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{"title"=>"Fluorescence cell imaging and manipulation using conventional halogen lamp microscopy", "type"=>"journal", "authors"=>[{"first_name"=>"Kazuo", "last_name"=>"Yamagata", "scopus_author_id"=>"35265370500"}, {"first_name"=>"Daisaku", "last_name"=>"Iwamoto", "scopus_author_id"=>"34569919100"}, {"first_name"=>"Yukari", "last_name"=>"Terashita", "scopus_author_id"=>"35197341900"}, {"first_name"=>"Chong", "last_name"=>"Li", "scopus_author_id"=>"25928640300"}, {"first_name"=>"Sayaka", "last_name"=>"Wakayama", "scopus_author_id"=>"7005358889"}, {"first_name"=>"Yoko", "last_name"=>"Hayashi-Takanaka", "scopus_author_id"=>"24335355600"}, {"first_name"=>"Hiroshi", "last_name"=>"Kimura", "scopus_author_id"=>"56735099000"}, {"first_name"=>"Kazuhiro", "last_name"=>"Saeki", "scopus_author_id"=>"7202259899"}, {"first_name"=>"Teruhiko", "last_name"=>"Wakayama", "scopus_author_id"=>"7006125572"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84863069586", "pui"=>"364218388", "scopus"=>"2-s2.0-84863069586", "issn"=>"19326203", "pmid"=>"22347500", "doi"=>"10.1371/journal.pone.0031638"}, "id"=>"f552c12e-da38-3c13-89bf-f155fe155761", "abstract"=>"Technologies for vitally labeling cells with fluorescent dyes have advanced remarkably. However, to excite fluorescent dyes currently requires powerful illumination, which can cause phototoxic damage to the cells and increases the cost of microscopy. We have developed a filter system to excite fluorescent dyes using a conventional transmission microscope equipped with a halogen lamp. This method allows us to observe previously invisible cell organelles, such as the metaphase spindle of oocytes, without causing phototoxicity. Cells remain healthy even after intensive manipulation under fluorescence observation, such as during bovine, porcine and mouse somatic cell cloning using nuclear transfer. This method does not require expensive epifluorescence equipment and so could help to reduce the science gap between developed and developing countries.", "link"=>"http://www.mendeley.com/research/fluorescence-cell-imaging-manipulation-using-conventional-halogen-lamp-microscopy", "reader_count"=>47, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>4, "Student > Doctoral Student"=>1, "Researcher"=>26, "Student > Ph. D. Student"=>6, "Other"=>2, "Student > Master"=>4, "Student > Bachelor"=>2, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>4, "Student > Doctoral Student"=>1, "Researcher"=>26, "Student > Ph. D. Student"=>6, "Other"=>2, "Student > Master"=>4, "Student > Bachelor"=>2, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>37, "Medicine and Dentistry"=>3, "Neuroscience"=>1, "Physics and Astronomy"=>1, "Chemistry"=>1, "Earth and Planetary Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Earth and Planetary Sciences"=>{"Earth and Planetary Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>37}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "Czech Republic"=>1, "Norway"=>1, "United States"=>3, "Japan"=>6, "United Kingdom"=>2}, "group_count"=>0}

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• http://doi.org/10.1089/cell.2013.0051
• http://doi.org/10.1089/cell.2014.0086
• http://doi.org/10.1039/C5AN01423A
• http://doi.org/10.1038/s41540-020-00152-8
• http://doi.org/10.1089/cell.2016.0026
• http://doi.org/10.1002/dvg.23277

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• {"files"=>["https://ndownloader.figshare.com/files/685500"], "description"=>"<p>The antibody–phycoerythrin conjugate diffused immediately after microinjection into mouse oocytes (A) but the conjugate could bind to the MII chromosome array within a few minutes (B, C). Five minutes later, the chromosomes became sufficiently bright for enucleation (D). The brightness of MII chromosomes was measured by comparison with the cytoplasm; the luminance increased with time up to 8 min (E). Imaging could reveal individual chromosomes (F, G). Different concentrations of the phycoerythrin conjugate were microinjected into oocytes (H–K). The staining intensity was proportional to the concentration of conjugate used and 75 µg/mL of antibody was the minimum needed for clear observation. Fading of the phycoerythrin label was examined (L–O). (L, M) Images of phycoerythrin-injected oocytes observed using a conventional fluorescence microscope faded within 30 s. (M, N) When these samples were observed using the halogen light system, the image did not fade even when observed continuously for over 10 min.</p>", "links"=>[], "tags"=>["mii", "spindle", "phycoerythrin"], "article_id"=>355981, "categories"=>["Physics", "Biotechnology", "Science Policy", "Genetics", "Developmental Biology"], "users"=>["Kazuo Yamagata", "Daisaku Iwamoto", "Yukari Terashita", "Chong Li", "Sayaka Wakayama", "Yoko Hayashi-Takanaka", "Hiroshi Kimura", "Kazuhiro Saeki", "Teruhiko Wakayama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031638.g003", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Staining_of_MII_spindle_using_phycoerythrin_and_observation_/355981", "title"=>"Staining of MII spindle using phycoerythrin and observation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-08 01:39:41"}
• {"files"=>["https://ndownloader.figshare.com/files/685775"], "description"=>"<p>Production of cloned mice using phycoerythrin and antibody injected oocyte.</p>", "links"=>[], "tags"=>["cloned", "mice", "phycoerythrin", "antibody", "injected"], "article_id"=>356251, "categories"=>["Physics", "Biotechnology", "Science Policy", "Genetics", "Developmental Biology"], "users"=>["Kazuo Yamagata", "Daisaku Iwamoto", "Yukari Terashita", "Chong Li", "Sayaka Wakayama", "Yoko Hayashi-Takanaka", "Hiroshi Kimura", "Kazuhiro Saeki", "Teruhiko Wakayama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031638.t002", "stats"=>{"downloads"=>4, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Production_of_cloned_mice_using_phycoerythrin_and_antibody_injected_oocyte_/356251", "title"=>"Production of cloned mice using phycoerythrin and antibody injected oocyte.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-02-08 01:44:11"}
• {"files"=>["https://ndownloader.figshare.com/files/349084", "https://ndownloader.figshare.com/files/349127", "https://ndownloader.figshare.com/files/349166", "https://ndownloader.figshare.com/files/349193", "https://ndownloader.figshare.com/files/349242", "https://ndownloader.figshare.com/files/349294", "https://ndownloader.figshare.com/files/349387"], "description"=>"<div><p>Technologies for vitally labeling cells with fluorescent dyes have advanced remarkably. However, to excite fluorescent dyes currently requires powerful illumination, which can cause phototoxic damage to the cells and increases the cost of microscopy. We have developed a filter system to excite fluorescent dyes using a conventional transmission microscope equipped with a halogen lamp. This method allows us to observe previously invisible cell organelles, such as the metaphase spindle of oocytes, without causing phototoxicity. Cells remain healthy even after intensive manipulation under fluorescence observation, such as during bovine, porcine and mouse somatic cell cloning using nuclear transfer. This method does not require expensive epifluorescence equipment and so could help to reduce the science gap between developed and developing countries.</p> </div>", "links"=>[], "tags"=>["fluorescence", "imaging", "manipulation", "halogen", "microscopy"], "article_id"=>129033, "categories"=>["Physics", "Biotechnology", "Science Policy", "Genetics", "Developmental Biology"], "users"=>["Kazuo Yamagata", "Daisaku Iwamoto", "Yukari Terashita", "Chong Li", "Sayaka Wakayama", "Yoko Hayashi-Takanaka", "Hiroshi Kimura", "Kazuhiro Saeki", "Teruhiko Wakayama"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0031638.s001", "https://dx.doi.org/10.1371/journal.pone.0031638.s002", "https://dx.doi.org/10.1371/journal.pone.0031638.s003", "https://dx.doi.org/10.1371/journal.pone.0031638.s004", "https://dx.doi.org/10.1371/journal.pone.0031638.s005", "https://dx.doi.org/10.1371/journal.pone.0031638.s006", "https://dx.doi.org/10.1371/journal.pone.0031638.s007"], "stats"=>{"downloads"=>15, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Fluorescence_Cell_Imaging_and_Manipulation_Using_Conventional_Halogen_Lamp_Microscopy/129033", "title"=>"Fluorescence Cell Imaging and Manipulation Using Conventional Halogen Lamp Microscopy", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-02-08 02:30:33"}
• {"files"=>["https://ndownloader.figshare.com/files/685344"], "description"=>"<p>The adapter is placed on the top of the condenser (A). This adapter has a diaphragm (B) and a gap between the filter and frame (C). (D) Bright field and (E) fluorescence of green fluorescent protein (GFP)-labeled embryonic stem (ES) cells in the ICM of a chimeric mouse blastocyst. When the diaphragm is opened, halogen light can pass around the filter and a merged image can be obtained (F). This filter adapter could also be useful in purifying rare cell populations. Typically, fluorescence-activated cell sorting is used for this purpose, but cannot be applied if samples do not contain sufficient cell numbers. We collected spermatogonia from neonatal mouse testes using a CD9 antibody labeled with phycoerythrin. The CD9-positive spermatogonia could be discriminated from other cells and we were able to collect them using a micromanipulator without the need for a mercury vapor lamp. (G) Spermatogonia were detected using a CD9 antibody–phycoerythrin conjugate and positively stained cells (arrows) were drawn into a micropipette using a micromanipulator (H).</p>", "links"=>[], "tags"=>["designed", "adapter"], "article_id"=>355829, "categories"=>["Physics", "Biotechnology", "Science Policy", "Genetics", "Developmental Biology"], "users"=>["Kazuo Yamagata", "Daisaku Iwamoto", "Yukari Terashita", "Chong Li", "Sayaka Wakayama", "Yoko Hayashi-Takanaka", "Hiroshi Kimura", "Kazuhiro Saeki", "Teruhiko Wakayama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031638.g002", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_newly_designed_filter_adapter_and_its_applications_/355829", "title"=>"The newly designed filter adapter and its applications.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-08 01:37:09"}
• {"files"=>["https://ndownloader.figshare.com/files/685625"], "description"=>"<p>The MII chromosomes of bovine oocytes are normally invisible by conventional microscopy because of dark lipid droplets in the ooplasm (A). After labeling with an H3S10ph antibody–phycoerythrin conjugate, the MII chromosomes could be recognized clearly and this allowed us to remove them along with minimal cytoplasm (B). MII chromosomes in porcine oocytes were detected by the same method (C). Some chromosomes (arrow) were left occasionally inside the cytoplasm of mouse oocytes after enucleation (D). Live and healthy cloned mice (brown) were obtained after enucleation using this method without any significant decrease in the success rate (E). Enucleation of the MII spindle from mouse oocytes with this system. (F) Before, (G) during and (H) after enucleation.</p>", "links"=>[], "tags"=>["mii", "chromosomes", "oocytes", "fluorescence", "halogen"], "article_id"=>356107, "categories"=>["Physics", "Biotechnology", "Science Policy", "Genetics", "Developmental Biology"], "users"=>["Kazuo Yamagata", "Daisaku Iwamoto", "Yukari Terashita", "Chong Li", "Sayaka Wakayama", "Yoko Hayashi-Takanaka", "Hiroshi Kimura", "Kazuhiro Saeki", "Teruhiko Wakayama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031638.g004", "stats"=>{"downloads"=>4, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Enucleation_of_MII_chromosomes_from_oocytes_with_fluorescence_observation_using_halogen_lamp_/356107", "title"=>"Enucleation of MII chromosomes from oocytes with fluorescence observation using halogen lamp.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-08 01:41:47"}
• {"files"=>["https://ndownloader.figshare.com/files/685734"], "description"=>"<p>Toxicity of Hoechst and mercury lamp for mouse embryo development.</p>", "links"=>[], "tags"=>["hoechst", "mercury", "embryo"], "article_id"=>356211, "categories"=>["Physics", "Biotechnology", "Science Policy", "Genetics", "Developmental Biology"], "users"=>["Kazuo Yamagata", "Daisaku Iwamoto", "Yukari Terashita", "Chong Li", "Sayaka Wakayama", "Yoko Hayashi-Takanaka", "Hiroshi Kimura", "Kazuhiro Saeki", "Teruhiko Wakayama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031638.t001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Toxicity_of_Hoechst_and_mercury_lamp_for_mouse_embryo_development_/356211", "title"=>"Toxicity of Hoechst and mercury lamp for mouse embryo development.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-02-08 01:43:31"}
• {"files"=>["https://ndownloader.figshare.com/files/685202"], "description"=>"<p>Fixed mouse blastocysts were treated with anti-Cdx2- and anti-Oct3/4-antibodies and stained with Alexa Fluor 488 (green) and 548 (red), respectively, then observed using a mercury vapor lamp (A–C) or an inverted microscope with a halogen lamp (D–F). Although the images produced by halogen light illumination were slightly weaker than images produced with the mercury vapor lamp, they could substitute for those seen using a traditional fluorescence microscope. Similar specimens were observed using an upright microscope with halogen lamp (G–L). These images were taken using an LCPlanF1 objective lens (×20; bar = 100 µm). (J) Bright field illumination. (K, L) A different focal plane of the same embryo shown in (I). (J) Inner cell mass (ICM) cells appear as Oct3/4-positive cells. (K, L) Merged images. (J–L) Observed using an LCPlanF1 objective lens (×40; bar = 50 µm).</p>", "links"=>[], "tags"=>["fluorescent", "dyes", "halogen"], "article_id"=>355681, "categories"=>["Physics", "Biotechnology", "Science Policy", "Genetics", "Developmental Biology"], "users"=>["Kazuo Yamagata", "Daisaku Iwamoto", "Yukari Terashita", "Chong Li", "Sayaka Wakayama", "Yoko Hayashi-Takanaka", "Hiroshi Kimura", "Kazuhiro Saeki", "Teruhiko Wakayama"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031638.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Observation_of_fluorescent_dyes_using_a_halogen_lamp_/355681", "title"=>"Observation of fluorescent dyes using a halogen lamp.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-08 01:34:41"}

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Relative Metric

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F1000Prime