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{"title"=>"Intrinsic indicator of photodamage during label-free multiphoton microscopy of cells and tissues", "type"=>"journal", "authors"=>[{"first_name"=>"Roberta", "last_name"=>"Galli", "scopus_author_id"=>"55218793500"}, {"first_name"=>"Ortrud", "last_name"=>"Uckermann", "scopus_author_id"=>"6603124234"}, {"first_name"=>"Elisabeth F.", "last_name"=>"Andresen", "scopus_author_id"=>"56400497000"}, {"first_name"=>"Kathrin D.", "last_name"=>"Geiger", "scopus_author_id"=>"7005822850"}, {"first_name"=>"Edmund", "last_name"=>"Koch", "scopus_author_id"=>"7201389471"}, {"first_name"=>"Gabriele", "last_name"=>"Schackert", "scopus_author_id"=>"7006398216"}, {"first_name"=>"Gerald", "last_name"=>"Steiner", "scopus_author_id"=>"56468197900"}, {"first_name"=>"Matthias", "last_name"=>"Kirsch", "scopus_author_id"=>"7102663025"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"doi"=>"10.1371/journal.pone.0110295", "sgr"=>"84908428906", "issn"=>"19326203", "pui"=>"600290405", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"25343251", "scopus"=>"2-s2.0-84908428906"}, "id"=>"4179a8f2-8bd7-3cbc-8521-bc0ae10b1bc2", "abstract"=>"Multiphoton imaging has evolved as an indispensable tool in cell biology and holds prospects for clinical applications. When addressing endogenous signals such as coherent anti-Stokes Raman scattering (CARS) or second harmonic generation, it requires intense laser irradiation that may cause photodamage. We report that increasing endogenous fluorescence signal upon multiphoton imaging constitutes a marker of photodamage. The effect was studied on mouse brain in vivo and ex vivo, on ex vivo human brain tissue samples, as well as on glioblastoma cells in vitro, demonstrating that this phenomenon is common to a variety of different systems, both ex vivo and in vivo. CARS microscopy and vibrational spectroscopy were used to analyze the photodamage. The development of a standard easy-to-use model that employs rehydrated cryosections allowed the characterization of the irradiation-induced fluorescence and related it to nonlinear photodamage. In conclusion, the monitoring of endogenous two-photon excited fluorescence during label-free multiphoton microscopy enables to estimate damage thresholds ex vivo as well as detect photodamage during in vivo experiments.", "link"=>"http://www.mendeley.com/research/intrinsic-indicator-photodamage-during-labelfree-multiphoton-microscopy-cells-tissues", "reader_count"=>40, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>13, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>3, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>2, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>13, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>3, "Student > Master"=>5, "Other"=>2, "Student > Bachelor"=>2, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>3, "Unspecified"=>3, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>10, "Medicine and Dentistry"=>4, "Neuroscience"=>1, "Business, Management and Accounting"=>1, "Physics and Astronomy"=>11, "Chemistry"=>3}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>3}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>3}, "Physics and Astronomy"=>{"Physics and Astronomy"=>11}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>10}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>3}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"Finland"=>1, "Israel"=>1, "Switzerland"=>1}, "group_count"=>2}

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/4350184", "https://ndownloader.figshare.com/files/4350190", "https://ndownloader.figshare.com/files/4350199", "https://ndownloader.figshare.com/files/4350202", "https://ndownloader.figshare.com/files/4350208", "https://ndownloader.figshare.com/files/4350214", "https://ndownloader.figshare.com/files/4350220", "https://ndownloader.figshare.com/files/4350226"], "description"=>"<div><p>Multiphoton imaging has evolved as an indispensable tool in cell biology and holds prospects for clinical applications. When addressing endogenous signals such as coherent anti-Stokes Raman scattering (CARS) or second harmonic generation, it requires intense laser irradiation that may cause photodamage. We report that increasing endogenous fluorescence signal upon multiphoton imaging constitutes a marker of photodamage. The effect was studied on mouse brain in vivo and ex vivo, on ex vivo human brain tissue samples, as well as on glioblastoma cells in vitro, demonstrating that this phenomenon is common to a variety of different systems, both ex vivo and in vivo. CARS microscopy and vibrational spectroscopy were used to analyze the photodamage. The development of a standard easy-to-use model that employs rehydrated cryosections allowed the characterization of the irradiation-induced fluorescence and related it to nonlinear photodamage. In conclusion, the monitoring of endogenous two-photon excited fluorescence during label-free multiphoton microscopy enables to estimate damage thresholds ex vivo as well as detect photodamage during in vivo experiments.</p></div>", "links"=>[], "tags"=>["Intrinsic Indicator", "multiphoton imaging", "fluorescence signal", "cell biology", "nonlinear photodamage", "vivo experiments", "rehydrated cryosections", "mouse brain", "laser irradiation", "Tissues Multiphoton imaging", "glioblastoma cells", "vibrational spectroscopy", "estimate damage thresholds", "brain tissue samples", "CARS microscopy"], "article_id"=>1220480, "categories"=>["Biochemistry", "Space Science", "Medicine", "Cell Biology", "Molecular Biology", "Physiology", "Biotechnology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Cancer"], "users"=>["Roberta Galli", "Ortrud Uckermann", "Elisabeth F. Andresen", "Kathrin D. Geiger", "Edmund Koch", "Gabriele Schackert", "Gerald Steiner", "Matthias Kirsch"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0110295.s001", "https://dx.doi.org/10.1371/journal.pone.0110295.s002", "https://dx.doi.org/10.1371/journal.pone.0110295.s003", "https://dx.doi.org/10.1371/journal.pone.0110295.s004", "https://dx.doi.org/10.1371/journal.pone.0110295.s005", "https://dx.doi.org/10.1371/journal.pone.0110295.s006", "https://dx.doi.org/10.1371/journal.pone.0110295.s007", "https://dx.doi.org/10.1371/journal.pone.0110295.s008"], "stats"=>{"downloads"=>0, "page_views"=>24, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Intrinsic_Indicator_of_Photodamage_during_Label_Free_Multiphoton_Microscopy_of_Cells_and_Tissues_/1220480", "title"=>"Intrinsic Indicator of Photodamage during Label-Free Multiphoton Microscopy of Cells and Tissues", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-24 17:27:35"}
• {"files"=>["https://ndownloader.figshare.com/files/4350250"], "description"=>"<p><b>A</b>: Multiphoton image of cortical tissue after 240 scans in the area indicated by the box and TPEF images showing the irradiated area at scan n. 1, 94, 168 and 240; the arrow indicates the very first increase of TPEF at scan 110. <b>B</b>: Cerebellar tissue after 224 scans in the area indicated by the box and TPEF images showing the irradiated area at scan n. 1, 108, 162 and 224; the arrow indicates the very first increase of TPEF at scan n. 108. In both cases H&E staining of a consecutive section is provided as reference for tissue structures. <b>C</b>: Magnified image of the region indicated by the dashed box in B: TPEF (green) and CARS (red) are shown separately and merged. Arrows indicate spots of strong TPEF that colocalize with decreased CARS signal intensity. All experiments were performed with a total laser power of ∼52 mW in the sample.</p>", "links"=>[], "tags"=>["Intrinsic Indicator", "multiphoton imaging", "fluorescence signal", "cell biology", "nonlinear photodamage", "vivo experiments", "rehydrated cryosections", "mouse brain", "laser irradiation", "Tissues Multiphoton imaging", "glioblastoma cells", "vibrational spectroscopy", "estimate damage thresholds", "brain tissue samples", "CARS microscopy"], "article_id"=>1217723, "categories"=>["Biochemistry", "Space Science", "Medicine", "Cell Biology", "Molecular Biology", "Physiology", "Biotechnology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Cancer"], "users"=>["Roberta Galli", "Ortrud Uckermann", "Elisabeth F. Andresen", "Kathrin D. Geiger", "Edmund Koch", "Gabriele Schackert", "Gerald Steiner", "Matthias Kirsch"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0110295.g003"], "stats"=>{"downloads"=>0, "page_views"=>50, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Endogenous_fluorescence_induced_by_multiphoton_microscopy_on_rehydrated_cryosections_of_mouse_brain_/1217723", "title"=>"Endogenous fluorescence induced by multiphoton microscopy on rehydrated cryosections of mouse brain.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-24 17:27:35"}
• {"files"=>["https://ndownloader.figshare.com/files/4350262"], "description"=>"<p>The repetitive irradiation was performed in the central part of the images. CARS (red), TPEF (green) and SHG (blue) signals were acquired. The total number of scans used to produce the damage is also reported. The TPEF images of the first and last scan are shown for comparison. All irradiations were performed using the same experimental settings and therefore the scan numbers can be compared. Histological H&E staining of consecutive sections is reported as reference to visualize the tissue structures. All experiments were performed with a total laser power of ∼52 mW in the sample.</p>", "links"=>[], "tags"=>["Intrinsic Indicator", "multiphoton imaging", "fluorescence signal", "cell biology", "nonlinear photodamage", "vivo experiments", "rehydrated cryosections", "mouse brain", "laser irradiation", "Tissues Multiphoton imaging", "glioblastoma cells", "vibrational spectroscopy", "estimate damage thresholds", "brain tissue samples", "CARS microscopy"], "article_id"=>1217735, "categories"=>["Biochemistry", "Space Science", "Medicine", "Cell Biology", "Molecular Biology", "Physiology", "Biotechnology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Cancer"], "users"=>["Roberta Galli", "Ortrud Uckermann", "Elisabeth F. Andresen", "Kathrin D. Geiger", "Edmund Koch", "Gabriele Schackert", "Gerald Steiner", "Matthias Kirsch"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0110295.g005"], "stats"=>{"downloads"=>1, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Photodamage_on_rehydrated_cryosections_of_different_mouse_tissue_types_/1217735", "title"=>"Photodamage on rehydrated cryosections of different mouse tissue types.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-24 17:27:35"}
• {"files"=>["https://ndownloader.figshare.com/files/4350265"], "description"=>"<p><b>A</b>: Raman and FT-IR spectra of an irradiated mouse brain cortex showing increased endogenous fluorescence. The spectra were obtained by subtraction of the not irradiated tissue spectrum from the irradiated one. The most pronounced positive and negative bands are labeled and discussed in the text. <b>B</b>: Transmission electron microscopy of an irradiated sample of human brain tissue (white matter), showing partially destroyed myelin sheaths (*), a partly damaged nucleus (+) and preserved axons (#).</p>", "links"=>[], "tags"=>["Intrinsic Indicator", "multiphoton imaging", "fluorescence signal", "cell biology", "nonlinear photodamage", "vivo experiments", "rehydrated cryosections", "mouse brain", "laser irradiation", "Tissues Multiphoton imaging", "glioblastoma cells", "vibrational spectroscopy", "estimate damage thresholds", "brain tissue samples", "CARS microscopy"], "article_id"=>1217739, "categories"=>["Biochemistry", "Space Science", "Medicine", "Cell Biology", "Molecular Biology", "Physiology", "Biotechnology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Cancer"], "users"=>["Roberta Galli", "Ortrud Uckermann", "Elisabeth F. Andresen", "Kathrin D. Geiger", "Edmund Koch", "Gabriele Schackert", "Gerald Steiner", "Matthias Kirsch"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0110295.g006"], "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_tissue_alterations_connected_to_the_irradiation_induced_fluorescence_/1217739", "title"=>"Characterization of tissue alterations connected to the irradiation-induced fluorescence.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-24 17:27:35"}
• {"files"=>["https://ndownloader.figshare.com/files/4350256"], "description"=>"<p>The rate was obtained as inverse of the number of scans required to attain an increase of TPEF of 25% above baseline. The best fit line has a slope of 3.17±0.24 on a logarithmic scale.</p>", "links"=>[], "tags"=>["Intrinsic Indicator", "multiphoton imaging", "fluorescence signal", "cell biology", "nonlinear photodamage", "vivo experiments", "rehydrated cryosections", "mouse brain", "laser irradiation", "Tissues Multiphoton imaging", "glioblastoma cells", "vibrational spectroscopy", "estimate damage thresholds", "brain tissue samples", "CARS microscopy"], "article_id"=>1217724, "categories"=>["Biochemistry", "Space Science", "Medicine", "Cell Biology", "Molecular Biology", "Physiology", "Biotechnology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Cancer"], "users"=>["Roberta Galli", "Ortrud Uckermann", "Elisabeth F. Andresen", "Kathrin D. Geiger", "Edmund Koch", "Gabriele Schackert", "Gerald Steiner", "Matthias Kirsch"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0110295.g004"], "stats"=>{"downloads"=>1, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dependence_of_the_TPEF_increase_rate_on_laser_power_/1217724", "title"=>"Dependence of the TPEF increase rate on laser power.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-24 17:27:35"}
• {"files"=>["https://ndownloader.figshare.com/files/4350244"], "description"=>"<p><b>A</b>: Bright field (BF) and TPEF image of the area in the box in the BF image acquired on living primary human glioblastoma cells in culture, and overly of BF and TPEF images. <b>B</b>: effects of a serial irradiation of 500 scans at laser power of ∼52 mW in the area indicated by the box, leading to increase of TPEF in the cell cytoplasm. C: Label-free multiphoton image and fluorescence image (acquired using a mercury lamp for excitation) of mouse brain cortex; the brain surface is slightly convex, so that the image cuts through different tissue layers: CARS shows the cortical tissue with a blood vessel, while SHG shows the inner meningeal layer on top of the cortex. D: Label-free multiphoton image and fluorescence image acquired in the same region shown in C after 544 scans performed in vivo in the area indicated by the dotted line box; the arrows indicate the localized increase of fluorescence that matches the increase of TPEF.</p>", "links"=>[], "tags"=>["Intrinsic Indicator", "multiphoton imaging", "fluorescence signal", "cell biology", "nonlinear photodamage", "vivo experiments", "rehydrated cryosections", "mouse brain", "laser irradiation", "Tissues Multiphoton imaging", "glioblastoma cells", "vibrational spectroscopy", "estimate damage thresholds", "brain tissue samples", "CARS microscopy"], "article_id"=>1217721, "categories"=>["Biochemistry", "Space Science", "Medicine", "Cell Biology", "Molecular Biology", "Physiology", "Biotechnology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Cancer"], "users"=>["Roberta Galli", "Ortrud Uckermann", "Elisabeth F. Andresen", "Kathrin D. Geiger", "Edmund Koch", "Gabriele Schackert", "Gerald Steiner", "Matthias Kirsch"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0110295.g002"], "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_photodamage_on_living_organisms_during_label_free_multiphoton_imaging_/1217721", "title"=>"Effects of photodamage on living organisms during label-free multiphoton imaging.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-24 17:27:35"}
• {"files"=>["https://ndownloader.figshare.com/files/4350238"], "description"=>"<p><b>A</b>: Single high quality image of a Purkinje cell acquired using CARS (red) and TPEF (green) at laser excitation power ∼52 mW. <b>B</b>: Further serial scanning of the area indicated by the dotted-line box in A resulted in a gradual increase in TPEF. The cell body is shown after 30, 164 and 272 scans. The photodamage-induced TPEF developed first in the tissue around the cell body and then also inside the cell cytoplasm. <b>C</b>: TPEF difference images (corresponding to the images in B) obtained subtracting the TPEF image of the first scan from the TPEF images at 30, 164 and 272 scans respectively; they underline the photodamage-induced TPEF around the cell (see scan 164) and in the cell cytoplasm (see scan 272).</p>", "links"=>[], "tags"=>["Intrinsic Indicator", "multiphoton imaging", "fluorescence signal", "cell biology", "nonlinear photodamage", "vivo experiments", "rehydrated cryosections", "mouse brain", "laser irradiation", "Tissues Multiphoton imaging", "glioblastoma cells", "vibrational spectroscopy", "estimate damage thresholds", "brain tissue samples", "CARS microscopy"], "article_id"=>1217718, "categories"=>["Biochemistry", "Space Science", "Medicine", "Cell Biology", "Molecular Biology", "Physiology", "Biotechnology", "Biological Sciences not elsewhere classified", "Developmental Biology", "Cancer"], "users"=>["Roberta Galli", "Ortrud Uckermann", "Elisabeth F. Andresen", "Kathrin D. Geiger", "Edmund Koch", "Gabriele Schackert", "Gerald Steiner", "Matthias Kirsch"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0110295.g001"], "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_photodamage_on_ex_vivo_human_cerebellum_tissue_during_label_free_multiphoton_imaging_/1217718", "title"=>"Effects of photodamage on ex vivo human cerebellum tissue during label-free multiphoton imaging.", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-10-24 17:27:35"}

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Relative Metric

{"start_date"=>"2014-01-01T00:00:00Z", "end_date"=>"2014-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Engineering and technology/Equipment", "average_usage"=>[287, 444]}, {"subject_area"=>"/Engineering and technology/Signal processing", "average_usage"=>[276]}]}

F1000Prime