This review delves into the imaging mechanisms, strengths, and constraints of confocal microscopy, multi-photon microscopy, light-sheet microscopy, and scanning light-field microscopy. By analyzing representative application cases across disciplines such as oncology, immunology, neuroscience, and developmental biology, it offers a comparative overview of the ideal usage scenarios for these technologies. Furthermore, it addresses current technical hurdles and speculates on future advancements in the field. To provide technical reference for related research.

Key words

Intravital imaging / Confocal microscopy / Multi-photon microscopy / Light-sheet microscopy / Scanning light-field microscopy

引用本文

俞方菲, 卢志, 江劭玮, 杨懿. 活体显微成像技术研究进展. 现代仪器与医疗, 2025 , 31 (4) : 73 -80 . DOI: 10.11876/mimt202504008

Fangfei Yu, Zhi Lu, Shaowei Jiang, Yi Yang. Research progress of intravital imaging[J]. Modern Instruments & Medical Treatment, 2025 , 31 (4) : 73 -80 . DOI: 10.11876/mimt202504008

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

[1]

Su J, Song Y, Zhu Z, et al. Cell–cell communication: new insights and clinical implications[J]. Signal Transduction and Targeted Therapy, 2024, 9: 196.

[2]

Zhu Y, Chen J, Chen C, et al. Deciphering mechanical cues in the microenvironment: from non-malignant settings to tumor progression[J]. Biomarker Research, 2025, 13(1): 11.

[3]

Mempel T R, Lill J K, Altenburger L M. How chemokines organize the tumour microenvironment[J]. Nature Reviews Cancer, 2024, 24(1): 28-50.

[4]

Pittet M J, Weissleder R. Intravital imaging[J]. Cell, 2011, 147(5): 983-991.

[5]

Stephens D J, Allan V J. Light microscopy techniques for live cell imaging[J]. Science, 2003, 300(5616): 82-86.

[6]

Entenberg D, Oktay M H, Condeelis J S. Intravital imaging to study cancer progression and metastasis[J]. Nature Reviews Cancer, 2023, 23: 25-42.

[7]

Peng X, Wang Y, Zhang J, et al. Intravital imaging of the functions of immune cells in the tumor microenvironment during immunotherapy[J]. Frontiers in Immunology, 2023, 14: 1288273.

[8]

Honda M, Kadohisa M, Yoshii D, et al. Intravital imaging of immune responses in intestinal inflammation[J]. Inflammation and Regeneration, 2023, 43: 9.

[9]

White J G, Amos W B, Fordham M. An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy[J]. The Journal of cell biology, 1987, 105(1): 41-48.

[10]

Nwaneshiudu A, Kuschal C, Sakamoto F H, et al. Introduction to confocal microscopy[J]. Journal of Investigative Dermatology, 2012, 132(12): 1-5.

[11]

Wan Y, McDole K, Keller P J. Light-sheet microscopy and its potential for understanding developmental processes[J]. Annual Review of Cell and Developmental Biology, 2019, 35: 655-681.

[12]

Ding Y, Ma J, Langenbacher A D, et al. Multiscale light-sheet for rapid imaging of cardiopulmonary system[J]. JCI Insight, 2018, 3(16): e121396.

[13]

Chow D J X, Schartner E P, Corsetti S, et al. Quantifying DNA damage following light sheet and confocal imaging of the mammalian embryo[J]. Scientific Reports, 2024, 14: 20760.

[14]

Denk W, Strickler J H, Webb W W. Two-photon laser scanning fluorescence microscopy[J]. Science, 1990, 248(4951): 73-76.

[15]

Campagnola P J, Loew L M. Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms[J]. Nature Biotechnology, 2003, 21(11): 1356-1360.

[16]

James D S, Campagnola P J. Recent advancements in optical harmonic generation microscopy: applications and perspectives[J]. BME Frontiers, 2021: 3973857.

[17]

Hennessy N, Simms C. Skeletal muscle extracellular matrix structure under applied deformation observed using second harmonic generation microscopy[J]. Acta Biomaterialia, 2023, 172: 135-146.

[18]

Seeger M, Karlas A, Soliman D, et al. Multimodal optoacoustic and multiphoton microscopy of human carotid atheroma[J]. Photoacoustics, 2016, 4(3): 102-111.

[19]

Lu Z, Liu Y, Jin M, et al. Virtual-scanning light-field microscopy for robust snapshot high-resolution volumetric imaging[J]. Nature Methods, 2023, 20(5): 735-746.

[20]

Lu Z, Jin M, Chen S, et al. Physics-driven self-supervised learning for fast high-resolution robust 3D reconstruction of light-field microscopy[J]. Nature Methods, 2025, 22(7): 1545–1555.

[21]

Wu J, Lu Z, Jiang D, et al. Iterative tomography with digital adaptive optics permits hour-long intravital observation of 3D subcellular dynamics at millisecond scale[J]. Cell, 2021, 184(12): 3318-3332.

[22]

Lu Z, Cai Y, Nie Y, et al. A practical guide to scanning light-field microscopy with digital adaptive optics[J]. Nature Protocols, 2022, 17(9): 1953-1979.

[23]

Lu Z, Zuo S, Shi M, et al. Long-term intravital subcellular imaging with confocal scanning light-field microscopy[J]. Nature Biotechnology, 2025, 43(4): 569-580.

[24]

Burkard P, Schonhart C, Vögtle T, et al. A key role for platelet GPVI in neutrophil recruitment, migration, and NETosis in the early stages of acute lung injury[J]. Blood, 2023, 142(17): 1463-1477.

[25]

Park I, Kim S, Um Y W, et al. Intravital imaging of peritubular microcirculation impairment in cisplatin-induced acute kidney injury[J]. JCI Insight, 2025, 10(9): e178689.

[26]

Lohela M, Casbon A J, Olow A, et al. Intravital imaging reveals distinct responses of depleting dynamic tumor-associated macrophage and dendritic cell subpopulations[J]. Proceedings of the National Academy of Sciences, 2014, 111(47): E5086-E5095.

[27]

Lee J, Balzraine B, Schweizer A, et al. Neutrophil CRACR2A promotes neutrophil recruitment in sterile inflammation and ischemic stroke[J]. Circulation, 2025, 151(10): 696-715.

[28]

Almstedt E, Rosén E, Gloger M, et al. Real-time evaluation of glioblastoma growth in patient-specific zebrafish xenografts[J]. Neuro-Oncology, 2022, 24(5): 726-738.

[29]

Eum J, Kwak J, Kim H J, et al. 3D visualization of developmental toxicity of 2,4,6-trinitrotoluene in zebrafish embryogenesis using light-sheet microscopy[J]. International Journal of Molecular Sciences, 2016, 17(11): 1925.

[30]

Djenoune L, Mahamdeh M, Truong T V, et al. Cilia function as calcium-mediated mechanosensors that instruct left-right asymmetry[J]. Science, 2023, 379(6627): 71-78.

[31]

Bornhorst D, Hejjaji A V, Steuter L, et al. The heart is a resident tissue for hematopoietic stem and progenitor cells in zebrafish[J]. Nature Communications, 2024, 15(1): 7589.

[32]

Gaertner F, Ishikawa-Ankerhold H, Stutte S, et al. Plasmacytoid dendritic cells control homeostasis of megakaryopoiesis[J]. Nature, 2024, 631(8021): 645-653.

[33]

Shen Y, Hu M, Li W, et al. Delivery of DNA octahedra enhanced by focused ultrasound with microbubbles for glioma therapy[J]. Journal of Controlled Release, 2022, 350: 158-174.

[34]

Junankar S, Shay G, Jurczyluk J, et al. Real-time intravital imaging establishes tumor-associated macrophages as the extraskeletal target of bisphosphonate action in cancer[J]. Cancer Discovery, 2015, 5(1): 35-42.

[35]

Lämmermann T, Afonso P V, Angermann B R, et al. Neutrophil swarms require LTB4 and integrins at sites of cell death in vivo[J]. Nature, 2013, 498(7454): 371-375.

[36]

Bhattacharyya N D, Kyaw W, McDonald M M, et al. Minimally invasive longitudinal intravital imaging of cellular dynamics in intact long bone[J]. Nature Protocols, 2023, 18(12): 3856-3880.

[37]

Zhang Y, Wang M, Zhu Q, et al. Long-term mesoscale imaging of 3D intercellular dynamics across a mammalian organ[J]. Cell, 2024, 187(21): 6104-6122.

[38]

Li Y, Liu T M. Discovering macrophage functions using in vivo optical imaging techniques[J]. Frontiers in Immunology, 2018, 9: 502.

[39]

Xiao Y, Faucherre A, Pola-Morell L, et al. High-resolution live imaging reveals axon-glia interactions during peripheral nerve injury and repair in zebrafish[J]. Disease Models & Mechanisms, 2015, 8(6): 553-564.

[40]

Nguyen D X, Bos P D, Massagué J. Metastasis: from dissemination to organ-specific colonization[J]. Nature Reviews Cancer, 2009, 9: 274-284.

[41]

Fuhrmann F, Nebeling F C, Musacchio F, et al. Three-photon in vivo imaging of neurons and glia in the medial prefrontal cortex with sub-cellular resolution[J]. Communications Biology, 2025, 8: 795.

[42]

Zhang Y, Yuan L, Zhu Q, et al. A miniaturized mesoscope for the large-scale single-neuron-resolved imaging of neuronal activity in freely behaving mice[J]. Nature Biomedical Engineering, 2024, 8(6): 754-774.

[43]

Ki S, Kwon S H, Eum J, et al. 3D light-sheet assay assessing novel valproate-associated cardiotoxicity and folic acid relief in zebrafish embryogenesis[J]. Chemosphere, 2019, 227: 551-560.

[44]

Liu T L, Upadhyayula S, Milkie D E, et al. Observing the cell in its native state: imaging subcellular dynamics in multicellular organisms[J]. Science, 2018, 360(6386): eaaq1392.

[45]

Matchett K P, Wilson-Kanamori J R, Portman J R, et al. Multimodal decoding of human liver regeneration[J]. Nature, 2024, 630(8015): 158-165.

[46]

Chen S, Lu Z, Zhao Y, et al. Myeloid-mas signaling modulates pathogenic crosstalk among MYC+ CD63+ endothelial cells, MMP12+ macrophages, and monocytes in acetaminophen-induced liver injury[J]. Advanced Science, 2024, 11(16): e2306066.

[47]

Mencattini A, Di Giuseppe D, Comes M C, et al. Discovering the hidden messages within cell trajectories using a deep learning approach for in vitro evaluation of cancer drug treatments[J]. Scientific Reports, 2020, 10(1): 7653.

[48]

Ravindran S. Five ways deep learning has transformed image analysis[J]. Nature, 2022, 609(7928): 864-866.

[49]

Karim M M, Sun R, Khajavi B, et al. Multimodal optical coherence tomography and two-photon light sheet fluorescence microscopy for embryo imaging[J]. Journal of Biomedical Optics, 2025, 30(6): 060501.

[50]

Harder A, Nagarajan B, Odermatt B, et al. Automatic detector synchronization for long-term imaging using confocal light-sheet microscopy[J]. Microscopy Research and Technique, 2023, 86(2): 125-136.

[51]

Saitou T, Imamura T. Extended depth of focus two-photon light-sheet microscopy for in vivo fluorescence imaging of large multicellular organisms at cellular resolution[J]. International Journal of Molecular Sciences, 2023, 24(12): 10186.

[52]

Zhao Z, Zhou Y, Liu B, et al. Two-photon synthetic aperture microscopy for minimally invasive fast 3D imaging of native subcellular behaviors in deep tissue[J]. Cell, 2023, 186(11): 2475-2491.

[53]

MacRae C A, Peterson R T. Zebrafish as a mainstream model for in vivo systems pharmacology and toxicology[J]. Annual Review of Pharmacology and Toxicology, 2023, 63: 43-64.

[54]

Zhou L, Huang J, Li C, et al. Organoids and organs-onchips: recent advances, applications in drug development, and regulatory challenges[J]. Med, 2025, 6(4): 100667.

[55]

Du X, Jia H, Chang Y, et al. Progress of organoid platform in cardiovascular research[J]. Bioactive Materials, 2024, 40: 88-103.

[56]

Wakefield L, Agarwal S, Tanner K. Preclinical models for drug discovery for metastatic disease[J]. Cell, 2023, 186(8): 1792-1813.

[57]

Ricard C, Hazart D, Delhomme B, et al. Fast tissue clearing and volume imaging techniques for anatomy[J/OL]. Microscopy Research and Technique, 2025[2025-06-09]. https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/jemt.70013.

[58]

Zhao X, Pellow C, Goertz D E. Intravital imaging and cavitation monitoring of antivascular ultrasound in tumor microvasculature[J]. Theranostics, 2023, 13(1): 250-266.

[59]

Ahn S, Yoon J Y, Kim P. Intravital imaging of cardiac tissue utilizing tissue-stabilized heart window chamber in live animal model[J]. European Heart Journal Imaging Methods and Practice, 2024, 2(3): qyae062.

[60]

Entenberg D, Voiculescu S, Guo P, et al. A permanent window for the murine lung enables high-resolution imaging of cancer metastasis[J]. Nature Methods, 2018, 15(1): 73-80.

[61]

Zhu X, Huang Q, Jiang L, et al. Longitudinal intravital imaging of mouse placenta[J]. Science Advances, 2024, 10(12): eadk1278.

[62]

Gao X, Xiong Y, Ma H, et al. Visualizing bulk autophagy in vivo by tagging endogenous LC3B[J]. Autophagy, 2025, 21(7): 1591-1607.

[63]

Fan X, Zhou Y, Bai W, et al. Intravital imaging of translocated bacteria via fluorogenic labeling of gut microbiota in situ[J]. Proc Natl Acad Sci USA, 2025, 122(13): e2415845122.

[64]

Zhanghao K, Li M, Chen X, et al. Fast segmentation and multiplexing imaging of organelles in live cells[J]. Nature Communications, 2025, 16: 2769.

[65]

Alieva M, Wezenaar A K L, Wehrens E J, et al. Bridging live-cell imaging and next-generation cancer treatment[J]. Nature Reviews Cancer, 2023, 23(11): 731-745.

[66]

Jorch S K, Deppermann C. Intravital imaging allows organ-specific insights into immune functions[J]. Frontiers in Cell and Developmental Biology, 2021, 9: 623906.

[67]

Wang H, Li L, Li Y, et al. Intravital imaging of interactions between iNKT and kupffer cells to clear free lipids during steatohepatitis[J]. Theranostics, 2021, 11(5): 2149-2169.

[68]

Castanheira F V S, Nguyen R, Willson M, et al. Intravital imaging of three different microvascular beds in SARS-CoV-2–infected mice[J]. Blood Advances, 2023, 7(15): 4170-4181.

[69]

Stern M A, Dingledine R, Gross R E, et al. Epilepsy insights revealed by intravital functional optical imaging[J]. Frontiers in Neurology, 2024, 15: 1465232.

[70]

Giampetraglia M, Weigelin B. Recent advances in intravital microscopy for preclinical research[J]. Current Opinion in Chemical Biology, 2021, 63: 200-208.

[71]

Yang M, Hou S, Chen Y, et al. Emerging insights into intravital imaging, unraveling its role in cancer immunotherapy[J]. Cancer Immunology, Immunotherapy, 2025, 74: 100.

[72]

Ko J, Hyung S, Cheong S, et al. Revealing the clinical potential of high-resolution organoids[J]. Advanced Drug Delivery Reviews, 2024, 207: 115202.

[73]

König K, Breunig H G, Batista A, et al. Translation of two-photon microscopy to the clinic: multimodal multiphoton CARS tomography of in vivo human skin[J]. Journal of Biomedical Optics, 2020, 25(1): 014515.

[74]

Lentsch G, Baugh E G, Lee B, et al. Research techniques made simple: emerging imaging technologies for noninvasive optical biopsy of human skin[J]. The Journal of investigative dermatology, 2022, 142(5): 1243-1252.

2025年第31卷第4期

PDF下载

引用本文

BibTeX

文章信息

doi: 10.11876/mimt202504008

首发时间：2026-10-23
出版时间：2025-08-10

补充材料

相关文章

文章信息

作者

出版历史

基金

作者信息

^1.浙江荷湖科技有限公司，杭州　311113

^2.杭州电子科技大学通信工程学院，杭州　310018

^3.清华大学自动化系，北京　100084

^4.北京航空航天大学可靠性与系统工程学院，北京　100191

^5.鹏城实验室，深圳　518000

^6.北京航空航天大学杭州创新研究院，杭州　310052

通讯作者:

杨懿，女，博士研究生，研究员，研究方向：人工智能算法、复杂系统多源数据融合、系统可靠性预测以及控制理论与应用等

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/xdyqyyl/CN/10.11876/mimt202504008

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT