2D simultaneous spatial and temporal focusing multiphoton microscopy for fast volume imaging with improved sectioning ability

Qiyuan Song, Keisuke Isobe, Kenichi Hirosawa, Katsumi Midorikawa, Fumihiko Kannari

    研究成果: Conference contribution

    2 被引用数 (Scopus)

    抄録

    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 ∼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 1μm has been demonstrated. Not only the lateral motion but also the axial motion could be traced.

    本文言語English
    ホスト出版物のタイトルMultiphoton Microscopy in the Biomedical Sciences XV
    編集者Karsten Konig, Peter T. C. So, Ammasi Periasamy
    出版社SPIE
    ISBN(電子版)9781628414196
    DOI
    出版ステータスPublished - 2015 1 1
    イベントMultiphoton Microscopy in the Biomedical Sciences XV - San Francisco, United States
    継続期間: 2015 2 82015 2 10

    出版物シリーズ

    名前Progress in Biomedical Optics and Imaging - Proceedings of SPIE
    9329
    ISSN(印刷版)1605-7422

    Other

    OtherMultiphoton Microscopy in the Biomedical Sciences XV
    国/地域United States
    CitySan Francisco
    Period15/2/815/2/10

    ASJC Scopus subject areas

    • 電子材料、光学材料、および磁性材料
    • 生体材料
    • 原子分子物理学および光学
    • 放射線学、核医学およびイメージング

    フィンガープリント

    「2D simultaneous spatial and temporal focusing multiphoton microscopy for fast volume imaging with improved sectioning ability」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

    引用スタイル