模塊化超分辨共聚焦顯微系統(tǒng)-LiveCodim

    傳統(tǒng)熒光顯微鏡受到光學(xué)衍射極限的影響，高的分辨率為200 nm，因此很難觀察細(xì)胞中的超微結(jié)構(gòu)。LiveCodim是一款模塊化超分辨共聚焦顯微系統(tǒng)，能夠適配絕大多數(shù)的倒置熒光顯微鏡，將現(xiàn)有的倒置顯微鏡升級(jí)成為具備寬場(chǎng)、共聚焦、超分辨三大模式的成像系統(tǒng)。LiveCodim通過(guò)獨(dú)特的錐形衍射顯微鏡—— 一種強(qiáng)大的波束成形器，能夠直接提供分辨率高達(dá)120 nm的實(shí)時(shí)活細(xì)胞超分辨共聚焦成像，同時(shí)無(wú)需對(duì)樣品進(jìn)行任何額外操作，結(jié)合其低光毒性，以及方便快捷的操作系統(tǒng)等優(yōu)勢(shì)，非常適合拍攝熒光成像。

產(chǎn)品優(yōu)勢(shì)

·  超高性價(jià)比：模塊化超分辨，節(jié)省成本，兼容絕大多數(shù)倒置顯微鏡

·  xy軸超高分辨率：<120 nm

·  z軸深度成像：具備z-stack成像能力，高成像深度50 μm

·  活細(xì)胞成像：低光毒性和光漂白性，適合活細(xì)胞成像

·  制樣簡(jiǎn)單：樣品無(wú)需特殊處理，無(wú)需特殊染料

·  全自動(dòng)軟件：全自動(dòng)調(diào)節(jié)各種參數(shù)，簡(jiǎn)單易上手

主要參數(shù)

·  xy軸分辨率：< 120 nm

·  z軸分辨率：< 500 nm

·  z軸成像深度：50 μm

·  成像視野：共聚焦模式下80 μm * 80 μm，超分辨模式下： 50 μm * 50 μm

·  成像模式：寬場(chǎng)、共聚焦、LiveCodim超分辨

·  四色成像通道：405 nm, 488 nm, 561 nm, 640 nm (根據(jù)需求可增加)

測(cè)試數(shù)據(jù)

1. MDCK細(xì)胞中線粒體的動(dòng)態(tài)變化

2.  Hela胞的微管寬場(chǎng)，共聚焦，LiveCodim超分辨成像

3.  細(xì)胞分裂中期的COS-7細(xì)胞3D多色超分辨成像

4.  植物細(xì)胞成像：觀測(cè)鈴蘭草的根莖

5.  天然免疫分子TRIM5α作用機(jī)制研究

    天然免疫分子TRIM5α蛋白是人類基因中決定疾病的易感性和發(fā)病速度的重要因素，其抗病毒活性通常通過(guò)小泛素相關(guān)修飾物(SUMO)調(diào)節(jié)，但是具體的作用機(jī)制仍有待進(jìn)一步研究。LiveCodim超分辨圖像揭示了TRIM5α主要分布在肌小管的核膜上，同時(shí)與存在于核孔的細(xì)胞質(zhì)絲上的RanGTPase激活蛋白R(shí)anGAP1有明顯的共定位現(xiàn)象，和主要定位于核籃上的蛋白Nup153無(wú)明顯共定位，說(shuō)明TRIM5α主要定位于這類細(xì)胞的胞質(zhì)面。

部分發(fā)表文章

[1] Fernandez, Juliette, et al. "Transportin-1 binds to the HIV-1 capsid via a nuclear localization signal and triggers uncoating." Nature microbiology 4.11 (2019): 1840-1850.

[2] Vargas, Jessica Y., et al. "The Wnt/Ca2+ pathway is involved in interneuronal communication mediated by tunneling nanotubes." The EMBO journal 38.23 (2019): e101230.

[3] Maarifi, Ghizlane, et al. "RanBP2 regulates the anti-retroviral activity of TRIM5α by SUMOylation at a predicted phosphorylated SUMOylation motif." Communications biology 1.1 (2018): 1-11.

[4] Garita-Hernandez, Marcela, et al. "Optogenetic light sensors in human retinal organoids." Frontiers in neuroscience 12 (2018): 789.

[5] Getz, Angela M., et al. "Tumor suppressor menin is required for subunit-specific nAChR α5 transcription and nAChR-dependent presynaptic facilitation in cultured mouse hippocampal neurons." Scientific reports 7.1 (2017): 1-16.

[6] Portilho, Débora M., Roger Persson, and Nathalie Arhel. "Role of non-motile microtubule-associated proteins in virus trafficking." Biomolecular concepts 7.5-6 (2016): 283-292.

[7] Pagliuso, Alessandro, et al. "A role for septin 2 in Drp1‐mediated mitochondrial fission." EMBO reports 17.6 (2016): 858-873.

[8] Fallet, Clement, and Gabriel Y. Sirat. "Achromatization of conical diffraction: application to the generation of a polychromatic optical vortex." Optics letters 41.4 (2016): 769-772.

[9] Fallet, Clement, et al. "Accurate axial localization by conical diffraction beam shaping generating a dark-helix PSF." Single Molecule Spectroscopy and Superresolution Imaging IX. Vol. 9714. International Society for Optics and Photonics, 2016.

[10] Fallet, Clement, Arvid Lindberg, and Gabriel Y. Sirat. "Generating 3D depletion distribution in an achromatic single-channel monolithic system." Single Molecule Spectroscopy and Superresolution Imaging IX. Vol. 9714. International Society for Optics and Photonics, 2016.

[11] Fallet, Clément, et al. "A new method to achieve tens of nm axial super-localization based on conical diffraction PSF shaping." Single Molecule Spectroscopy and Superresolution Imaging VIII. Vol. 9331. International Society for Optics and Photonics, 2015.

[12] Caron, Julien, et al. "Conical diffraction illumination opens the way for low phototoxicity super-resolution imaging." Cell adhesion & migration 8.5 (2014): 430-439.

[13] Fallet, Clément, et al. "Conical diffraction as a versatile building block to implement new imaging modalities for superresolution in fluorescence microscopy." Nanoimaging and Nanospectroscopy II. Vol. 9169. International Society for Optics and Photonics, 2014.

[14] Rosset, Sybille, Clement Fallet, and Gabriel Y. Sirat. "Focusing by a high numerical aperture lens of distributions generated by conical diffraction." Optics letters 39.23 (2014): 6569-6572.

用戶單位  

法國(guó)巴斯德研究所

蒙彼利埃大學(xué)