A digital light-sheet microscope for the analysis of biological processes under almost natural conditions

多年来，生物成像一直在朝着采用越来越多的生理相关系统的实验方向发展，新技术正在使这种实验方法逐渐可行。这样的系统，包括整个模型生物体，组织外植体，和三维（3D）细胞培养物，必须以最小化扰动的方式成像，以保持其作为实验模型的生理完整性。光诱导的光损伤和光毒性长期以来一直是生物成像领域的挑战，并且可以对所有生物组织水平的健康和功能产生相当大的影响。因此，对如此大而敏感的样本进行成像需要一种有效的3D成像方法，以减少样本暴露在光线下的时间。光片显微镜代表了一种平面照明技术，该技术彻底改变了生物样本的光学成像方式。基本上，光片技术通过将照明和检测光学路径解耦而成为可能，从而允许优化曝光和信号收集效率的新颖照明策略。利用光片显微镜，可以将较低的照明强度与平面照明结合使用，以最小的样品曝光提供改善的信号，从而能够以高帧率和长时间成像。已经从这种成像模式中受益匪浅的领域包括发育生物学、神经生物学、药物发现、植物生物学等。然而，定制的光片显微镜可能非常难以使用，通常需要两个或更多物镜、非标准的样品制备协议、困难的对准程序以及精心规划的工作流程来处理这些技术产生的大量数据。我们的目标是购买一个商业光片显微镜，这是最简单，最直观，最可靠的系统，结合一个简单的数据处理和分析平台，可以使研究人员在几乎自然的条件下研究生物过程。

Biological imaging has been moving consistently towards experimentation employing increasingly physiologically relevant systems for many years, and new technologies are making such experimental approaches progressively feasible. Such systems, comprising whole model organisms, tissue explants, and three-dimensional (3D) cell cultures, must be imaged in a way that minimises perturbations in order to maintain their physiological integrity as experimental models. Light-induced photodamage and phototoxicity have long been challenging in the field of biological imaging and can have quite a dramatic effect on health and function at all biological levels of organisation. Imaging such large and sensitive specimens thus demands an efficient approach to 3D imaging that lessens exposure of the sample to light.The light-sheet microscope stands for a planar illumination technique that has revolutionised how optical imaging of biological specimens can be performed. Basically, the light-sheet technique is made possible by decoupling the illumination and detection optical pathways, allowing for novel illumination strategies that optimise the light exposure and signal collection efficiency. With light-sheet microscopy, lower illumination intensities can be used in combination with planar illumination to provide improved signal with minimal sample light exposure, enabling imaging with high frame rates and over long periods of time. Fields that have already benefited considerably from such an imaging modality include developmental biology, neurobiology, drug discovery, plant biology, and beyond.However, custom-made light-sheet microscopes can be very difficult to use, usually requiring two or more objectives, non-standard sample preparation protocols, difficult alignment procedures, and a well-planned workflow for dealing with the massive amounts of data generated by such techniques. Our aim is to purchase a commercial light-sheet microscope that is the simplest, most intuitive and most reliable system available that in combination with a straightforward data handling and analysis platform can empower researchers to investigate biological processes under almost natural conditions.