Inclined selective plane illumination microscopy adaptor for conventional microscopes
Article first published online: 27 JUN 2012
Copyright © 2012 Wiley Periodicals, Inc.
Microscopy Research and Technique
Volume 75, Issue 11, pages 1461–1466, November 2012
How to Cite
Cutrale, F. and Gratton, E. (2012), Inclined selective plane illumination microscopy adaptor for conventional microscopes. Microsc. Res. Tech., 75: 1461–1466. doi: 10.1002/jemt.22089
- Issue published online: 22 OCT 2012
- Article first published online: 27 JUN 2012
- Manuscript Accepted: 18 MAY 2012
- Manuscript Received: 4 APR 2012
- National Center for Research Resources. Grant Number: 5P41RR003155
- National Institute of General Medical Sciences, Divisions of the National Institutes of Health. Grant Numbers: 8P41GM103540, 5P50 GM076516
- SPIM microscopy;
- light sheet
Driven by the biological sciences, there is an increased need for imaging modalities capable of live cell imaging with high spatial and temporal resolution. To achieve this goal in a comprehensive manner, three-dimensional acquisitions are necessary. Ideal features of a modern microscope system should include high imaging speed, high contrast ratio, low photo-bleaching and photo-toxicity, good resolution in a 3D context, and mosaic acquisition for large samples. Given the importance of collecting data in live sample further increases the technical challenges required to solve these issues. This work presents a practical version of a microscopy method, Selective Plane Illumination Microscopy re-introduced by Huisken et al. (Science2004,305,1007–1009). This method is gaining importance in the biomedical field, but its use is limited by difficulties associated with unconventional microscope design which employs two objectives and a particular kind of sample preparation needed to insert the sample between the objectives. Based on the selective plane illumination principle but with a design similar to the Total Internal Reflection Fluorescence microscope, Dunsby (Dunsby, Opt Express2008,16,20306–20316) demonstrated the oblique plane microscope (OPM) using a single objective which uses conventional sample preparation protocols. However, the Dunsby instrument was not intended to be part of a commercial microscope. In this work, we describe a system with the advantages of OPM and that can be used as an adaptor to commonly used microscopes, such as IX-71 Olympus, simplifying the construction of the OPM and increasing performance of a conventional microscope. We named our design inclined selective plane illumination microscope (iSPIM). Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.