2015年
2D simultaneous spatial and temporal focusing multiphoton microscopy for fast volume imaging with improved sectioning ability
MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES XV
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- 巻
- 9329
- 号
- 記述言語
- 英語
- 掲載種別
- 研究論文(国際会議プロシーディングス)
- DOI
- 10.1117/12.2078093
- 出版者・発行元
- SPIE-INT SOC OPTICAL ENGINEERING
Simultaneous spatial and temporal focusing (SSTF) multiphoton microscopy offers us widefield imaging with sectioning ability. As extending the idea to 2D SSTF, people can utilize a 2D spectral disperser. In this study, we use a 2D spectral disperser via a virtually-imaged phased-array (VIPA) and a diffraction grating to fulfill the back aperture of objective lens with a spectrum matrix. This offers us an axial resolution enhanced by a factor of similar to 1.7 compared with conventional SSTF microscopy. Furthermore, the small free spectral range (FSR) of VIPA will reduce the temporal self-imaging effect around out-of-focus region and thus will reduce the out-of-focus multiphoton excited fluorescence (MPEF) signal of 2D SSTF microscopy. We experimentally show that inside a sample with dense MPEF, the contrast of the sectioning image is increased in our 2D SSTF microscope compared with SSTF microscope. In our microscope, we use a 1 kHz chirped amplification laser, a piezo stage and a sCMOS camera integrated with 2D SSTF to realize high speed volume imaging at a speed of 50 volumes per second as well as improved sectioning ability. Volume imaging of Brownian motions of fluorescent beads as small as 1 mu m has been demonstrated. Not only the lateral motion but also the axial motion could be traced.
- リンク情報
- ID情報
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- DOI : 10.1117/12.2078093
- ISSN : 0277-786X
- Web of Science ID : WOS:000354164400038