双光子3D组织切割成像系统

双光子3D组织切割成像系统

                         ——OCT图像引导的组织和材料的非接触精确切割，更适合切小鼠胫骨等小鼠骨骼系统，牙齿的激光切片设备

应用领域

■  骨科方面，尤其是非脱钙硬组织和种植体界面研究

■  心脏病学和心血管研究与医学，尤其是软组织与生物材料和支架，钙化斑块研究

■  再生医学与组织工程学，尤其是植入物、支架等研究

■  口腔、面部和牙科医学，尤其是非脱钙硬组织，金属、陶瓷或聚合物植入物研究

■  耳鼻喉相关研究，如耳蜗、耳蜗植入物等

■  从小鼠到大型动物模型的临床前研究等

为何选用TissueSurgeon？

■  样本损失小：几乎连续切片非脱钙硬组织，无需大深度磨片，材料损失小；

■  难切割样本：硬组织、软组织、软硬结合组织切割，甚至脆弱的样本(如耳蜗)切割；

■  适合界面研究：种植体组织界面组织学（如牙钉、心血管支架、支架）；

■  无接触切割：无接触激光切割组织可避免挤压、划伤或裂纹等；

■  用3D切片方法可以沿着牙钉种植体-组织界面对特定部位的样本进行定向、温和的分离；

■  切割过程不会污染、灼伤或机械力损伤样品，可用于生物化学分析的无污染和无接触样品的制备；

■  用于组织工程的生物材料切割（如支架、聚四氟乙烯、水凝胶）；

■  组织，基质和材料的3D微结构切割；

■  薄切片厚度：硬组织切片10 µm；

■  切片速度：≥1 mm²/s；

■  光源类型：红外飞秒脉冲激光；

■  光学相干断层扫描（OCT）引导切割，可以测量样品尺寸和层厚，并能够高效定位到病变部位，直接对病变部位进行切割，大大提高了切割效率。

设备参数

TissueSurgeon产品升级！激光组织学的新维度：超大尺寸和可调节

可以实现最大6.6 cm样本切割

应用案例

■  TissueSurgeon可视化切片系统，实现边看边切

对于病理等多种研究来书，涉及到组织切片的内容， 困难的部分莫过于寻找病变部位。 相比一个完整组织来说， 有时候研究者所关注的部分仅仅是其中变异的一小部分组织的形态而已。 但是对于传统切片手段来说， 缺乏一种有效的手段来定位这个区域， 因此往往需要投入大量人力和物力去多次制样，大量切片来寻找这个部位。 TissueSurgeon 自身集成了适合深层组织细胞成像的光学相干断层扫描（OCT）成像功能， 帮助您直接定位到 ROI 区域。 让切片变得可视化， 实现更加和可控的切片。为研究者更加迅速直观的找到病变位置，大大提高了研究效率。

对含有金属钉的骨骼进行成OCT成像，并引导切片

■  原位细胞3D切割成像技术基于青鳉胚胎组织的单细胞提取

单细胞的原位组织提取一直以来都是一项十分困难的工作，这主要受制于组织之间连接致密难以消化，而机械力往往很难地将单个细胞与组织完整的分离。激光切割具有传统切割技术所难以匹及的切割精度，是目前一种比较理想的切割手段，因此围绕激光切割技术的相关显微产品也孕育而生，并在科研领域中越来越受到关注。但是激光切割也有其局限性，首先显微激光切割往往要从表面开始，无法对深层组织进行切割；另一方面激光的光源往往采用紫外激光光源，这种类型的光源很容易造成组织灼伤，从而影响切割下来样品的品质，因此激光切割的应用发展也受到了诸多限制。

如今ROWIAK公司推出的一款全新的单细胞分离系统有望解决这一难题。它采用了近红外双光子激光切割技术，在保留了激光切割精度优势的同时，采用近红外波长的激光从而避免了激光切中对组织灼烧的问题。因此能够实现的原位组织中的单个细胞的分离。

       青鳉是一种成熟的模式生物，常用于分析发育和发育过程中的细胞信号神经生物学研究。其中使用表达荧光蛋白的转基因胚胎是一种揭示胚胎发育的良好方法。随着基因技术的发展，研究者们越来越多地开始关注这些标记细胞中转录组中的信息。虽然单细胞测序技术发展迅速，但是从组织中获得单细胞的手段却十分有限。目前几乎没有手段能够直接在组织的原位上快速获取一个细胞，但是基于ROWIAK双光子切割技术，研究者成功地在这方面取得了一些进展。

青鳉胚胎中感知神经中表达mcherry的细胞成像

研究者为了研究青鳉感觉神经分泌细胞细胞群中特定表达m-cherry的转基因细胞的内部遗传信息，将ROWIAK双光子3D组织切割成像系统与传统的显微操作系统进行结合，成功实现了对目标细胞的原位分离。

研究者首先利用双光子3D组织切割成像系统对青鳉胚胎中的mcherry细胞进行了定位，然后根据其细胞群的形态设定了切割部位，随后系统根据预先设定的范围进行切割。待切割完成后使用玻璃微管移液器将目标的细胞部位直接取出，即获得了目标组织区域。这种方法能够在不破坏样品原位信息的情况下将感兴趣的部位直接的分离，这对于揭示生物体的基因表达情况具有着深远的意义。

从青鳉胚胎中分离特定表达mcherry的细胞团

参考文献：
Wittbrodt, J. et al. Medaka — a model organism from the Far East. Nature Reviews Genetics 3, 53-64.
Yamamoto, T. (ed.) MEDAKA (Killifish): Biology and strains. Yamamoto, T. (ed.) Keigaku Pub. Co., Tokyo, 1975, pp.365.
Kristin Tessmar-Raible et al.Removal of fluorescently-labeled sensory-neurosecretory cells from forebrain of transgenic Medaka embryos, focusonmicroscop. 2011.

测试数据

发表文章

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