直播预告 | 相干拉曼散射成像线上研讨会

2021-10-15 17:06:08, 徕卡显微系统 徕卡显微系统(上海)贸易有限公司


直播主题

相干拉曼散射成像线上研讨会

直播时间

10月20日

报名方式

长按识别二维码

预约报名


直播议程

时间

分享主题

分享嘉宾

14:00-14:30

受激拉曼显微术用于快速无标记组织病理学检测

季敏标

14:30-15:00

受激拉曼激发荧光光谱学与显微成像

熊汗青

15:00-15:30

超快超分辨受激拉曼散射显微镜用于特定代谢分子与蛋白成像

王平

15:30-16:00

Leica STELLARIS 8 CRS相干拉曼散射系统介绍

蒋忠祥

16:00-16:20

样机演示

蒋忠祥,

Volker Schweikh

16:20-16:30

讨论环节

在利用显微镜进行微观世界观察的时候,我们往往需要利用一些方法来提高反差,从而识别样品的结构,例如图一左图的微分干涉(DIC),中图的相差(PH)和右图的荧光成像(FL)。前两种方法无需对样品做额外的标记即可成像,方便快捷,但是能够区分的结构有限;荧光成像的方法能够观察到一些特异性的结构,但是需要对特异性结构进行一些标记,这个过程需要比较长的耗时,且会对生物体本身产生一些影响。所有就会有些科学家在探索一些免标记成像的方法。

图一 提高图像反差

比如利用生物体的自发荧光,SHG和THG等荧光信号(如图二),另一种免标记成像则是拉曼散射(如图三)。我们通常讲的拉曼散射指自发拉曼散射过程,信号非常弱(比荧光低 8-10 个数量级),无法用于快速成像。相干拉曼是增强拉曼的手段之一,利用短脉冲激光的非线性效应实现增强,可以有效的用于生物样品的三维(视频一)和活体成像(视频二)。近几年采用该方法发表的文章也越来越多,广泛的应用在癌症研究,病理学研究,神经生物学,药学,代谢,免疫,植物,材料等领域(见附件文献列表),此外,还有科学家利用拉曼光谱的窄带特性,研制了一系列共振探针,从而可以轻松实现多达十色标记的生物学成像。

图二 5日龄斑马鱼胚胎尾部区域。灰色:920nm SHG信号,红色:自发荧光,青色:1260nm THG信号


图三 斑马鱼眼部无标记成像。绿色:脂类 (SRS, 2850 cm-1),红色:蛋白质(SRS, 2930 cm-1)


图四 Hela细胞十色标记成像,每张图片选择两通道叠加。PM (Carbow2141), ER (Carbow2226), Golgi (BODIPY TR), Mito (Carbow2062), LD (Carbow2202), Lyso (Carbow2086), nucleus (NucBlue), tubulin (SiR650), actin (GFP) and FM 4-64。

文献出处:Supermultiplexed optical imaging and barcoding with engineered polyynes. Hu F, Zeng C, Long R, Miao Y, Wei L, Xu Q, Min W. Nat Methods. 2018 Mar;15(3):194-200. doi: 10.1038/nmeth.4578.

视频一:小鼠脑部三维切片。橙色:脂类(SRS, 2850cm-1);青色:Thy-1 YFP-16荧光信号。


视频二:小肠类器官。显示脂类的动态运动。


相干拉曼散射成像对于生物学领域的研究者来说,还是比较陌生的领域,为了向广大研究者介绍这项快速发展的新技术,徕卡显微系统特联合丁香园举办此次“相干拉曼散射成像线上研讨会”,邀请到相干拉曼散射领域研究的先驱者和专家们向大家介绍这项技术的原理,特性和在多个领域的应用情况。


欢迎有兴趣的老师们注册参加,如果需要了解一些背景知识,也可以参考之前的一些文章:

大咖讲堂 | 相干拉曼散射显微术Ⅰ

► 点击阅读

大咖讲堂 | 相干拉曼散射显微术 Ⅱ

► 点击阅读


*部分研究结果清单,更详细信息可参考https://www.leica-microsystems.com/science-lab/cars-publication-list/

参考文献:

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病理学

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High-Speed Coherent Raman Fingerprint Imaging of Biological Tissues.
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神经生物学

Lipid-droplet-accumulating microglia represent a dysfunctional and proinflammatory state in the aging brain
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Nat Neurosci. 2020 Feb;23(2):194-208. doi: 10.1038/s41593-019-0566-1. Epub 2020 Jan 20.

Lewy pathology in Parkinson''s disease consists of crowded organelles and lipid membranes
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Nat Neurosci. 2019 Jul;22(7):1099-1109. doi: 10.1038/s41593-019-0423-2. Epub 2019 Jun 24.

Label-free imaging of amyloid plaques in Alzheimer’s disease with stimulated Raman scattering microscopy,
M. Ji, M.Arbel, L. Zhang, C.W. Freudiger, S.S. Hou, D. Lin, X. Yang, B.J. Bacskai and X.S. Xie.
Sci Adv. 2018 Nov; 4(11): eaat7715. Published online 2018 Nov 16. doi: 10.1126/sciadv.aat7715

Label-free Imaging of Neurotransmitter Acetylcholine at Neuromuscular Junctions with Stimulated Raman Scattering.
D. Fu, W. Yang, X.S. Xie
J Am Chem Soc. 2017 Jan 18;139(2):583-586. doi: 10.1021/jacs.6b10727. Epub 2016 Dec 30.

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药学

Raman Imaging of Nanocarriers for Drug Delivery.
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Detecting and Quantifying Microscale Chemical Reactions in Pharmaceutical Tablets by Stimulated Raman Scattering Microscopy.
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Imaging the Intracellular Distribution of Tyrosine Kinase Inhibitors in Living Cells with Quantitative Hyperspectral Stimulated Raman Scattering
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代谢研究

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Label-free DNA imaging in vivo with stimulated Raman scattering microscopy.
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免疫学

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植物学

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材料科学

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Operando and three-dimensional visualization of anion depletion and lithium growth by stimulated Raman scattering microscopy
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共振标签

Raman Imaging of Small Biomolecules
Y. Shen, F. Hu, W. Min
Annu Rev Biophys. 2019 May 6;48:347-369. doi: 10.1146/annurev-biophys-052118-115500. Epub 2019 Mar 20.

Spectral tracing of deuterium for imaging glucose metabolism
L. Zhang, L. Shi, Y. Shen, Y. Miao, M. Wei, N. Qian, Y. Liu, and W. Min
Nat Biomed Eng. 2019 May; 3(5): 402–413. Published online 2019 Apr 29. doi: 10.1038/s41551-019-0393-4

Optical imaging of metabolic dynamics in animals
L. Shi, C.Zheng, Y. Shen, Z. Chen, E.S. Silveira, L. Zhang, M. Wei, C. Liu, C. de Sena-Tomas, K. Targoff and W. Min
Nat Commun. 2018; 9: 2995. Published online 2018 Aug 6. doi: 10.1038/s41467-018-05401-3

Metabolic activity induces membrane phase separation in endoplasmic reticulum.
Shen Y, Zhao Z, Zhang L, Shi L, Shahriar S, Chan RB, Di Paolo G and Min W.
PNAS 2017, 114(51):13394-13399. DOI: 10.1073/pnas.1712555114

Supermultiplexed optical imaging and barcoding with engineered polyynes.
Hu F, Zeng C, Long R, Miao Y, Wei L, Xu Q, Min W.
Nat Methods. 2018 Mar;15(3):194-200. doi: 10.1038/nmeth.4578. Epub 2018 Jan 15.

Super-multiplex vibrational imaging
Lu Wei, Zhixing Chen, Lixue Shi, Rong Long, Andrew V. Anzalone, Luyuan Zhang, Fanghao Hu, Rafael Yuste, Virginia W. Cornish and Wei Min
Nature. 2017 Apr 27; 544(7651): 465–470. Published online 2017 Apr 19. doi:  10.1038/nature22051



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