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

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Citations (Scopus)

Abstract

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.

Original languageEnglish
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
PublisherSPIE
Volume9329
ISBN (Print)9781628414196
DOIs
Publication statusPublished - 2015
EventMultiphoton Microscopy in the Biomedical Sciences XV - San Francisco, United States
Duration: 2015 Feb 82015 Feb 10

Other

OtherMultiphoton Microscopy in the Biomedical Sciences XV
CountryUnited States
CitySan Francisco
Period15/2/815/2/10

Fingerprint

Microscopy
Microscopic examination
microscopy
Imaging techniques
microscopes
phased arrays
Microscopes
Fluorescence
fluorescence
gratings (spectra)
beads
Brownian movement
Diffraction gratings
Lenses
apertures
cameras
lenses
high speed
Lasers
Amplification

Keywords

  • nonlinear microscopy
  • three-dimensional microscopy
  • wide-field microscopy

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

Cite this

Song, Q., Isobe, K., Hirosawa, K., Midorikawa, K., & Kannari, F. (2015). 2D simultaneous spatial and temporal focusing multiphoton microscopy for fast volume imaging with improved sectioning ability. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 9329). [93292N] SPIE. https://doi.org/10.1117/12.2078093

2D simultaneous spatial and temporal focusing multiphoton microscopy for fast volume imaging with improved sectioning ability. / Song, Qiyuan; Isobe, Keisuke; Hirosawa, Kenichi; Midorikawa, Katsumi; Kannari, Fumihiko.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 9329 SPIE, 2015. 93292N.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Song, Q, Isobe, K, Hirosawa, K, Midorikawa, K & Kannari, F 2015, 2D simultaneous spatial and temporal focusing multiphoton microscopy for fast volume imaging with improved sectioning ability. in Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 9329, 93292N, SPIE, Multiphoton Microscopy in the Biomedical Sciences XV, San Francisco, United States, 15/2/8. https://doi.org/10.1117/12.2078093
Song Q, Isobe K, Hirosawa K, Midorikawa K, Kannari F. 2D simultaneous spatial and temporal focusing multiphoton microscopy for fast volume imaging with improved sectioning ability. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 9329. SPIE. 2015. 93292N https://doi.org/10.1117/12.2078093
Song, Qiyuan ; Isobe, Keisuke ; Hirosawa, Kenichi ; Midorikawa, Katsumi ; Kannari, Fumihiko. / 2D simultaneous spatial and temporal focusing multiphoton microscopy for fast volume imaging with improved sectioning ability. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 9329 SPIE, 2015.
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