1.中山大学中山医学院科研仪器管理中心,广东 广州 510080
2.中山大学热带病防治研究教育部重点实验室, 广东 广州 510080
3.广州实验室,广东 广州 510005
关苑君,硕士,助理实验师,研究方向:光学显微设备管理, E-mail: gyuanj@mail.sysu.edu.cn
收稿:2022-02-22,
纸质出版:2022-05-20
移动端阅览
关苑君,马显才.光学显微成像技术在液-液相分离研究中的应用[J].中山大学学报(医学科学版),2022,43(03):504-510.
GUAN Yuan-jun,MA Xian-cai.Application of Optical Microscopic Imaging Technology in Liquid-Liquid Separation Research[J].Journal of Sun Yat-sen University(Medical Sciences),2022,43(03):504-510.
关苑君,马显才.光学显微成像技术在液-液相分离研究中的应用[J].中山大学学报(医学科学版),2022,43(03):504-510. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2022.0319.
GUAN Yuan-jun,MA Xian-cai.Application of Optical Microscopic Imaging Technology in Liquid-Liquid Separation Research[J].Journal of Sun Yat-sen University(Medical Sciences),2022,43(03):504-510. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2022.0319.
近十年来,科学家们发现液-液相分离在许多生理过程中扮演重要角色,发挥着许多生物学功能。一旦发生异常,将会导致多种疾病产生。在生物学研究中,光学显微成像技术是液-液相分离主要的研究工具之一。本文阐述了几个重要的光学显微成像技术在液-液相分离的研究中的应用和对比,以观察相分离的存在和形成等过程。详细地阐述了运用共聚焦显微技术中的荧光漂白后恢复技术(FRAP)检测其扩散和物质交换的特性,并总结了应用该技术时的成像条件、注意事项和参数应用,为科学研究者提供技术选择的依据。
In the past decade, scientists have found that liquid-liquid separation plays an important role in many physiological processes and plays many biological functions. Its abnormal change will lead to a variety of diseases. In the study of liquid-liquid separation in biology, optical microscopy is the main research tool. This paper describes the application and comparison of several important optical microscopic imaging technologies in the study of liquid-liquid phase separation, to explain the existence and formation process of phase separation, obtain the mobility and diffusion data. The characteristics of diffusion and material exchange detected by fluorescence recovery after photobleaching (FRAP) in confocal microscopy are described in detail, and the imaging conditions, precautions and parameters are summarized. That might provide the basis for scientific researchers to choose the technology.
Hyman AA , Weber CA , Jülicher F . Liquid-liquid phase separation in biology [J]. Annu Rev Cell Dev Biol , 2014 , 30 : 39 - 58 .
吴荣波 , 李丕龙 . 液-液相分离与生物分子凝集体 [J]. 科学通报 , 2019 , 64 ( 22 ): 2285 - 2291 .
Wu RB , Li PL . Liquid-liquid phase separation and biomolecular condensates [J]. Chin Sci Bull , 2019 , 64 ( 22 ): 2285 - 2291 .
Patel A , Lee H , Jawerth L , et al . A liquid-to-solid phase transition of the ALS protein FUS accelerated by disease mutation [J]. Cell , 2015 , 162 ( 5 ): 1066 - 1077 .
Molliex A , Temirov J , Lee J , et al . Phase separation by low complexity domains promotes stress granule assembly and drives pathological fibrillization [J]. Cell , 2015 , 163 ( 1 ): 123 - 133 .
Wegmann S , Eftekharzadeh B , Tepper K , et al . Tau protein liquid-liquid phase separation can initiate tau aggregation [J]. EMBO J , 2018 , 37 ( 7 ): e98049 .
Boulay G , Sandoval GJ , Riggi N , et al . Cancer-specific retargeting of BAF complexes by a prion-like domain [J]. Cell , 2017 , 171 ( 1 ): 163 - 178.e19 .
Franzmann TM , Jahnel M , Pozniakovsky A , et al . Phase separation of a yeast prion protein promotes cellular fitness [J]. Science , 2018 , 359 ( 6371 ): eaao5654 .
Banani S , Lee H , Hyman A , et al . Biomolecular condensates: organizers of cellular biochemistry [J]. Nat Rev Mol Cell Biol , 2017 , 18 ( 5 ): 285 - 298 .
Larson A , Narlikar G . The role of phase separation in heterochromatin formation, function, and regulation [J]. Biochemistry , 2018 , 57 ( 17 ): 2540 - 2548 .
Sabari B , Dall’agnese A , Boija A , et al . Coactivator condensation at super-enhancers links phase separation and gene control [J]. Science , 2018 , 361 ( 6400 ): eaar3958 .
Chong S , Dugast-darzacq C , Liu Z , et al . Imaging dynamic and selective low-complexity domain interactions that control gene transcription [J]. Science , 2018 , 361 ( 6400 ): eaar2555 .
Shin Y , Chang Y , Lee D , et al . Liquid nuclear condensates mechanically sense and restructure the genome [J]. Cell , 2018 , 175 ( 6 ): 1481 - 1491.e13 .
Widom B . Note on the interfacial tension of phase-separated polymer solutions [J]. Journal of Statistical Physics , 1988 , 52 : 1343 – 1351 .
Alberti S , Gladfelter A , Mittag T . Considerations and challenges in studying liquid-liquid phase separation and biomolecular condensates [J]. Cell , 2019 , 176 ( 3 ): 419 - 434 .
Shin Y , Berry J , Pannucci N , et al . Spatiotemporal control of intracellular phase transitions using light-activated optoDroplets [J]. Cell , 2017 , 168 ( 1-2 ): 159 - 171 .e14.
Zeng M , Shang Y , Araki Y , et al . Phase transition in postsynaptic densities underlies formation of synaptic complexes and synaptic plasticity [J]. Cell , 2016 , 166 ( 5 ): 1163 - 1175.e12 .
刘向阳 . 全内反射亚表面损伤检测中加入DIC显微术的研究和应用 [D]. 南京 : 南京理工大学 , 2017 .
Liu XY . Design and application of DIC microscopy in detection of total internal reflected sub-surface damage [D]. Nanjing : Nanjing University of Science and Technology , 2017 .
张祥翔 . 现代显微成像技术综述 [J]. 光学仪器 , 2015 , 37 ( 6 ): 550 - 560 .
Zhang XX . Imaging technology in modern microsystems [J]. Opt Inst , 2015 , 37 ( 6 ): 550 - 560 .
Li P , Banjade S , Cheng H , et al . Phase transitions in the assembly of multivalent signaling proteins [J]. Nature , 2012 , 483 ( 7389 ): 336 - 340 .
Plys A , Davis C , Kim J , et al . Phase separation of polycomb-repressive complex 1 is governed by a charged disordered region of CBX2 [J]. Genes Dev , 2019 , 33 ( 13-14 ): 799 - 813 .
Zhou M , Li W , Li J , et al . Phase-separated condensate-aided enrichment of biomolecular interactions for high-throughput drug screening in test tubes [J]. J Biol Chem , 2020 , 295 ( 33 ): 11420 - 11434 .
Fu Y , Zhuang X . m6A-binding YTHDF proteins promote stress granule formation [J]. Nat Chem Biol , 2020 , 16 ( 9 ): 955 - 963 .
Zeng W , Lu C , Shi Y , et al . Initiation of stress granule assembly by rapid clustering of IGF2BP proteins upon osmotic shock [J]. Biochim Biophys Acta Mol Cell Res , 2020 , 1867 ( 10 ): 118795 .
Agote-aran A , Schmucker S , Jerabkova K , et al . Spatial control of nucleoporin condensation by fragile X-related proteins [J]. EMBO J , 2020 , e104467 .
Ma X , Chen T , Peng Z , et al . Histone chaperone CAF-1 promotes HIV-1 latency by leading the formation of phase-separated suppressive nuclear bodies [J]. EMBO J , 2021 , e106632 .
张志毅 , 周涛 , 巩伟丽 , 等 . 荧光漂白后恢复技术及其在活细胞分子机制研究中的应用 [J]. 生物技术通讯 , 2008 , ( 4 ): 635 - 637 .
Zhang ZY , Zhou T , Gong WL , et al . Fluorescence recovery after photobleaching and its applications in the study of cellular molecular mechanics [J]. Lett Biotechnol , 2008 , ( 4 ): 635 - 637 .
Mcswiggen D , Mir M , Darzacq X , et al . Evaluating phase separation in live cells: diagnosis, caveats, and functional consequences [J]. Genes Dev , 2019 , 33 ( 23-24 ): 1619 - 1634 .
Taylor N , Wei M , Stone H , et al . Quantifying dynamics in phase-separated condensates using fluorescence recovery after photobleaching [J]. Biophys J , 2019 , 117 ( 7 ): 1285 - 1300 .
Lele T , Ingber D . A mathematical model to determine molecular kinetic rate constants under non-steady state conditions using fluorescence recovery after photobleaching (FRAP) [J]. Biophys Chem , 2006 , 120 ( 1 ): 32 - 35 .
Weidemann T . Application of fluorescence correlation spectroscopy (FCS) to measure the dynamics of fluorescent proteins in living cells [J]. Methods Mol Biol , 2014 , 1076 : 539 - 55 .
董辰博 , 李福才 , 董朝青 , 等 . 激光共聚焦扫描成像-荧光相关光谱系统的研究及其应用 [J]. 分析科学学报 , 2020 , 36 ( 5 ): 695 - 700 .
Dong CB , Li FC , Dong CQ , et al . The developing of laser confocal scanning imaging-fluorescence correlation spectroscopy system and its application [J]. J Anal Sci , 2020 , 36 ( 5 ): 695 - 700 .
于淼 , 高建 . 荧光漂白恢复、荧光共振能量转移和荧光相关光谱检测的技术特点 [J]. 中国医学装备 , 2009 , 6 ( 6 ): 8 - 9 .
Yu M , Gao J . Characteristics of application and technology on FRAP, FRET and FCS [J]. Chin Med Equip , 2009 , 6 ( 6 ): 8 - 9 .
Wei M , Elbaum-garfinkle S , Holehouse A , et al . Phase behavior of disordered proteins underlying low density and high permeability of liquid organelles [J]. Nat Chem , 2017 , 9 ( 11 ): 1118 - 1125 .
许乔 . 基于拉曼光谱及偏光成像技术的阿尔兹海默症相关物质的分析研究 [D]. 上海 : 华东师范大学 , 2018 .
Xu Q . Analysis of relative species in the Alzheimer’s diseases based on Raman spectroscopy and polarized light microscopy [D]. Shanghai : East China Normal University , 2018 .
Shin Y , Chang Y , Lee D , et al . Liquid nuclear condensates mechanically sense and restructure the genome [J]. Cell , 2019 , 176 ( 6 ): 1518 .
0
浏览量
408
下载量
2
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621
