Tosi Sébastien, Lladó Anna, Bardia Lídia, Rebollo Elena, Godo Anna, Stockinger Petra, Colombelli Julien
Institute for Research in Biomedicine, IRB Barcelona, Barcelona Institute of Science and Technology, BIST, Barcelona, Spain.
Molecular Imaging Platform, Molecular Biology institute of Barcelona IBMB-CSIC, Barcelona, Spain.
Front Bioinform. 2021 Mar 18;1:627626. doi: 10.3389/fbinf.2021.627626. eCollection 2021.
We developed AutoscanJ, a suite of ImageJ scripts enabling to image targets of interest by automatically driving a motorized microscope at the corresponding locations. For live samples, our software can sequentially detect biological events from their onset and further image them at high resolution, an action that would be impractical by user operation. For fixed samples, the software can dramatically reduce the amount of data acquired and the acquisition duration in situations where statistically few targets of interest are observed per field of view. AutoScanJ is compatible with motorized fluorescence microscopes controlled by Leica LAS AF/X or Micro-Manager. The software is straightforward to set up and new custom image analysis workflows to detect targets of interest can be simply implemented and shared with minimal efforts as independent ImageJ macro functions. We illustrate five different application scenarios with the system ranging from samples fixed on micropatterned surfaces to live cells undergoing several rounds of division. The target detection functions for these applications are provided and can be used as a starting point and a source of inspiration for new applications. Overall, AutoScanJ helps to optimize microscope usage by autonomous operation, and it opens up new experimental avenues by enabling the real-time detection and selective imaging of transient events in live microscopy.
我们开发了AutoscanJ,这是一套ImageJ脚本,能够通过在相应位置自动驱动电动显微镜来对感兴趣的目标进行成像。对于活样本,我们的软件可以从生物事件开始时就对其进行连续检测,并以高分辨率对其进一步成像,而这一操作若由用户手动进行则不切实际。对于固定样本,在每个视野中统计观察到的感兴趣目标较少的情况下,该软件可以显著减少采集的数据量和采集持续时间。AutoScanJ与由徕卡LAS AF/X或Micro-Manager控制的电动荧光显微镜兼容。该软件易于设置,并且可以轻松地将用于检测感兴趣目标的新的自定义图像分析工作流程作为独立的ImageJ宏函数来实现和共享。我们用该系统展示了五种不同的应用场景,从固定在微图案表面的样本到经历多轮分裂的活细胞。文中提供了这些应用的目标检测功能,可作为新应用的起点和灵感来源。总体而言,AutoScanJ通过自主操作有助于优化显微镜的使用,并通过在活细胞显微镜中实现对瞬态事件的实时检测和选择性成像开辟了新的实验途径。