{"id":198,"date":"2019-07-05T15:40:09","date_gmt":"2019-07-05T19:40:09","guid":{"rendered":"https:\/\/sites.bu.edu\/biomicroscopy\/?page_id=198"},"modified":"2019-07-15T09:41:41","modified_gmt":"2019-07-15T13:41:41","slug":"edof","status":"publish","type":"page","link":"https:\/\/sites.bu.edu\/biomicroscopy\/research\/edof\/","title":{"rendered":"Extended-depth-of-field microscopy"},"content":{"rendered":"<figure id=\"attachment287\" aria-describedby=\"caption-attachment287\" style=\"width: 450px\" class=\"wp-caption alignright\"><img loading=\"lazy\" src=\"\/biomicroscopy\/files\/2019\/07\/PSF-vs-EPSF.gif\" alt=\"\" class=\"wp-image-287 size-full\" width=\"440\" height=\"217\" \/><figcaption id=\"caption-attachment287\" class=\"wp-caption-text\">Standard vs. Extended PSF.<\/figcaption><\/figure>\n<p>Description: We present a wide-field fluorescence microscopy add-on that provides a fast, light-efficient extended depth-of-field (EDOF) using a deformable mirror with an update rate of 20 kHz. Out-of-focus contributions in the raw EDOF images are suppressed with a deconvolution algorithm derived directly from the microscope 3D optical transfer function. Demonstrations of the benefits of EDOF microscopy are shown with GCaMP-labeled mouse brain tissue.<\/p>\n<p>Significantly increased contrast can be obtained by combining EDOF imaging with targeted illumination. The latter strategy involves delivering illumination only to in-focus structure during the focal sweep, using illumination masks controlled by a fast DMD.<\/p>\n<figure id=\"attachment56\" aria-describedby=\"caption-attachment56\" style=\"width: 646px\" class=\"wp-caption aligncenter\"><img loading=\"lazy\" src=\"\/biomicroscopy\/files\/2019\/06\/Targeted-illumination-EDOF-636x305.png\" alt=\"\" class=\"wp-image-56 size-medium\" width=\"636\" height=\"305\" srcset=\"https:\/\/sites.bu.edu\/biomicroscopy\/files\/2019\/06\/Targeted-illumination-EDOF-636x305.png 636w, https:\/\/sites.bu.edu\/biomicroscopy\/files\/2019\/06\/Targeted-illumination-EDOF-768x368.png 768w, https:\/\/sites.bu.edu\/biomicroscopy\/files\/2019\/06\/Targeted-illumination-EDOF.png 967w\" sizes=\"(max-width: 636px) 100vw, 636px\" \/><figcaption id=\"caption-attachment56\" class=\"wp-caption-text\">Video-rate EDOF imaging. Conventional EDOF (using a tunable lens) is contrast enhanced by targeted illumination (using a DMD).<\/figcaption><\/figure>\n<ul>\n<li>S. Xiao, H. Tseng, H. Gritton, X. Han, J. Mertz, &#8220;Video-rate volumetric neuronal imaging using 3D targeted illumination&#8221;, Sci. Reports 8, 7921 (2018). <a href=\"https:\/\/www.nature.com\/articles\/s41598-018-26240-8\">link<\/a><\/li>\n<li>W. J. Shain, N. A. Vickers, J. Li, X. Han, T. Bifano, J. Mertz, \u201cAxial localization with modulated-illumination extended-depth-of-field microscopy\u201d, Biomed. Opt. Express 9, 1771-1782 (2018). <a href=\"https:\/\/www.osapublishing.org\/boe\/abstract.cfm?uri=boe-9-4-1771\">link<\/a><\/li>\n<li>W. J. Shain, N. A. Vickers, A. Negash, T. Bifano, A. Sentenac, J. Mertz, \u201cDual fluorescence-absorption deconvolution applied to extended-depth-of-field microscopy\u201d, Opt. Lett. 42, 4183-4186 (2017). <a href=\"https:\/\/www.osapublishing.org\/ol\/abstract.cfm?uri=ol-42-20-4183\">link<\/a><\/li>\n<li>W. J. Shain, N. A. Vickers, B. G. Goldberg, T. Bifano, J. Mertz, \u201cExtended depth of field microscopy with a high-speed deformable mirror\u201d, Opt. Lett. 42, 995-998 (2017). <a href=\"https:\/\/www.osapublishing.org\/ol\/abstract.cfm?uri=ol-42-5-995\">link<\/a><\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Description: We present a wide-field fluorescence microscopy add-on that provides a fast, light-efficient extended depth-of-field (EDOF) using a deformable mirror with an update rate of 20 kHz. Out-of-focus contributions in the raw EDOF images are suppressed with a deconvolution algorithm derived directly from the microscope 3D optical transfer function. Demonstrations of the benefits of EDOF [&hellip;]<\/p>\n","protected":false},"author":16427,"featured_media":0,"parent":98,"menu_order":17,"comment_status":"closed","ping_status":"closed","template":"page-templates\/profiles.php","meta":[],"_links":{"self":[{"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/pages\/198"}],"collection":[{"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/users\/16427"}],"replies":[{"embeddable":true,"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/comments?post=198"}],"version-history":[{"count":7,"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/pages\/198\/revisions"}],"predecessor-version":[{"id":447,"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/pages\/198\/revisions\/447"}],"up":[{"embeddable":true,"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/pages\/98"}],"wp:attachment":[{"href":"https:\/\/sites.bu.edu\/biomicroscopy\/wp-json\/wp\/v2\/media?parent=198"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}