{"id":20688,"date":"2021-05-31T16:35:08","date_gmt":"2021-05-31T10:50:08","guid":{"rendered":"https:\/\/www.revoscience.com\/en\/?p=20688"},"modified":"2021-05-31T16:35:11","modified_gmt":"2021-05-31T10:50:11","slug":"new-light-sheet-microscopy-unit-reduced-photodamage","status":"publish","type":"post","link":"https:\/\/www.revoscience.com\/en\/new-light-sheet-microscopy-unit-reduced-photodamage\/","title":{"rendered":"New light-sheet microscopy unit reduced photodamage"},"content":{"rendered":"\n
\"\"<\/figure>\n\n\n\n

A research group led by Takashi Saito, of the Ehime University Graduate School of Medicine, developed a 2-photon excitation light-sheet fluorescence microscope which lowers phototoxicity, extends the field of view, and heightens spatial resolution. <\/p>\n\n\n\n

This microscope, when used for the observation of medaka fish, made it possible to observe the whole body of the embryo at a cellular level resolution without affecting the growth of the fish over a three-day span of embryonic development. This result was published in the scientific journal “Nature Communications” (Springer Nature).<\/p>\n\n\n\n

According to researcher, Fluorescence microscope is widely used in the field of life science to observe molecules inside a cell in a non-invasive way. Light-sheet fluorescence microscopy makes it possible to record three-dimensional images with high acquisition speed and high spatial resolution. However, in conventional light-sheet microscopes, it is difficult to limit photodamage to living tissues, and also difficult to simultaneously achieve wide FOV and high (cell level) spatial resolution.<\/p>\n\n\n\n

In the process of development of a two-photon excitation wide-field light-sheet microscope, researchers were utilized the two-photon excitation phenomenon as a key to solving this problem. The two-photon excitation microscope with infrared lasers enables gentle (low phototoxic) imaging of living organisms. <\/p>\n\n\n\n

\"\"<\/figure>\n\n\n\n

However, because the light must be focused on a narrow range to induce two-photon excitation, the excitation range (in light-sheet microscope, the field of view) is narrow. In order to solve this, we developed simple illumination optics unit with a Bessel beam that expands the laser propagation range in the direction of the optic axis (Fig. 1A). <\/p>\n\n\n\n

This unit can stretch the beam length to 600-1000 \u03bcm while maintaining a 2-3 \u03bcm axial resolution when using a 10x magnification NA0.3 objective lens. Using this optical unit, we constructed a two-photon excitation light-sheet microscopy (Fig. 1B), which makes it possible to perform whole-body imaging of medaka larvae with cellular resolution (Fig. 1C).<\/p>\n\n\n\n

Application to live imaging<\/strong><\/p>\n\n\n\n

The medaka is widely used as a model organism for vertebrate. It is suitable for fluorescent imaging since it is small and transparent. To evaluate the applicability of our microscope for use on living organisms, they performed a phototoxicity assessment. <\/p>\n\n\n\n

This revealed reduced photodamage compared with the conventional Gaussian beam light-sheet sheet microscope. It is therefore suggested to be suitable for long-term live imaging. We then applied long-term time-lapse imaging of the transgenic medaka in which lymphatic endothelium is labelled with green fluorescent protein, and succeeded in live imaging over three days at intervals of 5 minutes (Fig. 2).<\/p>\n\n\n\n

In this study, they developed a new high performance light-sheet fluorescence microscope. Using this technology, they can observe almost all the embryonic growth processes of medaka fish with high cellular resolution over the whole body of the fish. <\/p>\n\n\n\n

This technology is expected to contribute to the molecular level understanding of embryonic development, the elucidation of pathogenesis for lifestyle-related diseases, and to further the technology of drug development.<\/p>\n","protected":false},"excerpt":{"rendered":"

A research group led by Takashi Saito, of the Ehime University Graduate School of Medicine, developed a 2-photon excitation light-sheet fluorescence microscope<\/p>\n","protected":false},"author":2,"featured_media":20689,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[17,28],"tags":[],"class_list":["post-20688","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-research","category-techbiz"],"featured_image_urls":{"full":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",1140,509,false],"thumbnail":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development-200x200.jpg",200,200,true],"medium":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development-675x301.jpg",675,301,true],"medium_large":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development-768x343.jpg",750,335,true],"large":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development-675x301.jpg",675,301,true],"1536x1536":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",1140,509,false],"2048x2048":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",1140,509,false],"ultp_layout_landscape_large":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",1140,509,false],"ultp_layout_landscape":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",870,388,false],"ultp_layout_portrait":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",600,268,false],"ultp_layout_square":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",600,268,false],"newspaper-x-single-post":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development-760x490.jpg",760,490,true],"newspaper-x-recent-post-big":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development-550x360.jpg",550,360,true],"newspaper-x-recent-post-list-image":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development-95x65.jpg",95,65,true],"web-stories-poster-portrait":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",640,286,false],"web-stories-publisher-logo":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",96,43,false],"web-stories-thumbnail":["https:\/\/www.revoscience.com\/en\/wp-content\/uploads\/2021\/05\/lymphatic-vessel-development.jpg",150,67,false]},"author_info":{"info":["RevoScience"]},"category_info":"Research<\/a> Tech<\/a>","tag_info":"Tech","comment_count":"0","_links":{"self":[{"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/posts\/20688","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/comments?post=20688"}],"version-history":[{"count":0,"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/posts\/20688\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/media\/20689"}],"wp:attachment":[{"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/media?parent=20688"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/categories?post=20688"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.revoscience.com\/en\/wp-json\/wp\/v2\/tags?post=20688"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}