Autofluorescence imaging of human RPE cell granules using structured illumination microscopy
Background/aimsTo characterise single autofluorescent (AF) granules in human retinal pigment epithelium (RPE) cells using structured illumination microscopy (SIM).MethodsMorphological characteristics and autofluorescence behaviour of lipofuscin (LF) and melanolipofuscin (MLF) granules of macular RPE...
Gespeichert in:
| Veröffentlicht in: | British journal of ophthalmology 2012-08, Vol.96 (8), p.1141-1144 |
|---|---|
| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1144 |
|---|---|
| container_issue | 8 |
| container_start_page | 1141 |
| container_title | British journal of ophthalmology |
| container_volume | 96 |
| creator | Ach, Thomas Best, Gerrit Rossberger, Sabrina Heintzmann, Rainer Cremer, Christoph Dithmar, Stefan |
| description | Background/aimsTo characterise single autofluorescent (AF) granules in human retinal pigment epithelium (RPE) cells using structured illumination microscopy (SIM).MethodsMorphological characteristics and autofluorescence behaviour of lipofuscin (LF) and melanolipofuscin (MLF) granules of macular RPE cells (66-year-old donor) were examined with SIM using three different laser light excitation wavelengths (488, 568 and 647 nm). High-resolution images were reconstructed and exported to Matlab R2009a (The Mathworks Inc, Natick, MA, USA) to determine accurate size and emission intensities of LF and MLF granules.ResultsSIM doubles lateral resolution compared with conventionally used wide-field microscopy and allows visualisation of intracellular structures down to 110 nm lateral resolution. AF patterns were examined in 133 LF and 27 MLF granules. LF granules (968±220 nm) were significantly smaller in diameter than MLF granules (1097±110 nm; p |
| doi_str_mv | 10.1136/bjophthalmol-2012-301547 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1028020225</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>4012521951</sourcerecordid><originalsourceid>FETCH-LOGICAL-b521t-4c910f46772fe604cad04c10d965c082944a862e73758b4d9f9376de552ad353</originalsourceid><addsrcrecordid>eNqNkVFrHCEUhaU0NNu0f6EIodCXadVx1HlMlqQtXZJQlpCHgjiOs-vWGbc6QvLv4zDbtOQpL4r63XuP5wAAMfqMccm-NDu_345b5XrvCoIwKUqEK8pfgQWmTOQrXr8GC4QQLzBm-Bi8jXGXj4Rh_gYcE8IZooIvwK-zNPrOJR9M1GbQBtpebeywgb6D29SrAf68uYDaOAc3QQ3JmQhTnIA4hqTHFEwLrXOpt4MarR9gb3XwUfv9wztw1CkXzfvDfgLWlxfr5bdidf31-_JsVTQVwWNBdY1RRxnnpDNZl1ZtXjBqa1ZpJEhNqRKMGF7ySjS0rbu65Kw1VUVUW1blCfg0t90H_yeZOMrexkmxGoxPUWJEBCKIkAk9fYbufApDFicx56Km2R-cKTFT00diMJ3ch2xLeMit5BSA_D8AOQUg5wBy6YfDgNT0pn0q_Ot4Bj4eABW1cl32VNv4j2O4JqJimStmzsbR3D-9q_BbsskIeXW7lOd8fcd_rK4kyXw5802_e7ncR2PvsgU</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1778946171</pqid></control><display><type>article</type><title>Autofluorescence imaging of human RPE cell granules using structured illumination microscopy</title><source>MEDLINE</source><source>BMJ Journals - NESLi2</source><source>PubMed Central</source><creator>Ach, Thomas ; Best, Gerrit ; Rossberger, Sabrina ; Heintzmann, Rainer ; Cremer, Christoph ; Dithmar, Stefan</creator><creatorcontrib>Ach, Thomas ; Best, Gerrit ; Rossberger, Sabrina ; Heintzmann, Rainer ; Cremer, Christoph ; Dithmar, Stefan</creatorcontrib><description>Background/aimsTo characterise single autofluorescent (AF) granules in human retinal pigment epithelium (RPE) cells using structured illumination microscopy (SIM).MethodsMorphological characteristics and autofluorescence behaviour of lipofuscin (LF) and melanolipofuscin (MLF) granules of macular RPE cells (66-year-old donor) were examined with SIM using three different laser light excitation wavelengths (488, 568 and 647 nm). High-resolution images were reconstructed and exported to Matlab R2009a (The Mathworks Inc, Natick, MA, USA) to determine accurate size and emission intensities of LF and MLF granules.ResultsSIM doubles lateral resolution compared with conventionally used wide-field microscopy and allows visualisation of intracellular structures down to 110 nm lateral resolution. AF patterns were examined in 133 LF and 27 MLF granules. LF granules (968±220 nm) were significantly smaller in diameter than MLF granules (1097±110 nm; p<0.001). LF granules showed an inhomogeneous intragranular pattern, and the average intensity negatively correlated with the size of these granules when excited at 647 nm. The autofluorescence of MLF granules was more homogeneous, but shifted towards higher excitation wavelengths in the centre of the granules.ConclusionSIM is a useful tool for examining AF signals within single LF and MLF granules in RPE cells. This allows new insights into RPE autofluorescence patterns.</description><identifier>ISSN: 0007-1161</identifier><identifier>EISSN: 1468-2079</identifier><identifier>DOI: 10.1136/bjophthalmol-2012-301547</identifier><identifier>PMID: 22760487</identifier><identifier>CODEN: BJOPAL</identifier><language>eng</language><publisher>BMA House, Tavistock Square, London, WC1H 9JR: BMJ Publishing Group Ltd</publisher><subject>Aged ; Autofluorescence ; Biological and medical sciences ; Cytoplasmic Granules - metabolism ; Fluorescence ; Humans ; Lasers ; lipofuscin ; Lipofuscin - metabolism ; Macular degeneration ; Male ; Medical sciences ; melanolipofuscin ; Microscopy ; Microscopy, Fluorescence ; Miscellaneous ; Morphology ; Ophthalmology ; Pathogenesis ; Retinal Pigment Epithelium - cytology ; Retinal Pigment Epithelium - metabolism ; Software ; structured illumination ; Tissue Donors</subject><ispartof>British journal of ophthalmology, 2012-08, Vol.96 (8), p.1141-1144</ispartof><rights>2012, Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.</rights><rights>2015 INIST-CNRS</rights><rights>Copyright: 2012 (c) 2012, Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b521t-4c910f46772fe604cad04c10d965c082944a862e73758b4d9f9376de552ad353</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttp://bjo.bmj.com/content/96/8/1141.full.pdf$$EPDF$$P50$$Gbmj$$H</linktopdf><linktohtml>$$Uhttp://bjo.bmj.com/content/96/8/1141.full$$EHTML$$P50$$Gbmj$$H</linktohtml><link.rule.ids>114,115,314,777,781,3183,23552,27905,27906,77349,77380</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=26192856$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22760487$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ach, Thomas</creatorcontrib><creatorcontrib>Best, Gerrit</creatorcontrib><creatorcontrib>Rossberger, Sabrina</creatorcontrib><creatorcontrib>Heintzmann, Rainer</creatorcontrib><creatorcontrib>Cremer, Christoph</creatorcontrib><creatorcontrib>Dithmar, Stefan</creatorcontrib><title>Autofluorescence imaging of human RPE cell granules using structured illumination microscopy</title><title>British journal of ophthalmology</title><addtitle>Br J Ophthalmol</addtitle><description>Background/aimsTo characterise single autofluorescent (AF) granules in human retinal pigment epithelium (RPE) cells using structured illumination microscopy (SIM).MethodsMorphological characteristics and autofluorescence behaviour of lipofuscin (LF) and melanolipofuscin (MLF) granules of macular RPE cells (66-year-old donor) were examined with SIM using three different laser light excitation wavelengths (488, 568 and 647 nm). High-resolution images were reconstructed and exported to Matlab R2009a (The Mathworks Inc, Natick, MA, USA) to determine accurate size and emission intensities of LF and MLF granules.ResultsSIM doubles lateral resolution compared with conventionally used wide-field microscopy and allows visualisation of intracellular structures down to 110 nm lateral resolution. AF patterns were examined in 133 LF and 27 MLF granules. LF granules (968±220 nm) were significantly smaller in diameter than MLF granules (1097±110 nm; p<0.001). LF granules showed an inhomogeneous intragranular pattern, and the average intensity negatively correlated with the size of these granules when excited at 647 nm. The autofluorescence of MLF granules was more homogeneous, but shifted towards higher excitation wavelengths in the centre of the granules.ConclusionSIM is a useful tool for examining AF signals within single LF and MLF granules in RPE cells. This allows new insights into RPE autofluorescence patterns.</description><subject>Aged</subject><subject>Autofluorescence</subject><subject>Biological and medical sciences</subject><subject>Cytoplasmic Granules - metabolism</subject><subject>Fluorescence</subject><subject>Humans</subject><subject>Lasers</subject><subject>lipofuscin</subject><subject>Lipofuscin - metabolism</subject><subject>Macular degeneration</subject><subject>Male</subject><subject>Medical sciences</subject><subject>melanolipofuscin</subject><subject>Microscopy</subject><subject>Microscopy, Fluorescence</subject><subject>Miscellaneous</subject><subject>Morphology</subject><subject>Ophthalmology</subject><subject>Pathogenesis</subject><subject>Retinal Pigment Epithelium - cytology</subject><subject>Retinal Pigment Epithelium - metabolism</subject><subject>Software</subject><subject>structured illumination</subject><subject>Tissue Donors</subject><issn>0007-1161</issn><issn>1468-2079</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNqNkVFrHCEUhaU0NNu0f6EIodCXadVx1HlMlqQtXZJQlpCHgjiOs-vWGbc6QvLv4zDbtOQpL4r63XuP5wAAMfqMccm-NDu_345b5XrvCoIwKUqEK8pfgQWmTOQrXr8GC4QQLzBm-Bi8jXGXj4Rh_gYcE8IZooIvwK-zNPrOJR9M1GbQBtpebeywgb6D29SrAf68uYDaOAc3QQ3JmQhTnIA4hqTHFEwLrXOpt4MarR9gb3XwUfv9wztw1CkXzfvDfgLWlxfr5bdidf31-_JsVTQVwWNBdY1RRxnnpDNZl1ZtXjBqa1ZpJEhNqRKMGF7ySjS0rbu65Kw1VUVUW1blCfg0t90H_yeZOMrexkmxGoxPUWJEBCKIkAk9fYbufApDFicx56Km2R-cKTFT00diMJ3ch2xLeMit5BSA_D8AOQUg5wBy6YfDgNT0pn0q_Ot4Bj4eABW1cl32VNv4j2O4JqJimStmzsbR3D-9q_BbsskIeXW7lOd8fcd_rK4kyXw5802_e7ncR2PvsgU</recordid><startdate>20120801</startdate><enddate>20120801</enddate><creator>Ach, Thomas</creator><creator>Best, Gerrit</creator><creator>Rossberger, Sabrina</creator><creator>Heintzmann, Rainer</creator><creator>Cremer, Christoph</creator><creator>Dithmar, Stefan</creator><general>BMJ Publishing Group Ltd</general><general>BMJ Publishing Group</general><general>BMJ Publishing Group LTD</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>BTHHO</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope></search><sort><creationdate>20120801</creationdate><title>Autofluorescence imaging of human RPE cell granules using structured illumination microscopy</title><author>Ach, Thomas ; Best, Gerrit ; Rossberger, Sabrina ; Heintzmann, Rainer ; Cremer, Christoph ; Dithmar, Stefan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b521t-4c910f46772fe604cad04c10d965c082944a862e73758b4d9f9376de552ad353</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Aged</topic><topic>Autofluorescence</topic><topic>Biological and medical sciences</topic><topic>Cytoplasmic Granules - metabolism</topic><topic>Fluorescence</topic><topic>Humans</topic><topic>Lasers</topic><topic>lipofuscin</topic><topic>Lipofuscin - metabolism</topic><topic>Macular degeneration</topic><topic>Male</topic><topic>Medical sciences</topic><topic>melanolipofuscin</topic><topic>Microscopy</topic><topic>Microscopy, Fluorescence</topic><topic>Miscellaneous</topic><topic>Morphology</topic><topic>Ophthalmology</topic><topic>Pathogenesis</topic><topic>Retinal Pigment Epithelium - cytology</topic><topic>Retinal Pigment Epithelium - metabolism</topic><topic>Software</topic><topic>structured illumination</topic><topic>Tissue Donors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ach, Thomas</creatorcontrib><creatorcontrib>Best, Gerrit</creatorcontrib><creatorcontrib>Rossberger, Sabrina</creatorcontrib><creatorcontrib>Heintzmann, Rainer</creatorcontrib><creatorcontrib>Cremer, Christoph</creatorcontrib><creatorcontrib>Dithmar, Stefan</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>BMJ Journals</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><jtitle>British journal of ophthalmology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ach, Thomas</au><au>Best, Gerrit</au><au>Rossberger, Sabrina</au><au>Heintzmann, Rainer</au><au>Cremer, Christoph</au><au>Dithmar, Stefan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Autofluorescence imaging of human RPE cell granules using structured illumination microscopy</atitle><jtitle>British journal of ophthalmology</jtitle><addtitle>Br J Ophthalmol</addtitle><date>2012-08-01</date><risdate>2012</risdate><volume>96</volume><issue>8</issue><spage>1141</spage><epage>1144</epage><pages>1141-1144</pages><issn>0007-1161</issn><eissn>1468-2079</eissn><coden>BJOPAL</coden><abstract>Background/aimsTo characterise single autofluorescent (AF) granules in human retinal pigment epithelium (RPE) cells using structured illumination microscopy (SIM).MethodsMorphological characteristics and autofluorescence behaviour of lipofuscin (LF) and melanolipofuscin (MLF) granules of macular RPE cells (66-year-old donor) were examined with SIM using three different laser light excitation wavelengths (488, 568 and 647 nm). High-resolution images were reconstructed and exported to Matlab R2009a (The Mathworks Inc, Natick, MA, USA) to determine accurate size and emission intensities of LF and MLF granules.ResultsSIM doubles lateral resolution compared with conventionally used wide-field microscopy and allows visualisation of intracellular structures down to 110 nm lateral resolution. AF patterns were examined in 133 LF and 27 MLF granules. LF granules (968±220 nm) were significantly smaller in diameter than MLF granules (1097±110 nm; p<0.001). LF granules showed an inhomogeneous intragranular pattern, and the average intensity negatively correlated with the size of these granules when excited at 647 nm. The autofluorescence of MLF granules was more homogeneous, but shifted towards higher excitation wavelengths in the centre of the granules.ConclusionSIM is a useful tool for examining AF signals within single LF and MLF granules in RPE cells. This allows new insights into RPE autofluorescence patterns.</abstract><cop>BMA House, Tavistock Square, London, WC1H 9JR</cop><pub>BMJ Publishing Group Ltd</pub><pmid>22760487</pmid><doi>10.1136/bjophthalmol-2012-301547</doi><tpages>4</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0007-1161 |
| ispartof | British journal of ophthalmology, 2012-08, Vol.96 (8), p.1141-1144 |
| issn | 0007-1161 1468-2079 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1028020225 |
| source | MEDLINE; BMJ Journals - NESLi2; PubMed Central |
| subjects | Aged Autofluorescence Biological and medical sciences Cytoplasmic Granules - metabolism Fluorescence Humans Lasers lipofuscin Lipofuscin - metabolism Macular degeneration Male Medical sciences melanolipofuscin Microscopy Microscopy, Fluorescence Miscellaneous Morphology Ophthalmology Pathogenesis Retinal Pigment Epithelium - cytology Retinal Pigment Epithelium - metabolism Software structured illumination Tissue Donors |
| title | Autofluorescence imaging of human RPE cell granules using structured illumination microscopy |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-20T10%3A47%3A48IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Autofluorescence%20imaging%20of%20human%20RPE%20cell%20granules%20using%20structured%20illumination%20microscopy&rft.jtitle=British%20journal%20of%20ophthalmology&rft.au=Ach,%20Thomas&rft.date=2012-08-01&rft.volume=96&rft.issue=8&rft.spage=1141&rft.epage=1144&rft.pages=1141-1144&rft.issn=0007-1161&rft.eissn=1468-2079&rft.coden=BJOPAL&rft_id=info:doi/10.1136/bjophthalmol-2012-301547&rft_dat=%3Cproquest_cross%3E4012521951%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1778946171&rft_id=info:pmid/22760487&rfr_iscdi=true |