Serir Rayane Hamdane, Deliot Aurelie, Kizilyaprak Caroline, Daraspe Jean, Walczak Christine, Canini Françoise, Leleu Amandine, Marco Sergio, Ronzon Frederic, Messaoudi Cedric
Multimodal Imaging Center, CNRS UAR2016, INSERM US43, Institut Curie, PSL Research University, Université Paris-Saclay, 91401 Orsay Cedex, France.
Sanofi, 69280 Marcy-l'Étoile, France.
Bioinform Adv. 2023 Sep 6;3(1):vbad119. doi: 10.1093/bioadv/vbad119. eCollection 2023.
FIB-SEM (Focused Ion Beam-Scanning Electron Microscopy) is a technique to generate 3D images of samples up to several microns in depth. The principle is based on the alternate use of SEM to image the surface of the sample (a few nanometers thickness) and of FIB to mill the surface of the sample a few nanometers at the time. In this way, huge stacks of images can thus be acquired.Although this technique has proven useful in imaging biological systems, the presence of some visual artifacts (stripes due to sample milling, detector saturation, charge effects, focus or sample drift, etc.) still raises some challenges for image interpretation and analyses.
With the aim of meeting these challenges, we developed a freeware (SEM3De) that either corrects artifacts with state-of-the-art approaches or, when artifacts are impossible to correct, enables the replacement of artifactual slices by an in-painted image created from adjacent non-artifactual slices. Thus, SEM3De improves the overall usability of FIB-SEM acquisitions.
SEM3De can be downloaded from https://sourceforge.net/projects/sem3de/ as a plugin for ImageJ.
聚焦离子束扫描电子显微镜(FIB-SEM)是一种用于生成深度达几微米的样品三维图像的技术。其原理基于交替使用扫描电子显微镜(SEM)对样品表面(厚度为几纳米)进行成像,以及聚焦离子束(FIB)每次对样品表面进行几纳米的铣削。通过这种方式,可以获取大量的图像堆栈。尽管该技术已被证明在生物系统成像中很有用,但一些视觉伪像(如样品铣削产生的条纹、探测器饱和、电荷效应、聚焦或样品漂移等)的存在仍然给图像解释和分析带来了一些挑战。
为了应对这些挑战,我们开发了一款免费软件(SEM3De),它既可以用最先进的方法校正伪像,也可以在无法校正伪像时,用由相邻无伪像切片创建的内插图像替换有伪像的切片。因此,SEM3De提高了FIB-SEM采集数据的整体可用性。
SEM3De可从https://sourceforge.net/projects/sem3de/ 作为ImageJ的插件下载。