AbstractLightsheet microscopy has evolved to be a fast and practical tool for 3D imaging biological samples and is becoming a common instrument in core facilities. The Ultramicroscope II (UMII) from LaVision BioTec (A Miltenyi Biotec Company) has provided a user-friendly commercial solution for large scale imaging. It uses cylindrical lenses to generate the lightsheets enabling to image larger samples without the steric limitations that other lightsheet instrumentation pose. A requirement of this technique is to optically clear the samples a process that involves various chemicals, all of them can all be imaged in the same platform without further specialized lenses or containers. One of the greatest advantages of the UMII is the flexibility it provides in terms of lenses that can be mounted on the scope. In this work, we compare the use of the UMII 2x with zoom body to the use of the 12x lens in terms of performance in image quality, area coverage and speed. To give further options we have 3D printed custom sample holders in varied materials that allow for mounting larger specimens. To cover full specimens, we will use the mosaic function, explore different stitching strategies, and image quality enhancements. This work will provide guidelines for users interested in different scale imaging and speed alternative to other microscopy techniques and will help in decision making for lighsteet microscopy users. The workflow proposed here will help other lightsheet users and core facilities with their imaging pipelines.
University of Melbourne
Bio available soon