Light needle microscopy with spatially transposed detection for axially resolved volumetric imaging

The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence microscopy has been widely employed for this purpose; however, a volumetric image is constructed by two-dimensional image stacking with a varying observation...

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Published inScientific reports Vol. 9; no. 1; pp. 11687 - 10
Main Authors Kozawa, Yuichi, Sato, Shunichi
Format Journal Article
LanguageEnglish
Published England Nature Publishing Group 12.08.2019
Nature Publishing Group UK
Subjects
Confocal microscopy
Fluorescence microscopy
Microscopy
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Abstract The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence microscopy has been widely employed for this purpose; however, a volumetric image is constructed by two-dimensional image stacking with a varying observation plane, ultimately limiting the acquisition speed. Here we propose a method enabling axially resolved volumetric imaging without a moving observation plane in the framework of laser scanning microscopy. A scanning light needle spot with an extended focal depth provides excitation, which normally produces a deep focus image with a loss of depth information. In our method, the depth information is retrieved from transposed lateral information on an array detector by utilising non-diffracting and self-bending characteristics imposed on fluorescent signals. This technique, implemented in two-photon microscopy, achieves truly volumetric images constructed from a single raster scan of a light needle, which has the capability to significantly reduce the acquisition time.
AbstractList The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence microscopy has been widely employed for this purpose; however, a volumetric image is constructed by two-dimensional image stacking with a varying observation plane, ultimately limiting the acquisition speed. Here we propose a method enabling axially resolved volumetric imaging without a moving observation plane in the framework of laser scanning microscopy. A scanning light needle spot with an extended focal depth provides excitation, which normally produces a deep focus image with a loss of depth information. In our method, the depth information is retrieved from transposed lateral information on an array detector by utilising non-diffracting and self-bending characteristics imposed on fluorescent signals. This technique, implemented in two-photon microscopy, achieves truly volumetric images constructed from a single raster scan of a light needle, which has the capability to significantly reduce the acquisition time.
Abstract The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence microscopy has been widely employed for this purpose; however, a volumetric image is constructed by two-dimensional image stacking with a varying observation plane, ultimately limiting the acquisition speed. Here we propose a method enabling axially resolved volumetric imaging without a moving observation plane in the framework of laser scanning microscopy. A scanning light needle spot with an extended focal depth provides excitation, which normally produces a deep focus image with a loss of depth information. In our method, the depth information is retrieved from transposed lateral information on an array detector by utilising non-diffracting and self-bending characteristics imposed on fluorescent signals. This technique, implemented in two-photon microscopy, achieves truly volumetric images constructed from a single raster scan of a light needle, which has the capability to significantly reduce the acquisition time.
ArticleNumber 11687
Author Kozawa, Yuichi
Sato, Shunichi
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  givenname: Yuichi
  orcidid: 0000-0002-9886-9083
  surname: Kozawa
  fullname: Kozawa, Yuichi
  email: y.kozawa@tohoku.ac.jp
  organization: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, 980-8577, Japan. y.kozawa@tohoku.ac.jp
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  givenname: Shunichi
  surname: Sato
  fullname: Sato, Shunichi
  organization: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, 980-8577, Japan
BackLink https://www.ncbi.nlm.nih.gov/pubmed/31406209$$D View this record in MEDLINE/PubMed
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  publication-title: Nat. Methods
  doi: 10.1038/nmeth.f.388
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Snippet The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence microscopy has...
Abstract The demand for rapid three-dimensional volumetric imaging is increasing in various fields, including life science. Laser scanning fluorescence...
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SubjectTerms Confocal microscopy
Fluorescence microscopy
Microscopy
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Title Light needle microscopy with spatially transposed detection for axially resolved volumetric imaging
URI https://www.ncbi.nlm.nih.gov/pubmed/31406209
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