原位细胞3D切割成像平台-CellSurgeon

原位细胞3D切割成像平台-CellSurgeon

德国LLS ROWIAK公司推出的CellSurgeon是一款精准、非接触的3D纳米激光活细胞显微成像切割系统。它独具特色的多光子切割技术,能够从细胞内或组织内的任意点开始切割,实现真正意义上的定点操作。并且CellSurgeon还配有MPM成像模块,能够实现实时的荧光标记或无标记成像,精准定位所需操作的部位和实时观测细胞动态变化。通过CellSurgeon研究者能够进行实时精准的活细胞、组织操作和观测,帮助研究者更好的研究原位细胞的生理活性。


应用领域

■  染色体切割

■  亚细胞器的实时观测切割

■  原位组织的单细胞分离

■  薄组织的显微切割

■  基于激光的光转染技术

高精度可视化的3D细胞荧光显微切割系统,能够胜任从亚细胞尺度到细胞团的切割任务。能够进行线粒体消融、染色体切割、微丝微管切割等试验。能够从组织块或细胞团、培养细胞中原位分选单个细胞。#

CellSurgeon切割原理


CellSurgeon将近红外超短脉冲激光器耦合到显微镜中,并利用高数值孔径物镜聚焦超短激光脉冲,仅在最小的聚焦体积内产生高强度能量引起多光子吸收,然后以非常精确的方式在活细胞中实现亚细胞水平的细胞结构可视化操作。由于几乎没有热能或机械能传递,靠近激光束紧焦点的细胞结构依旧保持完好无损。


切割方式


双光子切割 VS 单光子切割

可从组织中的任意部位开始切割


CellSurgeon的切割方式






■  动脉激光切割和成像

30 fps超短激光脉冲对小鼠血管的损伤


体内激光诱导血栓的三维重建,采用FITC-葡聚糖染色双光子成像监测激光损伤后血栓的形成情况


■  肌动蛋白丝的切割

用飞秒激光切割肌动蛋白细丝


■  有丝分裂纺锤体的亚细胞解剖

GBP标记的有丝分裂纺锤体,光漂白(A)和切割消融(B)


■  细胞器消融

不同功率激光对核的消融,激光消融前(A)和后(B)


线粒体消融,激光消融前()和后()


■  从细胞到组织的动态观测与切割


CellSurgeon能够胜任各种类型的切割任务,无论是的染色质还是活体组织,它都能很好地胜任。

该设备可以兼容多种型号的显微镜,并且支持显微操作针等配件,能够在切割后实现对切割部分的转移。


从细胞团中切除的细胞并用微毛细管将提取细胞切出


固定的CHO的Alexa488标记的毒伞素切割


 

活U2OS细胞的FP635标记的肌动蛋白的切割


活GM-7373牛主动脉内皮细胞的

MitoTracker Orange ®的单线粒体消融实验

 

 

活GM-7373牛主动脉内皮细胞诱导凋亡实验




人发丝切割


 染色质切割


激光介导的细胞转染



白蚁的组织切割


小鼠活体血管切割


 ■  基于激光的原位细胞转染

 

无论是电转还是脂质体都需要先将细胞悬浮才能够进行入转染,但是Cellsurgeon能够在原位对细胞进行光穿孔实现细胞的转染,这种技术对于研究原位的细胞转染有着重大意义。


使用CellSurgeon对ZMTH3细胞进行转染pEGFP-C1、pEGFP-HMGA2、pEGFP-HMGB1经过48小时的图像


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31. Kermani O, Will F, Massow O, Oberheide U, Lubatschowski H. (2009) Echtzeitsteuerung einer Femtosekundenlaser Sub-Bowman-Keratomileusis an humanen Spenderaugen mittels optischer Kohärenztomographie. Klin Monatsbl Augenheilkd 226 965-969

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部分用户单位:

Bayer HealthCare, Cardiovascular Research

Leibniz University Hannover, Institute of Biophysics

Leibniz University Hannover, Institute for Quantum Optics-1,-2

University of Rostock, Division of Medicine Clinic III, Hematology, Oncology and Palliative Medicine

Institute for Bioprocessing and Analytical Measurement Techniques (iba)

mfd Diagnostics GmbH 







染色体切割

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